Systems and methods for manufacturing biostructures

Abstract

Described herein are novel construction material compositions, systems, and methods for manufacturing the same. Biocement technologies that monitor and adjust environmental and process controls during the manufacturing of biocement products are described. In some cases, during manufacturing of a construction material various manufacturing parameters including feed rate, feed pressure, feed molarity, feed rate profile, feed pH level, temperature, humidity, the percentage of fine limestone within an aggregate mix, the thickness of a pre-feed crust layer may be set and / or adjusted over time to reduce manufacturing time and to improve the structural integrity of a construction material.

Description

SYSTEMS AND METHODS FOR MANUFACTURING BIOSTRUCTURESBACKGROUND

[0001] Biocement technologies offer cost effective high-strength building materials, structural materials, and concretes which can have a substantially reduced carbon emission footprint compared to traditional building materials and concretes. Accordingly, the ability to manufacture biostructures that include biocement compositions may encourage the replacement of traditional building materials and concretes with biocement products.BRIEF SUMMARY

[0002] Biocement technologies utilize biology (e.g., enzymes or microorganisms, such as mineralizing enzymes or microorganisms) to produce, and / or improve the mechanical and structural properties of construction materials. In some embodiments, the biocement technologies utilize enzymes or microorganisms to form mineral binders that act as binding agents within a construction material. In one example, through microbiologically induced carbonate precipitation (MICP) (also known as enzyme-induced carbonate precipitation (EICP)), the enzymes or microorganisms react with chemical components induce the formation of the mineral binders in the form of compounds that act as binding agents within the construction material. A mineral binder may comprise a carbonate (e.g., dolomite or calcium carbonate, e.g., calcite, aragonite, or amorphous calcium carbonate), a silicate mineral (e.g., silica or aluminosilicate), a sulfate, or a phosphate (e.g., hydroxyapatite). A mineral binder may be referred to as biocement.

[0003] In some embodiments, demonstrated herein are a series of solid-state enzyme- catalyzed (e.g.. solid state fermentation) technologies, across a range of reagent feeding modalities, that are capable of highly reproduceable and scalable production of biostructures. Further, these technologies, processes, and systems, have been demonstrated to operate across a surprisingly broad, and high range of molarities and osmolarities, a surprisingly broad and low range of pHs, and at surprisingly low moisture contents. Further, these technologies are applicable to a broad range of aggregate blends, including high performance aggregate blends, despite the different, and typically more challenging, masstransfer phenomena of fed cementation processes vs Ordinary Portland Cement (OPC).

[0004] In some embodiments, for certain aggregate blends, droplet-based feeding (e.g., spray feeding) can cause surface damage (e.g., by erosion of the metastable shape) during a feeding process, leading to a rougher surface that is more difficult to polish. It has been discovered that this issue may be overcome by modulating average drop size and terminal velocity (e.g., by modulating pump pressure, nozzle height, nozzle aperture size and / or shape, spray angle, or a combination thereof) such that an average droplet impact pressure is between 1 and 2000 Pa (e.g.. between 10 and 1000 Pa, between 1 and 100 Pa, between 1 and 50 Pa, between 10 and 40 Pa, between 1 and 30 Pa, between 1 and 25 Pa, or between 1 and 20 Pa). In some embodiments, the operable pressure range is higher when the shaped structure includes a high surface energy' gelling agent. In some embodiments, the operable pressure range is higher when the shaped structure includes a high angularity aggregate. In some embodiments, the operable pressure range is higher when the shaped structure includes a ratio of a sand fraction to a gravel fraction in the aggregate blend of at least 3:2.

[0005] In some embodiments, the mineral binder within a construction material comprises a metal carbonate that is produced using MICP (also known as EICP), and the construction material comprises one or more aggregates (e.g., combinations of particles of sand, gravel, and crushed stone) that are at least partially bound together via precipitated calcium carbonate crystals. In some cases, the mineral binder within a construction material is combined with a non-biocement mineral binder, such as chemically fused carbonates or chemically fused silicates.

[0006] A biostructure may comprise a structure or construction material that is constructed using one or more biological processes (e.g., using an enzyme or microorganism). In some cases, an enzyme or microorganism is a mineralizing enzyme or microorganism. A constmction material that is constructed using one or more biological processes may include a living construction material if the resulting construction material includes living microorganisms. A construction material that is the result of the application of one or more biological processes may or may not include living biological materials or microorganisms.

[0007] In some cases, biostructure manufacturing parameters including feed volume, feed rate, feed pressure, pre-feed moisture content, feed molarity, feed rate profile, feed pH level, temperature, humidity, the percentage of fines (e.g., fine limestone, pond fines, fine- crushed stone (e.g., granite powder), etc.) within an aggregate mix, and the thickness of a pre-feed crust layer may be set and / or adjusted over time to reduce manufacturing time andto improve the structural integrity of a construction material and / or an intermediate construction material, e.g.. a metastable shaped structure that can withstand a feeding process which results in a final construction material (or hardened intermediate). According to some embodiments, the technical benefits of the methods of manufacturing biostructures disclosed herein include reduced manufacturing time, reduced manufacturing costs, increased construction material throughput, improved freeze-thaw properties, increased flexural strength for the construction material, increased yield (e.g., increased polished yield, e.g., by increasing polish-ability ), and increased compressive strength for the construction material.

[0008] This Summary is provided to introduce a brief description of some aspects of the disclosed technologies in a simplified form that are further described below in the Detailed Description. This Summary is not intended to identify key features or essential features of the claimed subject matter, nor is it intended that this Summary be used to limit the scope of the claimed subject matter. Furthermore, the claimed subject matter is not limited to implementations that solve any or all disadvantages noted in any part of this disclosure.BRIEF DESCRIPTION OF THE DRAWINGS

[0009] Like-numbered elements may refer to common components in the different figures.

[0010] Figure 1 A depicts one embodiment of a biostructure manufacturing system.

[0011] Figure 1 B depicts another embodiment of a biostructure manufacturing system.

[0012] Figure 1C-1E depict various embodiments of a portion of a biostructure manufacturing system that includes a climate-controlled environment.

[0013] Figure I F depicts one embodiment of a portion of a biostructure manufacturing system that includes a feed system that applies a single-pass feed solution to a biostructure.

[0014] Figure 1G depicts one embodiment of a portion of a biostructure manufacturing system that includes a feed system that applies a single-pass feed solution to a biostructure.

[0015] Figures 1H-1L depict various embodiments of a portion of a feed system with one or more spray nozzles for applying a feed solution to a biostructure.

[0016] Figure IM depicts one embodiment of various components of a computing system.

[0017] Figures 1N-1P depict various embodiments of a biostructure manufacturing system.

[0018] Figure 2A depicts one embodiment of a cross-sectional view of a construction material.

[0019] Figure 2B depicts one embodiment of a side view of the construction material shown in Figure 2A.

[0020] Figure 2C depicts one embodiment of aggregate particles within the construction material of Figure 2B prior to compression.

[0021] Figure 2D depicts one embodiment of aggregate particles within the construction material of Figure 2B after compression of the aggregate particles.

[0022] Figure 2E depicts one embodiment of experimental results showing the average compressive strength (in psi) for a construction material over a range of wet green densities (in g / cc) for the construction material prior to the formation of bridging calcium carbonate and / or other binding agents within the construction material.

[0023] Figure 2F depicts one embodiment of a graph showing the percentage of total pore volume vs pore diameter for a construction material.

[0024] Figure 2G depicts one embodiment of experimental results showing the average change in electrical conductivity (or delta EC) for a construction material over a range of average dry green densities (in g / cc) for the construction material.

[0025] Figure 2H depicts one embodiment of a graph showing the total slice porosity over horizontal slice position from a non-preferred surface of a construction material.

[0026] Figure 21 depicts one embodiment of experimental results showing total open pore porosity vs slice number through a construction material.

[0027] Figure 2J depicts one embodiment of a construction material body that includes aggregate particles including aggregate particles that have been bound together using calcium carbonate including calcium carbonate.

[0028] Figures 3A-3B depict a flowchart describing one embodiment of a process for manufacturing a construction material.

[0029] Figures 3C-3D depict a flowchart describing another embodiment of a process for manufacturing a construction material.

[0030] Figure 4A depicts one embodiment of a graphical representation of four feed cycles applied to a construction material.

[0031] Figure 4B depicts one embodiment of a graphical representation of four feed pulses applied to a construction material.

[0032] Figure 4C depicts one embodiment of a set of pulses corresponding to the first feed pulse between times T1 and T2 in Figure 4B.

[0033] Figure 4D depicts one embodiment of a first set of feed pulses

[0034] Figure 4E depicts one embodiment of a first set of feed pulses and a second set of feed pulses.

[0035] Figures 4F-4G depict various embodiments of feed rate profiles.

[0036] Figures 4H-4I depict various embodiments of multiple feed cycles for applying a feed solution to a shaped structure.

[0037] Figure 4J depicts a flowchart describing one embodiment of a process for manufacturing a construction material.

[0038] Figure 4K depicts a flowchart describing another embodiment of a process for manufacturing a construction material.

[0039] Figures 5A-5C depict flowcharts describing various embodiments of a process for manufacturing a construction material.

[0040] Figure 6A depicts a flowchart describing one embodiment of a process for manufacturing a construction material.

[0041] Figure 6B depicts a flowchart describing another embodiment of a process for manufacturing a construction material.

[0042] Figure 6C depicts a flowchart describing an alternative embodiment of a process for manufacturing a construction material.

[0043] Figure 7A shows the scanning electron microscopy results of a series of experiments in which 125 cc cubes were inoculated with different initial cell counts (where lx = approximately 5 xlO8CFU per ml).

[0044] Figures 7B-7D show the results of an experiment in which the pH was modulated at two different cementation reagent molarities (CaCh : Urea = 1: 1). The form factor was125 cc cubes. The feeding was by a drip-based, single pass feed method. The fine component of the aggregate mixture included pond fines.

[0045] Figures 7E-7F show the results of an experiment in which 125 cc cubes with 2% limestone instead of pond fines. These cubes were topically inoculated with 45 mL of an inoculation solution with a cell concentration of 4.5x109cells per mL. The cubes were fed 16 times, with 2.5h feed intervals, with a feed temperature of 33°C. The feed solution was IM each of CaCh and urea with 2.5g of com steep liquor nutrient. The feeding regimen was: 3.6s on / 41.04s off, 25 cycles; System Flow Rate = 100 mL / min; Ave Flow Rate = 2.02 mL / min / brick; and Pulse Volume = 1.50 mL / pulse; to achieve a total volume per feed of 45 mL per cube for a total feed volume for the entire feed process of 720 mL. In these experiments, a pH of 3 gave the best compressive strength, by a comfortable margin, despite the % biocement production scaling with pH. demonstrating that it is not simply a matter of increasing the amount of biocement, but there is an additional factor related to the quality of the biocement produced.

[0046] Figures 7G-7H show the results of an experiment in which the molarity of the feed reagents was varied. The ratio of CaCh to urea was 1:1. The form factor was 125 cc cubes. The fine component of the aggregate was pond fines. The feeding regimen was similar to that of Figures 7E-7F.

[0047] Figure 71 shows the results of an experiment in which the number of feeds was varied with different lengths of holding period (2h to 4h).

[0048] Figure 7J shows an experiment using flexural unit rectangular form factors in which the amount of fine limestone in the aggregate mix was varied. All three dosages produced flexural units with above spec, flexural strengths.

[0049] Figure 7K. shows the results of an experiment in which various pre-feed conditions for biostructures having microorganisms mixed into the aggregate prior to pressing (i.e., ‘mix-in’) were compared to a baseline in which the microorganisms were added topically to the pressed biostructure. The conditions varied were: PIC1 = 1 g / kg yeast extract as nutrient and an incubation duration of 3 h; PIC2a = 1 g / kg yeast extract as nutrient and an incubation duration of 7 h; PIC2b = 2 g / kg yeast extract as nutrient and an incubation duration of 3 h; and PIC3 = 3 g / kg yeast extract as nutrient and an incubation duration of 5 h.

[0050] Figure 7L depicts a system for large scale, high-throughput manufacturing of biostructures using methods described herein (e.g.. the single pass feeding methods).

[0051] Figure 7M shows compressive strengths for a series of experiments having different pre-feed urease activity and CFU. These experiments used the ‘mix-in’ inoculation.

[0052] Figures 8A-8C show the set-up and SEM results of a flask experiment to assess the benefit of acidic feed pH on calcite formation. Figure 8A shows the flask experiment set-up. Figure 8B shows an SEM image of cry stals of calcite formed in the flask experiment with an initial pH of 9. Figure 8C shows an SEM image showing cry stals of calcite formed in flask experiment with an initial pH of 3. In the flask study, a 200 mL solution of calcium chloride and urea was inoculated with urease-producing bacteria and the progress of the reaction was monitored by: change of pH, change of electrical conductivity, and formation of calcite (qualitatively). A high initial pH (pH 9) of the reaction solution was compared to a low initial pH (pH 3) of the reaction solution. Other conditions were the same. The progress of the reactions was monitored by electrical conductivity of the feed solution and by final pH. After 48 h, the resulting calcite formed was characterized qualitatively by scratch resistance with a spatula and by scanning electron microscopy. Despite a smaller increase in electrical conductivity in the low pH experiment, which would be indicative of less biocementation, the resulting calcite formed in the bottom of the was qualitatively very strong (resistive to breaking by spatula), and SEM analysis showed a marked difference in the form of the calcite crystals formed at the microscopic scale. As can be seen by comparing 8B to 8C, the calcite crystals formed in the high initial pH experiment where considerably smaller and more irregular than those formed in the low initial pH experiment. Combined with the qualitative observation of higher resistance to scratching / breaking in the low initial pH flasks, these results suggest that an initial pH that is acidic is beneficial to the production of stronger biocement.

[0053] Figures 9A-9C show the set-up and results of an experiment in which 125 cc cubes were soaked in a feed solution with 1.0 M each of calcium chloride and urea in baths as shown in Figure 9A. The volumetric ratio of feed to brick is controlled by varying the number of bricks soaked in each container (see Figure 9B). Additionally, cubes of the same size and composition were fed in a drip-based feeding system (“microtester”) for comparison. Figure 9C shows the compressive strength results.

[0054] Figures 9D-9H show results from an experiment comparing drip-feeding and soak-feeding in 22 x 50 x 50 mm tiles and 22 x 50 x 200 mm form factors.

[0055] Figures 9I-9K show the set-up conditions and results of an experiment to compare the soak feeding method with the drip feeding method (‘microtester’) using the 125 cc cube form factors. Additionally, a partially fed ‘microtester’ brick was soaked by ending the drip feeding at 4 feed and 8 feeds respectively.

[0056] Figures 9L-9Q show the set-up conditions and the result of experiments investigating various combinations of feed volume : brick volume ratio (F : P ratio), at various molarities (estimated by electrical conductivity (EC)), and soak times. The form factors were 22x50x50 mm units (see table of Figure 9N. Exp IDs A-C) and 22x50x200 mm units.

[0057] Figure 9R shows two full size tiles (22 x 200 x 400 mm) 902 and 903 prepared according to the soak methods described herein. The tiles were made from an aggregate blend that included a 55%-Skygge:45%-Thybo base-blend with a mixture of 2% limestone and 1% Kaolin as added fines. The tiles were cut into multiple smaller units for flexural and compressive strength testing. Tile 902 was subject to a 68-hour soak in a feed solution having 1.3 M each of calcium chloride and urea. Tile 902 had an average flexural strength of 8 MPa and an average compressive strength of 4572 psi. Tile 903 was subject to a 46 hour soak in a feed solution having 1.3 M each of calcium chloride and urea. Tile 903 had an average flexural strength of 4.9 MPa and an average compressive strength of 3313 psi.

[0058] Figure 9S shows a prototype system for producing biostructures, such as tiles 912-913, pavers, blocks, or other 3D structures, using the soak / submersion methods described herein. The recirculation set-up can ensure that all structures (e g. tiles, blocks, pavers, etc.) in separate baths receive the same feed composition, and provides the additional benefit of added mixing and agitation.

[0059] Figure 9T shows a schematic of another prototype system for producing biostructures, such as tiles 912-913, pavers, blocks, or other 3D structures, using the soak / submersion methods described herein. The system may include pH, concentration (EC), and temperature sensors. The system may include one or more fillers, e.g., to reduce the amount of feed solution required and / or to modulate flow of cementation reagent solution. The system includes pumps (e.g., a feed pump, a recirculation pump , and an effluent pump), a feed tank (which may also be a staging stank), an effluent tank, a feedbath and a recirculation valve. The recirculation pump is in fluid communication (controlled by the recirculation valve) with the feed bath and a container for the sensors and recirculates feed between the feed bath and sensors to allow constant monitoring of feed solution properties. The recirculation pump may also generate flow in the feed solution to allow fresh feed solution to reach the biostructures, reducing the degree to which diffusion of reagents controls rate of cementation reagent consumption. The unit container may allow for climate control of the air around the feed bath / feed solution and / or keep the bath clear of contaminants.

[0060] Figure 10A depicts representations of different types of aggregate blend gradation types.

[0061] Figure 10B depicts a representation of fluid flowing through an aggregate blend.

[0062] Figure 10C depicts the results of a study of binary aggregate blends with different ratios of Skygge (‘S’ a type of sand) and Thybo (‘T’, a type of gravel) (i.e., of different gradations). The x-axis is %T (therefore the %S is 100-x) and the two y-axis are (left) density (envelope, g / cc) of the blend and (right) water infiltration rate, Ksat (cm / s). The %water retention (%wt) is also shown, but not associated with either axis, the highest value was 12.86% and the lowest value was 8.56%. The envelope density of the blends peaks at from about S60T40 to about S40T60.

[0063] Figure 10D depicts results of a comparative study of different loadings of basalt fiber additives to the plurality of aggregate particles.

[0064] Figure 10E depicts results of another comparative study of different loadings of basalt fiber additives to the plurality of aggregate particles, including the combination of 0.25 wt% basalt fiber with 2% limestone (in place of pond fines).

[0065] Figure 10F shows photographs of a shaped structure including a strainenhancing textile and steps in its production.

[0066] Figure 10G depicts a schematic of a shaped structure including a strainenhancing textile.

[0067] Figure 10H depicts a graph showing the results of an experiment in which sodium hydroxide was used to fuse limestone in the aggregate blend in a shaped structure before topically inoculating with mineralizing microorganisms and biocementing with MICP.

[0068] Figure 101 depicts a graph showing the results of an experiment in which shaped structures of aggregates and mineralizing microorganisms were incubated at different temperatures after pressing into the shaped structure and before feeding.

[0069] Figure 10J depicts a graph showing the results of an experiment in which shaped structures of aggregates and mineralizing microorganisms were soaked in a bath of feed solution at different temperatures.

[0070] Figure 10K depicts a graph showing the results (flexural strength, in MPa) of an experiment in which shaped structures made from a plurality of aggregate particles inoculated with microorganisms grown in a fermentation solution with clarified com steep liquor as the nutrient were compared to shaped structures made from a plurality of aggregate particles inoculated with microorganisms grown in a fermentation solution with clarified com steep liquor and glucose as the nutrient.

[0071] Figure 11A depicts two types of pallet for holding shaped structures during feeding in a feeding system; on the left a pallet with no holes, on the right, a pallet with holes.

[0072] Figures 11B and 11C depict a climate-controlled curing system (specifically a resting area in the curing system) during manufacturing. Figure B also depicts multiple shaped structures (tiles depicted here) positioned within the curing system / resting area.

[0073] Figures HD to 11H depict a feeding system including multiple spray nozzles attached to a boom that is part of a conveyor system.

[0074] Figure 1 II depicts multiple resting areas of the curing system (foreground) of Figures 1 IB and 11C and the feeding system shown in Figures 1 ID-11H (background). The conveyor system also includes a crane configured to transfer shaped structures between the feeding system and the curing system.

[0075] Figure 11 J the feeding system shown in Figures 1 ID-11H after applying a feed solution to the shaped structures.

[0076] Figure 11 J the feeding system shown in Figures 1 ID-11H after applying a feed solution to the shaped structures.

[0077] Figure UK shows a curing system with a lid and ducts for the climate control system. The incubation chamber is used here as a curing chamber between feeding cycles.but may also be used as an incubation system to allow microorganism expansion and / or crust formation in shaped structures before feeding.

[0078] Figure 11L depicts a schematic of a curing system including a set of climate- controlled resting areas and associated systems.

[0079] Figures 12A-12E show the results of experiments into the effect of gelling agents on surface quality.

[0080] Figures 13 A and 13B show the results of introducing a deflective element between a spray feeding source and the biostructures.

[0081] Figures 14A-14C show the results of introducing a deflective element between a spray feeding source and the biostructures.

[0082] Figure 15 shows a polished tile produced in a spray / gravity-fed system such as described herein.

[0083] Figures 16A and 16B show results of an experiment to compare feeding biostructures held on a pallet with holes vs without holes.

[0084] Figures 17A-17D show results of an experiment to compare feeding biostructures at 1.5 M vs 1 M and with unadjusted pH vs adjusting to pH 3.

[0085] Figures 18A and 18B show results of an experiment to compare different fines.

[0086] Figure 19 shows flexural strength of biostructures made from a Carboniferous Limestone rock-based aggregate blend vs a sandstone rock-based aggregate blend. Also shown is the result of adding 0.5 wt% microspheres and 0.5 wt% PVA fibers to the carboniferous rock aggregate blend.

[0087] Figure 20 shows results of a column experiment showing biocement production in a 1 M, 2 mL per feed feeding protocol vs a 2 M, 1 mL per feed feeding protocol.

[0088] Figure 21 shows pre- and post- Freeze-Thaw test flexural strength of shaped structures fed in a laminar flow / gravity-based feeding system.

[0089] Figure 22 shows the results of a series of experiments to test different additives.

[0090] Figures 23A-23C depict flowcharts describing various embodiments of processes for manufacturing a construction material.

[0091] Figures 24A-24B depict flowcharts describing various embodiments of processes for manufacturing a construction material.

[0092] Figures 25A-25H depict flowcharts describing various embodiments of processes for manufacturing a construction material.

[0093] Figures 26A-26F depict flowcharts describing various embodiments of processes for manufacturing a construction material.

[0094] Figures 27A-27F depict flowcharts describing various embodiments of processes for manufacturing a construction material.

[0095] Figures 28A-28B depict flowcharts describing various embodiments of processes for manufacturing a construction material.

[0096] Figures 29A-29F depict flowcharts describing various embodiments of processes for manufacturing a construction material.

[0097] Figure 30A depicts one embodiment of a biomanufacturing system.

[0098] Figure 30B depicts one embodiment of a biologies activation and / or expansion system.

[0099] Figure 30C depicts one embodiment of a biologies activation and / or expansion system.

[0100] Figure 30D depicts another embodiment of the biomanufacturing system.

[0101] Figures 31 A-31I depict various embodiments of a biomanufacturing system for performing a soak-based feeding process.

[0102] Figures 31J-31P depict flowcharts describing various embodiments of processes for manufacturing a construction material.

[0103] Figures 32A-32I depict various embodiments of a regeneration system for regenerating a feed solution used in a process such as the processes described herein.

[0104] Figure 32J depicts a flowchart describing another embodiment of a process for manufacturing a construction material including a regeneration step.

[0105] Figure 33 depicts a flowchart describing an embodiment of a process for enhancing a construction material.

[0106] Figure 34 depicts a decision flowchart for determining whether to proceed to an inoculation step or continue fermentation (e.g.. expansion) in a liquid medium.

[0107] Figure 35 depicts a pore in a plurality of aggregate particles (e.g., in a shaped structure) and several of the chemical, biological, and material processes that occur therein during urease-based MICP.DETAILED DESCRIPTION

[0108] Technology described herein improves the mechanical and structural properties of construction materials that utilize biocement and reduces the manufacturing time and cost for manufacturing biostructures that include biocement compositions.

[0109] Provided in various embodiments herein are methods of producing construction materials. The methods may include providing (e.g., by flowing or spraying, or by submersion) a cementation reagent solution including one or more cementation reagents to a plurality of aggregate particles (e.g., in a shaped structure, e.g., bricks, pavers, paving tiles, flooring tiles, wall tiles, roofing tiles, decorative tiles, veneers, panels, walls, large pre-cast structures, breeze blocks, cinder blocks, concrete masonry units (CMUs), precast beams, precast columns, wall panels, modular concrete barriers, bollards, architectural cladding, slabs, hollow core slabs, step treads, roof pavers, stone wall caps, decorative stone pavers, stone wall copings, pavestones, stepping stones, solid blocks, hollow blocks, block pads, and retaining wall blocks, concrete blocks (e.g., CMUs), etc.) and forming a mineral binder from the cementation reagents at least in part from activity of an organism and / or an enzyme (e.g., a mineralizing enzyme or organism, e.g., urease or a urease-producing microorganism, or carbonic anhydrase or a carbonic anhydrase-producing microorganism) included in the plurality’ of aggregate particles for a sufficient time to consolidate and / or harden the plurality of aggregate particles, thereby forming the construction material. The cementation reagent solution may have a concentration above 700 rnM, or an acidic pH, or both.

[0110] Provided in various embodiments herein are methods of producing construction materials. The methods may include spraying a volume of a cementation reagent solution including one or more cementation reagents to one or more shaped structures, each including a plurality of aggregate particles, and forming a mineral binder from the cementation reagents at least in part from activity of an organism and / or an enzyme (e.g., a mineralizing enzyme or organism, e.g., urease or a urease-producing microorganism, or carbonicanhydrase or a carbonic anhydrase-producing microorganism) included in the plurality of aggregate particles for a sufficient time to bind two or more particles of the plurality’ of aggregate particles.

[0111] Provided in various embodiments herein are methods of producing construction materials. The methods may include spraying multiple volumes of cementation reagent solution to one or more shaped structures, each including a plurality' of aggregate particles, and forming a mineral binder from the cementation reagents, at least in part from activity' of an organism and / or an enzyme (e.g., a mineralizing enzyme or organism, e.g., urease or a urease-producing microorganism, or carbonic anhydrase or a carbonic anhydrase-producing microorganism) included in the plurality of aggregate particles, for a sufficient time to bind two or more particles of the plurality of aggregate particles. The multiple volumes may be multiple pore volumes. The multiple volumes may vary in volume. The multiple volumes may vary in concentration of cementation reagent and / or pH. The multiple volumes may vary in both volume and concentration of cementation reagent and / or pH.

[0112] Provided in various embodiments herein are methods of producing construction materials. The methods may include submersing one or more shaped structures including a plurality of aggregate particles, into a cementation reagent solution, including one or more cementation reagents, each and forming a mineral binder from the cementation reagents at least in part from activity of an organism and / or an enzyme (e.g., a mineralizing enzyme or organism, e.g., urease or a urease-producing microorganism, or carbonic anhydrase or a carbonic anhydrase-producing microorganism) included in the plurality' of aggregate particles for a sufficient time to bind two or more particles of the plurality of aggregate particles.

[0113] Provided in various embodiments herein are methods of producing construction materials. The methods may include forming a shaped structure including an organism (e.g., a mineralizing organism, e.g., a urease-producing microorganism or a carbonic anhydraseproducing microorganism) and allowing or inducing the organism to proliferate in the shaped structure for a sufficient time to achieve an activity per kilogram of a mineralizing enzyme sufficient to consolidate and / or harden the plurality of aggregate particles when provided (e.g., by methods described herein) with a quantity of cementation reagent sufficient (e.g., such as in one or more cementation reagent solutions described herein) to produce at least 5 wt% of a mineral binder (e.g., a metal carbonate) in the shaped structure,thereby forming the construction material (which may be any construction material described herein).

[0114] Provided in various embodiments herein are methods of producing bioconcrete construction materials by preparing an aggregate blend including two or more aggregates, combining the aggregate blend with mineralizing enzymes or microorganisms, one or more cementation reagents, and forming a biomineral cement to bind the two or more aggregates together. The aggregate blend may be an optimized blend, as described herein, which allows for maximum strength in the finished construction material. For example, an aggregate blend with that produces a maximum density that still permits while permitting sufficient transfer of cementation reagents into a pore network of the aggregate blend. For example, a binary blend. For example, a ternary blend. For example, a gap-graded blend. An aggregate blend may have an unpressed green pycnometric porosity of less than 40% (e.g., of less than 35%, 33%, 34%, 32%, 31%, 30%, 20%, 15%, 10%, etc.).

[0115] Provided in various embodiments herein are methods of enhancing one or more properties of a bioconcrete construction material by applying a coating and / or infiltrating a pore network with materials that block or retard ingress of water or interfere with ice crystal formation.

[0116] Provided in various embodiments herein are methods of producing bioconcrete construction materials with one or more enhanced properties by mixing into their aggregate, prior to biocementation, one or more additives that interfere with ice crystal formation or create pressure-relieving pore structures.

[0117] Provided in various embodiments herein are methods of producing bioconcrete construction materials with one or more enhanced properties by mixing into their aggregate, prior to biocementation. Examples include one or more additives that improve feed infiltration and / or act as gelling agents (e.g., high surface energy gelling agents) to hold a meta-stable shape, solid nutrients that allow for slow release over a feeding process, materials that react, high porosity aggregate particles that provide a network of micropores to relieve tension, etc. Provided in various embodiments herein are bioconcrete construction materials with enhanced properties resulting from the application off a coating and / or infiltration of their pore network with materials that block or retard ingress of water or interfere with ice crystal formation.

[0118] Provided in various embodiments herein are bioconcrete construction materials with enhanced properties resulting from the mixing of their constituent aggregates with one or more additives that interfere with ice crystal formation and / or create pressure-relieving internal pore structures.

[0119] In some embodiments, a process for manufacturing biostructures (e.g., tiles, blocks, or pavers) includes acquiring aggregate particles (e.g., an aggregate mix including both sand and gravel), applying an inoculation composition (e.g., a solid dispersion or liquid suspension of enzymes and / or cells) to the aggregate particles, forming the aggregate particles into a shape (e.g., into a tile, paver, block, or brick shape), determining a feed solution to be applied to the aggregate particles, applying the feed solution to the aggregate particles, and performing a curing process to the aggregate particles subsequent to application of the feed solution. The application of the inoculation solution may be applied before forming the aggregate particles into the shape or after, or both before and after the forming step. In one example, the inoculation solution may only be applied to the aggregate particles prior to forming the aggregate particles into a shape. In another example, the inoculation solution may be applied to the aggregate particles subsequent to forming the aggregate particles into a shape.

[0120] A feed cycle may correspond to application of a particular feed volume (e.g., a volume of liquid comprising a feed solution) to aggregate particles within a particular period of time (e.g., over two hours). A feed solution of the particular feed volume may be applied to a collection of the aggregate particles that have been formed into a shape or structure using a variety of feed delivery' mechanisms, such as soaking the aggregate particles in a bath of a feed solution, spraying a feed solution onto a surface of the aggregate particles, or dripping a feed solution onto a surface of the aggregate particles. In some cases, the particular feed volume (e.g., from 0.5x to 5x, a pore volume of the formed aggregate (e.g., about 0.8-1.2x the pore volume), or a multiple (e.g., lx ,1.5x, 2, 3x. etc.) of the volume of the total volume of the pressed aggregate) may be applied to the aggregate particles using one or more feed pulses. The application of the feed solution may include one or more single-pass feed cycles in which the feed solution applied is not reapplied during subsequent feed cycles. In other cases, the particular feed volume (e.g., l-5x the volume of the formed aggregate) may be applied to the aggregate particles by soaking (e g., fully submersing) the aggregate particles in a bath containing a feed solution. The particular feed volume may correspond to a minimum feed volume (e.g., approximately equal to the pore volume) forthe collection of the aggregate particles such that the amount of the feed solution applied to the aggregate particles is conserved.

[0121] A technical issue with partially or fully submerging a construction material containing aggregate particles in a bath prior to the aggregate particles having been formed into a metastable shaped structure is that the structure may partially disintegrate or experience significant erosion during the feeding process due to lack of a permanent binder (e.g., a mineral binder, or permanent organic binder) binding the aggregate particles together. The issue of the shaped structure partially disintegrating may also occur when a spray-based feeding is applied to a construction material containing aggregate particles if the shaped structure is not in a metastable state. A technical benefit of providing a construction material containing aggregate particles with a metastable shaped structure prior to submersing the aggregate particles in a bath containing a feed solution or prior to applying a spray-based feeding process is that the shaped structure may be maintained during the feeding process. Moreover, construction materials that include cavities (e g., a breeze block or a construction block with cylindrical cavities that traverse a cross section of the construction block) may be manufactured with the cavities preserved.

[0122] A plurality of aggregate particles may be formed (e.g., via a press) into a shaped structure prior to application of a feed solution. A pore volume for the aggregate particles of the shaped structure may comprise a volume that is inside the shaped structure that is available to be filled with fluid. The pore volume may correspond to the void space between the aggregate particles of the shaped structure that is able to be filled with a liquid. A subpore volume for the aggregate particles of the shaped structure may comprise a volume that is less than the total pore volume that is able to be filled with a liquid for the aggregate particles of the shaped structure. In one example, the sub-pore volume for the aggregate particles of a shaped structure may comprise 40-95% (e.g., about 40%, 50%, 60%, 70%, 75%, 80%, 90%, or 95%) of the volume that is inside the shaped structure that is available to be filled with fluid.

[0123] Magnesium sulfate, or a magnesium sulfate rich mineral, may be mixed in to the aggregate particles to allow for the use of aggregate that may otherwise produce an overly caustic environment (e.g., recycled OPC concrete, which can leach calcium hydroxide, or blast furnace slag). Magnesium sulfate will react with hydroxide ions to produce magnesium hydroxide, which has a lower pH than, e.g., calcium hydroxide, helping to maintain an optimal pH for the mineralizing enzymes and / or microorganisms in a biologicalprocess for producing construction materials. An added benefit is that the magnesium hydroxide can also react with carbonate ions, whether produced by the enzymes and / or microorganisms or added by pumping carbon dioxide into the aggregate / feed system, thereby creating additional binder that can be incorporated into the binder matrix produced by the biologies.

[0124] A feed cycle may correspond to application of a predefined amount of biocementation reagents to aggregate particles within a particular period of time. In some cases, the predefined amount of biocementation reagents may be part of a feed solution that is applied to the aggregate particles using one or more feed pulses. In other cases, the predefined amount of biocementation reagents may be part of a feed solution into which the aggregate particles are bathed.

[0125] During the one or more single-use feed cycles, manufacturing parameters such as feed rates, feed pressures, feed rate profiles (e.g.. pulse profiles), feed volume, feed solution pH levels, ambient temperature, and ambient humidity may be set and / or adjusted for each of the one or more single-use feed cycles to reduce manufacturing time and to improve the structural integrity of a resulting construction material. Prior to performing the one or more single-use feed cycles, the type and percentage of a binder within an aggregate mix of the aggregate particles may be determined based on ambient temperature and humidity. Furthermore, prior to performing the one or more single-use feed cycles the shaped structure may be held (or incubated) for a period of time between pressing and feeding to allow for further grow th of microorganisms (which may also include partial hardening to metastability, e.g., by activation of a pre-feed binder), e.g., in a pre-feed incubation period, e.g., with pre-feed incubation parameters (e.g., ambient temperature and humidity and / or activation of a pre-feed binder (e.g., a pre-feed mineral binder, e.g., using other binding chemistries described herein (e.g., adding carbon dioxide to react with brucite), or an organic (e.g., polymer) binder, e.g., cornstarch or alginate), e.g., using heat, humidity, dry air, light (e.g., UV), a chemical initiator, etc.)). Furthermore, prior to performing the one or more single-use feed cycles a pre-feed crust layer may be formed (e.g., by activation of the binder by surface drying, and / or e.g., activation of a pre-feed mineral binder, e.g., using other binding chemistries described herein (e.g., adding carbon dioxide to react with brucite), or an organic (e.g., polymer) binder, e.g., using heat, humidity, dry air, light (e.g., UV), a chemical initiator, etc.) to increase the structural integrity of a formed shape of the aggregate particles during an initial set of feed cycles. The thickness ofthe pre-feed crust layer may be determined based on ambient temperature and humidity during a pre-feed holding period (e.g., a pre-feed incubation period). A particular thickness of the pre-feed crust layer may be achieved by modulating the ambient temperature or humidity, or applying airflow, during a pre-feed holding period (e.g., a pre-feed incubation period). The conditions (temperature, time, humidity, airflow, etc.) of the pre-feed holding period may be optimized to achieve both a desired crust formation and a desired number of doublings of the microorganisms. The conditions of the pre-feed holding period may be selected or controlled to maintain an internal moisture content that causes the microorganisms to remain in a vegetative cell state.

[0126] A feed rate profile may specify feed rates of a feed solution applied to a construction material (e.g., comprising aggregate particles) during a feed cycle. In some cases, a feed rate profile or the rate that the feed solution is applied to a surface of the construction material may be dynamically adjusted over time or set every feed cycle. The rate that the feed solution is applied to a surface of the construction material may be set / reset in response to a measured output (e.g., the residual pH, concentrations of residual feed reagents or cementation reaction products / byproducts (e.g., CO2, NH3, NH4CI, etc.)). In one example, during a feed cycle, a fluid delivery system (e.g., a spray system) may apply the feed solution (e.g., cementation solution) to the surface of the construction material at a first feed rate (e.g., at 10 ml per minute) during a first time period and then apply the feed solution to the surface of the construction material at a second feed rate (e.g.. at 5 ml per minute) during a second time period subsequent to the first time period. The feed solution may be applied to the surface of the construction material using a single-use feed cycle that does not use a recirculated solution. The first feed rate and the second feed rate may be set based on ambient temperature, ambient humidify, pH of the feed solution, retained volume, overspill volume, and / or pH or feed component concentration or reaction product concentration of the residual feed solution that includes a portion of the feed solution that is collected after application of the feed solution to the construction material. Alternatively, or in addition, the first and second (and subsequent) feed rates may be set based on a predetermined adjustment based on the feed number, e.g., reducing the feed volume based on the number of preceding feeds. Alternatively, or in addition, the feed volume may be adjusted based on a detected infiltration rate, e.g., by detecting a change in the reflectivity of the surface of a shaped structure (e.g., with a light source configured to shine on one or more shaped structures in the feed system, and optical sensor configured to detect a changein light reflect from the one or more shaped structures), and adjusting based on a time, or a change in the time, from feed application to the change in reflectivity. A technical benefit of adjusting the feed rate during manufacturing is that clogging related issues may be reduced, the chemical efficiency of the feeding process can be increased, the overall cost to manufacture the construction material may be reduced, the compressive strength of the construction material may be increased, the flexural strength of the construction material may be increased, the process efficiency and throughput may be increased, and the manufacturing variability in construction material properties may be reduced.

[0127] In some embodiments, the temperature of the construction material, the temperature of the feed solution, the molarity of one or more cementation reagents in the feed solution, the pH of the feed solution, and / or the ambient humidity7of the environment in which the construction material is arranged during one or more feed cycles may be varied during manufactunng of the construction material. In one example, the temperature of the feed solution may be less than the temperature of the construction material. In one example, the temperature of the feed solution may be greater than the temperature of the construction material. In one example, the pH of the feed solution may include a low pH feed solution (e.g., a pH of less than 7, e.g. less than 6, e.g., less than 5. e.g., less than 4, e.g., about 2-4, e.g., about 2, 3, or 4). The pH level of the feed solution corresponds to how acidic or alkaline the feed solution is. During the formation of calcite within the construction material, the temperature of the construction material and the humidity of the environment in which the construction material is situated may be adjusted to improve the resulting compressive strength of the construction material. In some cases, during a first feed cycle, a feed solution of a first molarity and a first pH may be applied to construction material, then during a second feed cycle subsequent to the first feed cycle, a feed solution of a second molarity7less than the first mortality and a second pH greater than the first pH may be applied to the construction material. During subsequent feed cycles, the molarity of the feed solution applied to the construction material may change from a high molarity solution to a low molarity7solution while the pH of the feed solution may change from a low pH solution (e.g., about pH 3) to a less acidic pH solution (e.g., about pH 4-6, e.g., about pH 4, 5, or 6), to an approximately neutral (i.e., about pH 7), or to a high pH solution (e.g., pH 8-9). Alternatively, the pH may be reduced overtime, e.g., to about pH 3, e.g., to reduce clogging or to encourage the dissolution and reprecipitation of precipitated calcite, leading to a greater fraction of larger cry stals. In some cases, during a first feed cycle, a feed solution of a firstmolarity and a first pH may be applied to construction material, then during a second feed cycle subsequent to the first feed cycle, a feed solution of a second molarity greater than the first mortality and a second pH lower than the first pH may be applied to the construction material. During subsequent feed cycles, the molarity of the feed solution applied to the construction material may change from a low molarity solution to a high molarity solution while the pH of the feed solution may change from a low pH solution (e.g., about pH 6) to a more acidic pH solution (e.g.. about pH 2-3, e.g.. about 3).

[0128] In some embodiments, prior to performing a set of feed cycles (e.g., prior to performing a set of ten feed cycles), a crust formation process may be performed to improve the structural integrity of the green construction material. A technical benefit of performing the crust formation process prior to applying a feed solution to the green construction material is that a formwork may not be required for the construction material during manufacturing, thereby reducing manufacturing costs. A technical benefit of performing the crust formation process prior to applying a feed solution to the green construction material is that the crust formation can reduce shedding, drill-out, wash-out, and pitting. A further technical benefit is to allow for submersion-based feeding. A further technical benefit is to reduce losses of green structures while handling (e.g., during conveyance) prior to feeding. A feed cycle may include a first time period (e.g., three to 10 minutes) during which a feed solution is applied to a construction material (e.g., continuously, or according to a pulse profile as described herein) and a second time period (e.g., two to four hours, e.g., three hours) during which a feed solution is not applied to the construction material. Biocementation can occur during both the first and second time periods. In some embodiments, the construction material is allowed to dry out during the second time period. In other embodiments, the local environment is controlled to inhibit moisture loss during the second time period. In some embodiments, water is added (e.g., by spray ing) during the second time period.

[0129] In some embodiments, fine grain aggregate particles (e.g., limestone particles, clay, crushed glass, fine sand, or combinations thereof) may be used in place of pond fines within the construction material. An aggregate mix may include a mixture of sand, gravel, crushed stone, crushed glass, clay, and / or fine grain limestone particles that have diameters less than 250 micron or less than 100 micron. The fine grain limestone may have a mean particle diameter (or greatest cross-sectional dimension, or smallest cross-sectional dimension) of less than 250 micron, e g., between 1 micron and 250 micron (e.g., between1 micron and 2 micron, between 1 micron and 5 micron, between 2 micron and 4 micron, between 2 micron and 3 micron, between 1 micron and 10 micron, between 2 micron and 10 micron, between 5 micron and 10 micron, between 1 micron and 20 micron, between 10 micron and 20 micron, between 1 micron and 100 micron, between 10 micron and 100 micron, between 1 micron and 50 micron, between 15 micron and 25 micron, between 20 micron and 40 micron, between 20 micron and 50 micron, between 25 micron and 50 micron, between 50 micron and 100 micron, between 30 micron and 100 micron, between 40 micron and 80 micron, between 25 micron and 75 micron, between 50 micron and 150 micron, between 75 and 150 micron, between 80 and 120 micron, between 125 micron and 225 micron, between 150 micron and 250 micron, between 175 micron and 225 micron, between 180 micron and 220 micron, between 200 micron and 220 micron, or between 200 and 250 micron) e.g., less than 225 micron (e.g., less than 200 micron, less than 150 micron, less than 100 micron, less than 50 micron, less than 20 micron, less than 10 micron, less than 5 micron, or less than 2 micron) e.g., about 1 micron, about 2 micron, about 5 micron, about 10 micron, about 15 micron, about 20 micron, about 30 micron, about 40 micron, about 50 micron, about 60 micron, about 70 micron, about 80 micron, about 90 micron, about 100 micron, about 120 micron, about 150 micron, about 175 micron, about 200 micron, about 220 micron, or about 250 micron. The fine grain limestone may have a median particle diameter (or greatest cross-sectional dimension, or smallest cross-sectional dimension) of less than 250 micron, e.g.. between 1 micron and 250 micron (e.g.. between 1 micron and 2 micron, between 1 micron and 5 micron, between 2 micron and 4 micron, between 2 micron and 3 micron, between 1 micron and 10 micron, between 2 micron and 10 micron, between 5 micron and 10 micron, between 1 micron and 20 micron, between 10 micron and 20 micron, between 1 micron and 100 micron, between 10 micron and 100 micron, between 1 micron and 50 micron, between 15 micron and 25 micron, between 20 micron and 40 micron, between 20 micron and 50 micron, between 25 micron and 50 micron, between 50 micron and 100 micron, between 30 micron and 100 micron, between 40 micron and 80 micron, between 25 micron and 75 micron, between 50 micron and 150 micron, between 75 and 150 micron, between 80 and 120 micron, between 125 micron and 225 micron, between 150 micron and 250 micron, between 175 micron and 225 micron, between 180 micron and 220 micron, between 200 micron and 220 micron, or between 200 and 250 micron) e.g., less than 225 micron (e g., less than 200 micron, less than 150 micron, less than 100 micron, less than 50 micron, less than 20 micron, less than 10 micron, less than 5 micron, or less than 2 micron) e.g., about 1 micron, about 2 micron, about 5 micron,about 10 micron, about 15 micron, about 20 micron, about 30 micron, about 40 micron, about 50 micron, about 60 micron, about 70 micron, about 80 micron, about 90 micron, about 100 micron, about 120 micron, about 150 micron, about 175 micron, about 200 micron, about 220 micron, or about 250 micron. In some embodiments, the fine grain limestone may have a particle size distribution in which 100%, 90%, 80%, 75%, 70%, 60%, 50%, 40%, 30%, 25%, 15%, or 10% of the particles are smaller than 20 micron. The fine grain limestone may have a particle size distribution in which 100% of the particles pass through a mess of with a mesh size of 2000 micron, 125 micron, 63 micron, or 45 micron. The fine grain limestone may have a particle size distribution in which 99% of the particles pass through a mess of with a mesh size of 250 micron, or 20 micron, or 10 micron. The fine grain limestone may have a particle size distribution in which 97% of the particles pass through a mess of with a mesh size of 125 micron, or 63 micron, or 10 micron. The fine grain limestone may have a particle size distribution in which 50% of the particles pass through a mess of with a mesh size of 2 micron. The fine grain limestone may have a particle size distribution in which 18% of the particles pass through a mesh with a mesh size of 1 micron. The fine grain limestone may have a Mohs hardness of between 2 and 4 (e.g., 2- 2.2, 2-2.4, 2-2.5, 2-2.6, 2-2.8, 2-2.9, 2-3, 2-3.2, 2.2-3.2, 2.5-3.5, 2.5-3, 2.8-3.8, or 3-4, e.g., about 2.2, 2.5, 2.7, 2.8, 3, 3.2, 3.5, or 4). The fine grain limestone may have a porosity of from 0.1% for the to 40% (e.g., from 0.1 to 1%, from 1% to 10%, from 10% to 40%, e.g. from 20% to 40%, from 25% to 40%. from 30% to 40%. from 35% to 40%, or from 38% to 40%. Alternative fine-grained components, e.g., having particle size distributions as described herein, may include Wollastonite, Bentonite, gypsum (e.g., plaster of Paris), pond fines, pozzolans (e.g., fly ash or volcanic pozzolans), etc. Some fine-grained aggregate components may also act as binders (e.g., in place of, or complementary to, a gelling agent such as cornstarch, tapioca starch, potato starch, wheat starch, inulin, microcrystalline cellulose, guar gum, sodium alginate, pectin, gelatin, polyvinyl alcohol, xanthan gum, etc.). In some embodiments, the gelling agent is a high surface energy' gelling agent, e.g., alginate.

[0130] In some cases, solid state bioprocesses (e.g., germination, activation, expansion, proliferation, femientation, etc.) may reduce the amount of inoculation solution applied to a construction material prior to performing a feed cycle.

[0131] In some cases, a feed solution does not include nutrients for the mineralizing (e.g., urease producing or carbonic anhydrase-producing) microorganisms. A technical benefit of not including the nutrients for the mineralizing (e.g., urease producing or carbonicanhydrase producing microorganisms) is that the amount of contamination in a feed tank storing the feed solution may be reduced. For example, as the feed tank may not be a sterile environment, not including the nutrients for mineralizing (e.g., urease producing or carbonic anhydrase producing) microorganisms within the feed solution may starve out competing microorganisms or prevent other microorganisms from taking resources from the mineralizing (e g., urease producing or carbonic anhydrase producing) microorganisms within the feed tank.

[0132] In some embodiments, a feed system (e.g., a feed delivery system) for applying a feed solution to the surface of a construction material may be controlled to apply the feed solution between a maximum feed rate and a minimum feed rate. The feed system may include a spray system or a drip-based system. A minimum feed rate may be required to ensure that the entire surface of the construction material is coated. In this case, if less than the minimum feed rate is applied, then only a fraction of the surface of the construction material may be coated. Applying a minimum feed rate may also have the advantage of causing a uniform pool of liquid (e.g., cementation reagent solution / feed media) to form (e.g., allowing enough feed solution to a surface of the shaped structure to form a convex meniscus), which has been observed to improve uniformity of flow through the green construction material (for example, where surface tension from the meniscus acts as an additional driving force to push feed solution into the pores). A maximum feed rate may be required to ensure that structural damage (e.g., wash-off, drill-out, pitting, etc.) does not occur to the surface of the construction material due to excessive feed pressure or material erosion. Moreover, the minimum feed rate may enable a spray pattern for a fluid delivery system to match the surface profile of the construction material. A feed rate or feed pressure for the fluid delivery system that is less than a threshold may cause portions of the surface of the construction material to be unable to receive the feed solution. Moreover, a fluid delivery system with nozzles may require at least a minimum nozzle pressure to guarantee an accurate feed rate through the nozzles.

[0133] In some cases, a sealant may be applied to the surface of a construction material during a finishing process to prevent acidic erosion and to prevent water from penetrating the surface of the construction material. Moreover, an ozone (or other oxidative) treatment may be performed to remove odor from ammonia gas produced within the construction material and / or to destroy any remaining viable microbial cells.

[0134] As used herein, the term "construction material” or “construction materials” generally refers to an article which includes elements or subcomponents (e.g., aggregate) that are bound together by linkages (e.g., cement linkages) or bridges of adhesive properties. In some cases, the linkages or bridges within a construction material include a binding agent, such as calcium carbonate. Construction materials may include individual physical objects of a defined shape that are incorporated into, for example, a building, a structure, or a work.

[0135] Some construction materials may include biocement products. Examples of biocement products include, but are not limited to, items made from bioconcrete, biocement coated aggregates, and the like. As used herein, the term “biocement” generally refers to any binding agent that can be generated through a biological mechanism (e.g., an enzymatic process) which adheres to or encapsulates particles of solid material (e.g., aggregate particles). The binding agent may directly connect two or more aggregate particles, or may indirectly connect two or more aggregate particles via one or more linkages or bridges within a construction material. The one or more linkages or bridges within the construction material may provide structural support for the construction material and / or provide a structural connection between at least two particles of a plurality of aggregate particles within the construction material. In some cases, the aggregate particles include sand, crushed stone, and / or gravel. The shape of the aggregate particles may be classified as either angular, subangular, subrounded, or rounded. Examples of a biocement include calcium carbonate bound to a preexisting particle of a solid material which was formed by microbiologically induced carbonate precipitation (MICP). Further examples include biologically sintered metal carbonates, such as but not limited to calcium carbonate, magnesium carbonate, barium carbonate, or strontium carbonate. Another example is dolomite.

[0136] As used herein, the term “bridging calcium carbonate” generally refers to calcium carbonate that is between, connects with, and / or contributes to the binding together of at least two moieties. such as aggregate particles. Together with the at least two moieties, the bridging calcium carbonate provides for a continuous piece comprising the at least two moieties and the bridging calcium carbonate. The calcium carbonate may be a solid, such as a precipitate. The calcium carbonate may be formed from the reaction of calcium ions with carbonate ions in aqueous solution, wherein the resulting calcium carbonate is integrated into a bulk composite material comprising calcium carbonate and the aggregate particles. Calcium carbonate that does not bridge or connect two pieces of aggregate canstill be considered bridging if it contributes to the strength of the overall binding, e.g., by further structurally reinforcing a bridging or connecting piece of calcium carbonate that it is attached to either directly or indirectly.

[0137] As used herein, the term "non-bridging calcium carbonate” generally refers to calcium carbonate that does not connect with or contribute to the binding together of at least two moi eties, such as aggregate particles. The non-bridging calcium carbonate may be a precipitate that connects with only one moiety, such as an aggregate particle, or a precipitate that does not connect with any aggregate particle. The non-bridging calcium carbonate may be bound to at most one moiety.

[0138] As used herein, the terms "aggregate” or "aggregate particles” may be used interchangeably and generally refer to any type of particulate matter which can be bound together into larger particles or consolidated solids by biocement bonds or bridges. Nonlimiting examples of aggregates include sand, crushed stone, mine tailings, or combinations thereof, etc.

[0139] As used herein, the terms “fines” or “fine component” or “fine” as an adjective for particles (e.g.. mineral particles, e.g., limestone) is used to mean particles that are in the micron scale, e.g., having a diameter less than 250 microns. Such particles may be used as performance enhancing additives, as fillers, to form or enhance a binder, as supplementary cementitious materials, etc. A fine fraction of an aggregate or a fine aggregate (e.g., sand) of an aggregate blend (as opposed to a "coarse aggregate”, e.g., gravel) may also include particles of such sizes, but may also include particles of up to 4.57 mm in diameter.

[0140] As used herein, the terms "single-pass” and "single-use” refer to feeding processes and systems where a feed solution is not recirculated back into a plurality’ of aggregate particles without being modified in some way (e.g., regenerated using a regeneration system described herein, e.g.. re-concentrated by addition of fresh reagents), or is not recirculated back at all.

[0141] As used herein, the term “cementation reagents” generally refers to any combination of starting matenals, which when combined and allowed to react produce a binding agent via a biological mechanism. For example, cementation reagents in a ureahydrolysis based biocementation system may include urea (or another suitable nitrogen source), a soluble calcium source (e.g., calcium chloride, calcium acetate, calcium phosphate, calcium sulphate, calcium lactate, calcium nitrate, etc.). Cementation reagentsmay also include an enzyme (or an organism containing an enzyme) can lead to the enzymatic formation of a biocement, such as calcium carbonate, which binds together adjacent aggregate particles. The cementation reagents may also include nutrients which promote urease activity (which may vary depending on whether pure enzyme or ureaseproducing cells are used), and urease to form and precipitate a calcium carbonate biocement. In a calcium carbonate based biological sintering biocementation system, examples of biocementation reagents can include calcium carbonate, nutrients which promote enzymatic acid production, and an acid producing enzyme which generates acid to dissolve the calcium carbonate. Biocementation reagents for such a system may also include a second set of nutrients and a second enzy me which together promote a pH drop, reprecipitating calcium carbonate to form a biocement.

[0142] As used herein, the terms "‘produce,” “production,” and “producing” regarding a mineral binder (e.g.. a metal carbonate, e.g.. calcium carbonate) in the presence of an enzyme or a biological organism refer to the biological reactions enabled by the enzyme or the biological organism to produce the conditions to form the binder (e.g., calcium carbonate) from starting materials such as a metal (e.g., calcium, magnesium, etc.) ion, carbonate ion. or other possible chemical entities. For example, an enzyme, which produces carbonate ion or calcium ion, can be called an enzyme producing calcium carbonate. An enzyme, which causes pH changes to enable the precipitation of a mineral binder (e.g., calcium carbonate), can also be called an enzyme producing the mineral binder (e.g., calcium carbonate).

[0143] As used herein, the terms Microbiologically Induced Carbonate Precipitation (MICP) (also known as Microbially Induced Calcite Precipitation) generally refers to the production of metal (e.g., calcium) carbonate using at least one enzyme or biological organism. The at least one enzyme or biological organism can form either calcium ion or carbonate ion, or can change the pH of the environment to precipitate the metal (e.g., calcium) carbonate. In the methods described herein, MICP may also produce carbonates of other metals, which may or may not be incorporated into the bridging calcium carbonate.

[0144] The construction materials described herein can take many different forms and shapes. An example construction material, such as a unit (e.g., a brick, tile, block, or paver) with a top surface, four vertical sides and a bottom surface, may be used to illustrate the many features of the disclosed embodiments. However, the construction materials are not limited to only cubiform. In one example, a construction material may include a tile, suchas a round tile with a top surface, a vertical circular side, and a bottom. In another example, a unit may be formed (e.g., pressed) in a mold or formwork that gives it one or more curved faces, edges, cavities, hollows, or vertices.

[0145] Structural biocement-based concrete (e.g., a biocement brick) may rely on the construction of interconnected crystals of a mineral binder (e.g., calcium and / or magnesium carbonate crystals, metal phosphate crystals, silicate crystals, etc.) to bind together surrounding aggregate particles. While biocement formation may be the product of Microbiologically Induced Calcite formation, MICP may result in calcium carbonate not related to the formation of structural biocement (e.g., an isolated calcium carbonate crystal that does not bind to more than one aggregate).

[0146] In some cases, connected biocement bridges may form as homogeneously as possible throughout the aggregate substrate / network, binding neighboring aggregate particles, thereby ensuring structural integrity throughout the product (e.g., one area may not be much stronger or much weaker than another). In some cases, the structural integrity of the product may be achieved by distributing MICP within the body of the biocementbased concrete such that the concentrations or weight percentages of the bridging calcium carbonate or calcium to silicon ratios at different cross-sections of the product differ no more than 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%. 30%, 25%, 20%, 15%, or 10% from each other.

[0147] When a MICP reaction forms non-connected. isolated, non-bridging calcium carbonate crystals, these crystals can fill pore-spaces between aggregate particles, preventing additional calcium carbonate formation required for structural bridging, thereby called clogging. Furthermore, when a solution containing calcium and urea is directionally applied to an aggregate substrate, if biocement is produced at a faster rate at the surface of solution application (the non-preferred surface), pore-space reduction by biocement formation or clogging can prevent biocement formation deeper within the unit / biocement- based concrete. In some cases, when homogenous biocementation (e.g., due to low7variations of the degrees of MICP formation within the body of the unit) is difficult to achieve for a unit / biocement-based concrete, another goal can be to produce biocement at higher concentrations at the surface (the preferred surface) furthest away from the surface of solution application. The formation of non-structural, non-connected calcium carbonate crystals concentrated at the surface of the material can occur w hen pore-spaces are reduced to the point that the solution applied can no longer enter the inside of the unit. Thisphenomenon is described as a "crash -out", which denotes formation of calcium carbonate on the surface of the unit. Preventing crash-out is desirable in practice to reduce the cost of direct materials, while improving product performance, consistency, and manufacturing repeatability. The early-stage formation of crash-out is an indicator that biocement or clogging has occurred first at the non-preferred surface (or the surface of solution application).

[0148] In some embodiments, preventing or reducing crash-out in practice is achieved (a) if the solution applied is free of particulate matter that can clog aggregate pore-spaces in a non-structural manner, and / or (b) the rate of cementation is not faster (or is slower) at the non-preferred surface (or the surface of solution application) or the rate of cementation is below a rate threshold.

[0149] Regarding (a): after searching for conditions that may reduce the formation of non-bridging calcium carbonate in a crash-out. the following conditions are identified: use filtration for the feed solution to prevent inert materials such as loose aggregate and / or plant matter from circulating within the supplied calcium solution and / or urea solutions. However, although feed solutions can be practically non-sterile, contamination from microorganisms in the aggregate matrix may contaminate the solution as free-floating cells. When MICP occurs outside the network of aggregates but in the solution surrounding the network of aggregates, discrete, isolated, non-connected calcium carbonate crystals are formed, which may also create particulate matter within the solution outside the aggregates. Therefore, it may also be beneficial to inhibit / reduce biological activity7(generating enzymes or biological organisms) within the solution, so that MICP primarily occurs only within the aggregate substrate in which biocement formation is desired. In practice, biological activity can be successfully inhibited with the application of UV radiation treatment and / or the induction of ozone and / or the blow of fresh air over the top of solution outside the aggregate network.

[0150] Regarding (b): one option is to use environmental and process controls to ensure the non-preferred surface may not build biocement faster than the preferred surface does. In some cases, the non-preferred surface is at the top surface as the feed solution may be applied from a fluid delivery system that relies on gravity' to apply the feed solution. The rate of biocement formation may correlate with both the metabolic activity of the microorganisms or enzymes, and their rates of propagation (e.g., fermentation, doubling rate). Temperature, nutrient-availability, and respiration may influence biological activityof the microorganisms or enzymes. Therefore, options include (a) control the temperature of the process such that the temperature at or near the non-preferred surface is not higher than (or is less than) the temperature at or near the preferred surface, which may be controlled by ensuring that the aggregate substrate is warmer than the solution that is applied to the non-preferred surface; (b) nutrient in the feed solution is fully dissolved so that it can be homogeneously dispersed throughout the unit: and / or (c) in the case of anaerobic or facultatively-anaerobic microorganisms used for the MICP process, ensure that ample oxygen is available for respiration of the microorganisms, which may be achieved through regular air changes in a closed system, or natural ventilation in an open system.

[0151] As used herein, the unit “g / cc” generally refers to the unit grams per cubic centimeter (g / cm?), which is a unit for density. The percentage of void within a portion of a construction material may correspond to an average porosity for the portion of the construction material.

[0152] As used herein, the term “green density’7generally refers to the density of the construction material before sintering. The term “green density" may refer to wet green density, which may include the density of a pressed unit at the time of pressing, or to dry green density, which may include the density of a pressed unit that has been completely- dried out.

[0153] As used herein, the term “finished density’’ generally refers to the density of the construction material after sintering (i.e., after the formation of mineral binder(s). e.g., biocement (e.g., metal carbonates, phosphates, silicates, etc., produced by or with the action of a mineralizing enzyme or microorganism) and any other non-biocement cementitious binders (e.g., pozzolanic binding, fusion of carbonates with metal hydroxides, etc.)). In some embodiments, the average green density is from about 1.5 g / cc to about 1.8 g / cc. from about 1.6 g / cc to about 1.9 g / cc, from about 1.7 g / cc to about 2.0 g / cc, from about 1.8 g / cc to about 2.1 g / cc, from about 1.9 g / cc to about 2.2 g / cc, from about 2.0 g / cc to about 2.3 g / cc, from about 2. 1 g / cc to about 2.4 g / cc. In some embodiments, the average green density is from about 1.7 g / cc to about 2.0 g / cc. In some embodiments, the average green density is about 1.5. 1.6. 1.7, 1.8, 1.9, 2.0. 2.1, 2.2, 2.3, or 2.4 g / cc. In some embodiments, the average finished density is from about 1.8 g / cc to about 1.9 g / cc, from about 1.9 g / cc to about 2.0 g / cc, from about 2.0 g / cc to about 2.1 g / cc, from about 2. 1 g / cc to about 2.2 g / cc, from about 2.2 g / cc to about 2.3 g / cc, from about 2.3 g / cc to about 2.4 g / cc, from about 2.4 g / cc to about 2.5 g / cc, from about 2.5 g / cc to about 2.6 g / cc. or from about 2.6 g / cc to about 2.7 g / cc. Insome embodiments, the average finished density is from about 2.0 g / cc to about 3.1 g / cc (e.g., 2.0 g / cc to about 2. 1 g / cc). In some embodiments, the average finished density is about 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6 or 2.7 g / cc. In some embodiments, the average finished density' is greater than 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, or 3 g / cc. In some embodiments, the average percentage of void of green density is from about 25% to about 28%. from about 28% to about 31%, from about 31% to about 34%, from about 34% to about 37%. from about 37% to about 40%. from about 40% to about 43%, or from about 43% to about 46%. In some embodiments, the average percentage of void of green density (i.e., the average percentage of void in a green structure) is from about 30% to about 42%. In some embodiments, the average percentage of void of green density' is about 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%. 35%. 36%. 37%. 38%, 39%, 40%, 31%, 42%, 43%, 44%, 45%, 46%, or 47%. In some embodiments, the average weight percentage of CaCCh made to turn a green structure into a finished structure is from about 5% to about 30% e.g., about 5% to about 10%, from about 10% to about 15%, from about 15% to about 20%, or from about 20% to about 25%. In some embodiments, the average weight percentage of CaCOs made is from 10% to about 23%. In some embodiments, the average weight percentage of CaCCh made is about 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, or 25%. In some embodiments, the average percentage of CaCCh made to make a finished structure is less than 20%, e.g.. less than 15%, e.g.. less than 10%. In some embodiments, a difference between the finished density and the green density is primarily due to the formation of calcium carbonate precipitated from a soluble calcium source in the feed solution.

[0154] The aggregate material may include rock (e.g., crushed rock, e.g., crushed to gravel, or sand, or fines, or a combination thereof), sand, glass, wood, paper, metal, plastic, polymers, minerals, manufacturing or processing waste materials such as ash, carbon, or wood residuals, any of which can be crushed or used whole or combinations thereof. For some structures, sand and fines may be overlapping concepts, e g., a fine aggregate fraction of a blend may include . For some structures, a course fraction of an aggregate blend includes stones, crushed rock, etc., that are also called gravel.

[0155] The aggregate material may include organic or inorganic material such as, for example, sand, rock, glass (e.g., Poraver), wood, biochar, paper, metal, plastic, polymers, minerals, recycled materials, or combinations thereof. Aggregate particles may include beads, grains, rods, strands, fibers (e g., fiberglass, basalt fibers, jute fibers, polymer (e.g.,polyethylene, polypropylene, cross-linked polyvinyl alcohol, or POM) fibers, etc ), flakes, crystals, pulverized or crushed materials, or combinations thereof. The construction material may include bricks, thin bricks, pavers, panels, tile, veneer, concrete masonry units (CMUs) (e.g., cinder, breeze, besser, clinker, or aerated blocks), counter- or table-tops, design structures, blocks (e.g., hollow blocks), a solid masonry structure, piers, foundations, beams, walls, slabs, or combinations thereof. The construction material may be a solid 3D shape (e.g., a cuboid) or may include cavities (i.e.. hollows), e.g.. a cuboid with one or more cavities (e.g., a brick with one or more (e. g., 1, 2, 3, or 4) cylindrical cavities that traverse a cross section, or hollow depression(s) in one or more faces of the brick, or, e.g., a hollow concrete block).

[0156] Aggregate particles may have a diameter (e.g., actual, average, or effective diameter) of about 50 mm or less, preferably about 25 mm or less, preferably about 20 mm or less, preferably about 10 mm or less, and preferably about 5 mm or less. In some embodiments, aggregate material may be about 1 mm or less and about 0.5 mm or less, about 0.1 mm or less, and about 50 pm or less. Particles sizes may include from about 10 pm to about 1 mm, from about 100 pm to about 0.5 mm, from about 200 pm to about 1 mm, from about 1 pm to about 200 pm, from about 10 nm to about 1 pm. and from about 10 nm to about 40 nm, and various combinations thereof. Aggregate particles may be largely composed of particulates of less than 5 mm in diameter (e.g., less than or about 4 mm, less than or about 3 mm. less than or about 2 mm, or less than or about 1 mm). In some embodiments, an aggregate blend may be larger, for example, limited to having a greatest diameter that is no larger than 0.25x a shortest cross-sectional dimension (e.g., a thickness) of a construction material (e.g., having a maximum particle diameter (e.g., a highest sieve size) that is about 0.25x a thickness or a tile or paver to be made).

[0157] Aggregate particles may be characterized by fine size and may be equal to or less than 250 micron, equal to or less than 200 micron, equal to or less than 150 micron, or equal to or less than 100 micron (reference examples include micron size of beach sand = 700, micron size of fine sand=250; micron size of Portland cement = 74; micron size of silt = 44; micron size of smoke = 2).

[0158] In some embodiments large particles may be used, e.g., aggregates comprising particles of cm or mm size, e.g., gravels, stones, crushed rocks etc. In some embodiments, the size range of particles may range from 1 pm, or 10 pm, to 5 cm, or more. In some embodiments, the particulate starting material may consist of, or at least essentially consistof, small particles, in particular particles on a pm scale. In other words, the particles may be between, or in the range of 1 pm and 1 mm (1000 pm) in size, for instance between, or in the range of, 100 and 1000 pm or 100 and 500 pm, for example 200 and 400 pm, or 200 and 300 pm. Large size particles may however be, for instance, between 1 mm and 2 mm in size, or have a wider range in size, for instance between 100 pm and 2 mm.

[0159] The bridging calcium carbonate may bond to individual aggregate particles to create bridges between the aggregate particles, or it may encapsulate individual aggregate particles. The bridging calcium carbonate may be formed from the reaction of calcium and carbonate ions. The mineralizing enzyme or mineralizing microorganism and the reagents may be added together or separately to result in the formation of calcium carbonate. The calcium carbonate may be a precipitated calcium carbonate. In some embodiments, the bridging calcium carbonate may be crystals, such as bridging calcium carbonate crystals. In some embodiments, the bndging calcium carbonate crystals are ordered, pseudocrystalline, or amorphous. In some embodiments, the bridging calcium carbonate cry stals are macroscopically ordered and include a trigonal, orthorhombic, or hexagonal crystal structure.

[0160] The at least one biological organism or enzyme may include a mineralizing (e.g., urease-producing or carbonic anhydrase producing) microorganism. The urease-producing microorganism may be an organism in the domains of archaea (e.g.. haloarchaea), bacteria (e.g., Cyanobacteriota (e.g., cyanobacteria), Pseudomonadota, Gammaprobacteria, Firmicutes, Campylobacterota, Myxococcota), or eukarya (e.g., fungi (e.g., yeasts (e.g., of the genus saccharomyces) or filamentous fungi (e.g., of the genus Aspergillus)), protists (e.g., diatoms or coccolithophores). and some algae (e.g., Thraustochytrids)). Ureaseproducing bacteria include organisms in the phyla Pseudomonadota (e g., of the classes Alphaproteobacteria, Betaprobacteria, Gammaprobacteria (e.g., organisms of the order Enterobacterales, e.g., organisms of the family Enterobacteriaceae, e.g., of the genus Proteus, (e.g., species related to Proteus mirabilis and Proteus vulgaris), or, e.g., organisms of the order Alteromonadales, e.g., of the family Shewanellaceae (e.g., of the genus Shewanella, e.g., species such as Shewanella putrefaciens, Shewanella oneidensis, Shewanella sediminis, etc.)), Deltaproteobacteria), Firmicutes, Campylobacterota (e.g., of the class Campy lobacteria, e.g., of the order, campylobacterales, e.g., of the family Helicobacteraceae, e.g., of the genus Helicobacter, e.g.. organisms related to Helicobacter pylori), Myxococcota (e g., of the class Myxococcia, e.g., of the order Myxococcales, e.g.,of the family Myxococcaceae, e.g., a species such as Myxococcus Xanthus) or Actinobacteria. Urease producing Firmicutes include, e.g., organism of the family Bacillaceae (e.g., Alkalibacillus, Alkalicoccus, Alkalihalobacillus, Bacillus, Halalkalibacillus, Halobacillus, Halolactibacillus, Lysinibacillus, Natribacillus, Natronobacillus, Priestia, Pseudobacillus, Pseudogracilibacillus, Pseudoneobacillus, Psychrobacillus, Thermalkalibacillus; for example, species such as Lysinibacillus sphaericus, Bacillus sphaericus, Bacillus subtilis, Bacillus megaterium, etc.) or. e.g., of the family Caryophanaceae (e g., of the genus Sporosarcina (e.g., species such as Sporosarcina pasteurii or Sporosarcina ureae), e.g., of the genus Bhargavaea (e.g., Bhargavaea beij ingensis)), or, e.g., of the family Paenibacillaceae (e.g., of the genus Paenibacillus)). Methods described herein may involve a consortia or coculture of two or more biological organisms, e.g., a consortia or co-culture of related or unrelated mineralizing organisms, e.g. multiple species in co-culture or multiple strains of the same species in co-culture. Methods described herein may involve a combination of one kind of enzyme (e.g., urease or carbonic anhydrase) and a microorganism that produces another enzyme (e.g., urease or carbonic anhydrase). The mineralizing microorganism may be tolerant of high osmolarity (e.g., from 4 to 30 Osm / L, e.g., from 4 to 12 Osm / L). The mineralizing microorganism may be acid tolerant. The mineralizing microorganism may be base tolerant. The mineralizing microorganism may be tolerant of a broad pH range (e.g., from acidic to basic, e.g., from about pH 2 to about pH 10). The mineralizing microorganism may have a higher acidity, basicity, ionic strength, and / or osmolarity tolerance in a system or process described herein, e.g., in solid state bioprocesses (e.g., germination, activation, expansion, proliferation, fermentation, etc.), than in corresponding liquid-medium bioprocesses.

[0161] The disclosed systems and methods may also include the use of microorganisms that are acid producing microorganisms. The acid producing microorganisms may be from the domains of archaea (e.g., haloarchaea), bacteria, or eukarya (e.g., fungi (e.g., yeasts (e.g., ofthe genus saccharomyces) or filamentous fungi (e g., of the genus Aspergillus)), and some algae (e.g., Thraustochytrids)). The acid-producing microorganism may be selected from the group consisting of: Variovorax, Klebsiella, Pseudomonas, Bacillus, Exiguobacterium, Microbacterium, Curtobacterium, Rathayibacter. Streptomyces, Raoultella, B. pumilus, B. safanensis, B. simplex, B. licheniformis, Lysinibacillus sphaericus and combinations thereof. The acid producing microorganism may be tolerant of high osmolarity. The mineralizing microorganism may be acid tolerant. The mineralizing microorganism may bebase tolerant. The mineralizing microorganism may be tolerant of a broad pH range (e.g., from acidic to basic, e.g., from about pH 2 to about pH 10). The mineralizing microorganism may have a higher acidity, basicity, ionic strength, and / or osmolarity tolerance in a system or composition or process described herein, e.g., in solid state bioprocesses (e.g., germination, activation, expansion, proliferation, fermentation, etc.), than in corresponding liquid-medium bioprocesses.

[0162] In some embodiments, the construction material may further include a binding agent. The binding agent that can be generated through a biological mechanism (such as an enzymatic process) which adheres to or encapsulates aggregate particles. In some embodiments, the binding agent may be a biological organism or enzyme. The biological organism may be any mineralizing microorganism, e.g., any urease-producing organism, acid-producing organism, or a carbonic anhydrase-producing organism. The enzyme may be urease or carbonic anhydrase. In some embodiments, the carbonic anhydrase-producing microorganisms may be the same organism as the urease-producing microorganism or the acid-producing microorganism. In some embodiments the carbonic anhydrase-producing microorganism is a different microorganism to the urease-producing microorganism.

[0163] In some cases, the processes for manufacturing a construction material disclosed herein may utilize carbonic anhydrase to improve carbon capture within the construction material and to catalyze the interconversion between carbon dioxide and water and the disassociated ions of carbonic acid. In one example, a construction material is manufactured or produced by arranging the construction material within a sealed tank with carbon dioxide and carbonic anhydrase. Magnesium hydroxide may also be utilized to improve carbon capture within the construction material. In one example, a construction material is manufactured or produced by arranging the construction material within a sealed tank with carbon dioxide and magnesium hydroxide.

[0164] In some embodiments, the process of producing a construction material may be a hydroponic process. In some embodiments, during the hydroponics process, each of a plurality of pressed aggregate units is fed with calcium ion source chemical and carbonate ion source chemical to form bio-cement (or calcium carbonate formed from biologically generated calcium ion and / or carbonate ion), which is a biologically formed calcium carbonate. In some embodiments, this biocement (calcium carbonate) formation process may happen in the voids of the pressed aggregate units (e.g., in the pore volume). In some embodiments, the calcium ion is formed biologically. In some embodiments, the carbonateion is formed biologically. In some embodiments, both the calcium ion and the carbonate ion are formed biologically. As used herein, the term “formed biologically" generally refers to the formation of a product from a starting material by a biological agent. In some embodiments, the biological agent may be one or more enzy mes, one or more microorganisms, or a combination thereof. In some embodiments, this bio-cement (calcium carbonate) formation process may bridge the gaps between aggregate particles within each of the plurality of pressed aggregate unit. In some embodiments, the hydroponic process includes some, or all of, steps of aggregate blend preparation, germination, press, drying, feeding, deodorization, and finish. In some embodiments, each of the steps of aggregate blend preparation, germination, activation, incubation, fermentation, press, dry ing, feeding, deodorization, and finish is performed with controlled parameters. In some embodiments, the product formed may be evaluated based on parameters including, but not limited to, compression strength, flexural strength, absorption, and sustaining freeze thaw cycles.

[0165] In some embodiments, the duty cycle includes a feed process that provides reagents to the framework. In some embodiments, the feed process includes a duty7cycle comprising a plurality of cycles with parameters comprising independent feed-on time and fed-off time for each cycle, and a total number of cycles. In some embodiments, the duty cycle of the feed process includes a first bath and a plurality7of subsequent baths. In some embodiments, the first bath includes a first feed-on time that is from about 80 seconds to about 120 seconds. In some embodiments, the first feed-on time is from about 80 seconds to about 85 seconds, from about 85 seconds to about 90 seconds, from about 90 seconds to about 95 seconds, from about 95 seconds to about 100 seconds, from about 100 seconds to about 105 seconds, from about 105 seconds to about 110 seconds, from about 110 seconds to about 115 seconds, or from about 115 seconds to about 120 seconds. In some embodiments, the first feed-on time is about 80, 85, 90, 95, 100, 105, 110. 115, or 120 seconds. In some embodiments, the first feed-on time is about 100 seconds. In some embodiments, the first bath includes a first feed-off time that is from about 800 seconds to about 1,000 seconds. In some embodiments, the first feed-off time is from about 800 seconds to about 820 seconds, from about 820 seconds to about 840 seconds, from about 840 seconds to about 860 seconds, from about 860 seconds to about 880 seconds, from about 880 seconds to about 900 seconds, from about 900 seconds to about 920 seconds, from about 920 seconds to about 940 seconds, from about 940 seconds to about 960 seconds, from about 960 seconds to about 980 seconds, or from about 980 seconds to about 1,000 seconds. Insome embodiments, the first feed-on time is about 800, 810, 820, 830, 840, 850, 860, 870, 880. 890, 900, 910, 920. 930, 940, 950, 960. 970, 980, 990, or 1,000 seconds. In some embodiments, the first feed-off time is about 900 seconds.

[0166] In some embodiments, each of the plurality of subsequent baths independently includes a subsequent feed-on time that is from about 80 seconds to about 120 seconds. In some embodiments, the subsequent feed-on time is independently from about 80 seconds to about 85 seconds, from about 85 seconds to about 90 seconds, from about 90 seconds to about 95 seconds, from about 95 seconds to about 100 seconds, from about 100 seconds to about 105 seconds, from about 105 seconds to about 1 10 seconds, from about 110 seconds to about 115 seconds, or from about 115 seconds to about 120 seconds. In some embodiments, the subsequent feed-on time is independently about 80, 85, 90, 95, 100, 105, 110. 115. or 120 seconds. In some embodiments, the subsequent feed-on time is independently about 100 seconds. In some embodiments, each of the plurality of subsequent baths independently includes a subsequent feed-off time that is from about 400 seconds to about 600 seconds. In some embodiments, the subsequent feed-off time is independently from about 400 seconds to about 420 seconds, from about 420 seconds to about 440 seconds, from about 440 seconds to about 460 seconds, from about 460 seconds to about 480 seconds, from about 480 seconds to about 500 seconds, from about 500 seconds to about 520 seconds, from about 520 seconds to about 540 seconds, from about 540 seconds to about 560 seconds, from about 560 seconds to about 580 seconds, or from about 580 seconds to about 600 seconds. In some embodiments, the subsequent feed-on time is independently about 400, 410, 420, 430, 440, 450, 460, 470, 480, 490, 500, 510, 520, 530, 540, 550, 560, 570, 580, 590, or 600 seconds. In some embodiments, the subsequent feed-off time is independently about 500 seconds. In some embodiments, the total number of cycles is from about 15 to about 40 cycles. In some embodiments, the total number of cycles is from about 15 to about 22, from about 17 to about 24, from about 19 to about 26, from about 21 to about 28. from about 23 to about 30, from about 25 to about 32, from about 27 to about 34, from about 29 to about 36, from about 31 to about 38, or from about 33 to about 40 cycles. In some embodiments, the total number of cycles is from about 23 to about 30 cycles. In some embodiments, the total number of cycles is about 15, 16, 17, 18. 19. 20. 21. 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40 cycles.

[0167] A temperature, humidity, and / or pH of a framed plurality of aggregate particles can be monitored and controlled during the reaction that forms the construction material.The act of forming bridging calcium carbonate crystals between at least two members of the framed plurality’ of aggregate particles may include at least one biological organism or enzyme further comprising a mineralizing enzyme (e.g., urease) or cells of a mineralizing (e.g., urease-producing or carbonic anhydrase-producing) microorganism. The act of forming bridging calcium carbonate cry stals between at least two members of the framed plurality of aggregate particles may include an acid-producing enzyme or cells of an acidproducing microorganism and / or carbonic anhydrase or cells of a carbonic anhydraseproducing microorganism. In some embodiments, the carbonic anhydrase-producing microorganisms may be the same organism as the urease-producing microorganism or the acid-producing microorganism. In some embodiments the carbonic anhydrase-producing microorganism is a different microorganism to the urease-producing microorganism. In some embodiments, a feed solution of a system or method described herein may include carbonic anhydrase enzyme and may be further fed gaseous carbon dioxide, e.g., captured carbon dioxide.

[0168] Mineralizing (e.g., urease-producing and / or carbonic anhydrase producing) microorganisms can include but are not limited to organisms in the domains of archaea (e.g., haloarchaea), bacteria (e.g., Cyanobacteriota (e.g.. cyanobacteria), Pseudomonadota, Gammaprobacteria, Firmicutes, Campylobacterota, Myxococcota), or eukarya (e.g., fungi (e.g., yeasts (e.g., of the genus saccharomyces) or filamentous fungi (e.g., of the genus Aspergillus)), protists (e.g., diatoms or coccolithophores), and some algae (e.g., Thraustochytrids)). Urease-producing and / or carbonic anhydrase-producing bacteria include organisms in the phyla Pseudomonadota (e.g., of the classes Alphaproteobacteria, Betaprobacteria, Gammaprobacteria (e.g., organisms of the order Enterobacterales, e.g., organisms of the family Enterobacteriaceae, e.g., of the genus Proteus (e.g., species such as, or related to, Proteus mirabilis and Proteus vulgaris), or, e.g., organisms of the order Alteromonadales. e.g., of the family Shewanellaceae (e.g., of the genus Shewanella, e.g., species such as, or related to, Shewanella putrefaciens, Shewanella oneidensis, Shewanella sediminis, etc.)), or Deltaproteobacteria), Firmicutes, Campylobacterota (e.g., of the class Campylobacteria, e.g., of the order, campylobacterales, e.g., of the family Helicobacteraceae, e.g., of the genus Helicobacter, e.g., organisms such as, or related to, related to Helicobacter pylori), Myxococcota (e.g., of the class Myxococcia, e.g., of the order Myxococcales, e.g., of the family Myxococcaceae, e.g., a species such as Myxococcus Xanthus), or Actinobacteria. Urease producing and carbonic anhydrase-producingFirmicutes include, e.g., organisms of the family Bacillaceae (e.g., Alkalibacillus, Alkalicoccus, Alkalihalobacillus. Bacillus. Halalkalibacillus, Halobacillus, Halolactibacillus, Lysinibacillus, Natribacillus, Natronobacillus, Priestia, Pseudobacillus, Pseudogracilibacillus, Pseudoneobacillus, Psychrobacillus, Thermalkalibacillus; for example, species such as Lysinibacillus sphaericus, Bacillus sphaericus, Bacillus subtilis, Bacillus megaterium, etc.) or. e.g., of the family Caryophanaceae (e.g., of the genus Sporosarcina (e.g., species such as Sporosarcina pasteurii or Sporosarcina ureae) . e.g., of the genus Bhargavaea (e.g., Bhargavaea beij ingensis)), or, e.g., of the family Paenibacillaceae (e.g., of the genus Paenibacillus).

[0169] Biocementation reagents can further include nutrients which promote the growth or enzy matic activities of microorganisms. In some embodiments, the nutrients include one or more of salts, amino acids, proteins, peptides, carbohydrates, saccharides, polysaccharides, fatty acids, oil, vitamins, and minerals.

[0170] Figure 1A depicts one embodiment of a biostructure manufacturing system 100. The biostructure manufacturing system 100 may produce or manufacture various biostructures including tiles, pavers, and bricks that include biocement compositions. The biostructure manufacturing system 100 includes mixing tank 102 and various subsystems including a green biostructure forming system 104. incubation system 106, and feed system 108. In some cases, the mixing tank 102 may be used to combine a plurality of aggregate particles, an inoculation composition including mineralizing enzymes and / or mineralizing (e.g., urease producing) microorganisms to form an aggregate mixture. In other cases, the mixing tank 102 may be used to combine a plurality' of aggregate particles to form an aggregate mixture, while an inoculation composition is applied subsequent to formation of the aggregate mixture into a shaped structure, such as a structure with a tile or brick shape. As depicted, tiles 132-134 may be processed and then transferred from one subsystem to another via a transportation system, such as the conveyor system 120. Although the conveyor system is represented by a belt or chain-type system in the figure, other conveyance structures (e.g., robotic pallet-mover arms, cranes, etc.) are also considered. Further, different conveyor systems may move the tiles from one stage to the next.

[0171] In some embodiments, pallets or trays provide structural support for various biostructures (e.g., tiles, pavers, and bricks) during manufacturing. The pallets or trays may support biostructures during various manufacturing phases and as the biostructures are moved within a manufacturing facility during manufacturing. In one example, themanufactured biostructures may be supported by a pallet during pressing and / or feeding phases. In some cases, the pallet may comprise a porous pallet (e.g., with holes (e.g., 1-1000 holes per pallet, or 1-10,000 holes per square meter, e.g., with holes of 1-100 mm in diameter, e.g., about 5, 10, 20, 50, or 100 mm in diameter). In one example, the holes through a porous pallet are uniformly spaced. In another example, the holes through a porous pallet are unevenly distributed across the face of the pallet (e.g., with a higher density of holes in locations underneath where biostructures will be placed). In some cases, the pallet may comprise a non-porous pallet (e.g., flat non-porous pallets or non-porous pallets with one or more depressions (e.g., bumps, undulations, channels, hollows, etc., with one or more channels (e.g., 1-1000 channels, e.g., 1-1000 channels per meter or length or width, e.g., channels of 1-100 mm in depth)). A non-porous pallet with features (e.g., channels) may provide, in certain feeding regimes, a compromise between the porous and flat non-porous pallet designs.

[0172] Figure IB depicts another embodiment of a biostructure manufacturing system 100. As depicted, the biostructure manufacturing system 100 includes mixing tank 102 and various subsystems including a green biostructure forming system 104, incubation system 106. feed system 108, reinoculation and feed system 110. tile curing system 112, and tile finishing system 114. The mixing tank 102 may be used to mix or combine a plurality of aggregate particles, an inoculation composition including mineralizing enzymes and / or mineralizing (e.g., urease producing) microorganisms to form an aggregate mixture. The system may further include an incubation system 106 for performing solid state germination, activation, expansion, proliferation, and / or fermentation in the unformed aggregate mixture or in the formed biostructure.

[0173] The green biostructure forming system 104 may form a shaped structure, such as tile 132, using a press. Although Figure IB depicts that the manufactured biostructures are tiles, other biostructures such as bricks may also be manufactured. The incubation system 106 may apply an inoculation solution to a tile, such as tile 133, and may provide a climate-controlled environment for growing or increasing the mineralizing (e.g., urease producing or carbonic anhydrase producing) microorganisms within a shaped structure.

[0174] The feed system 108 may include a fluid delivery system for applying a feed solution to a tile, such as tile 134. The reinoculation and feed system 110 may reapply an inoculation solution and / or one or more feed solutions to a tile, such as tile 135. The tile curing system 112 may provide an environment for curing a tile, such as tile 136. In oneexample, the tile curing system 112 may provide an enclosed or partially enclosed climate- controlled environment at a particular temperature and humidity for curing of tiles. The tile finishing system 114 may apply a sealant to surfaces of a tile, such as tile 137, or perform polishing of the tile.

[0175] The biostructure manufacturing system 100 may be used to manufacture tiles, such as tiles 132-137, during different phases of construction. The tiles 132-137 may be transported between different subsystems via a conveyor system 120. The conveyor system 120 may include a conveyor belt and automated cranes for transporting the tiles 132-137 between the different subsystems. In some cases, the conveyor system 120 may include one or more conveyor belts and / or one or more overhead cranes. One or more of the subsystems including biostructure forming system 104, incubation system 106, feed system 108, reinoculation and feed system 110, tile curing system 112, and tile finishing system 114 may reside within a climate-controlled environment that regulates the ambient temperature and humidity for tiles (or other biostructures) being processed by the climate-controlled environment.

[0176] Figure 1C depicts one embodiment of a portion of a biostructure manufacturing system that includes a climate controlled environment 122 for regulating the ambient temperature and humidity of biostructures within the climate controlled environment 122, such as tiles 134 and 135. The conveyor system 120 may transport tiles, such as tile 132. into and throughout the climate controlled environment 122. As depicted, the climate controlled environment 122 includes gas sensors 105, temperature and humidity sensors 109. and computing system 101 all in communication with each other. The gas sensors 105 may monitor gas levels for a particular gas or detect an amount of a gas that is present within the climate controlled environment 122. As examples, the gas being sensed may include carbon dioxide gas or ammonia gas. The temperature and humidity sensors 109 monitor the temperature and humidity within the climate controlled environment 122. In some cases, the climate controlled environment 122 may include temperature and humidity equipment that regulates the temperature and humidity of the climate controlled environment 122 using temperature regulated air flow. The computing system 101 may take as inputs the temperature and humidity7of the climate controlled environment 122 and dynamically configure manufacturing parameters for the feed system 108. As examples, the manufacturing parameters for the feed system 108 may include an average feed rate forapplying a feed solution to a biostructure, the minimum feed rate for applying a feed solution to a biostructure, and the maximum feed rate for applying a feed solution to a biostructure.

[0177] In some cases, the feed system 108 may apply a feed solution to one or more biostructures (e.g., tiles) concurrently. The feed system 108 may apply a first feed solution at a first feed rate to the tile 134 and a second feed solution at a second feed rate different from the first feed rate to the tile 135. A feed rate profile may correspond to a feed rate waveform over a period of time or to the feed rates used to apply a feed solution to a biostructure over the period of time. The feed rate waveform may correspond to one or more feed pulses. In one embodiment, the feed system 108 may apply a first feed solution to tile 134 using a first feed rate profile (e.g., with a first feed rate) and a second feed solution to tile 135 using a second feed rate profile (e.g., with a second feed rate less than the first feed rate).

[0178] Figure ID depicts one embodiment of a portion of a biostructure manufacturing system that includes a climate controlled environment 122 that includes a crust formation system 107 and a feed system 108. In some embodiments, the gas sensor 105 may monitor and detect ammonia gas levels and / or carbon dioxide levels within the climate controlled environment 122. The computing system 101 may dynamically configure the crust formation system 107 to apply localized heat and blown air for a specified period of time to cause a surface portion of a biostructure, such as tile 132. to harden and form a structural crust on an external surface of the biostructure. The thickness of the structural crust formed for the biostructure may be increased by increasing the period of time that the localized heat and blown air is blown over the surface of the biostructure. The thickness of the structural crust formed for the biostructure may also be adjusted by regulating the ambient temperature and humidity within the climate controlled environment 122 while the localized heat and air is applied to the biostructure. A maximum thickness of the structural crust may be set to ensure that a feed solution may penetrate the structural crust while the structural crust provides structural support for the biostructure during subsequent applications of a feed solution. A minimum thickness of the structural crust may be set to ensure that a green biostructure may withstand an initial feed cycle without significant deformation. In some embodiments, the crust formation system 107 and the feed system are in separate climate controlled environments 122, each with its own gas sensors 105. temperature and humidity sensors 109 and computing system. Biostructures may be moved between the two climate controlled environments by a linked conveyor system 120.

[0179] The gas sensors 105 and temperature and humidity sensors 109 may include monitoring or sensing circuitry or sensing equipment.

[0180] Figure IE depicts one embodiment of a portion of a biostructure manufacturing system that includes a multi-pass feed system 126 that may apply a recirculated feed solution to a biostructure, such as tile 134. Some tiles within a biostructure manufacturing system may be fed using a single-feed process while other tiles may be fed using a multi-pass feed system. After a feed solution has been applied to a biostructure, such as tile 134, the residual feed solution 139 that is not absorbed by the biostructure may be collected using a feed collection tank 128. The pH sensor 124 may determine the pH of the residual feed solution 139. The level sensor 123 may determine the amount of liquid within the feed collection tank 128 and may determine the amount of residual feed solution 139 that has been collected. A pump 127 may be used to recirculate a portion of the residual feed solution 139 to be used during subsequent feed cycles. In one embodiment, the multi-pass feed system 126 may be reused a feed solution over five different feed cycles. In some embodiments, the multi-pass feed system 126 includes an acid supply to periodically reduce the pH of the feed solution. The acid supply system may deliver an acid directly to the multi-pass feed system 126 (e.g.. between pump 127 and multi-pass feed system 126), or to the feed collection tank 128, or directly to tile 134 (e.g., via a separate spray or drip nozzle).

[0181] Figure IF depicts one embodiment of a portion of a biostructure manufacturing system that includes a feed system 108 that applies a single-use feed solution to a biostructure, such as tile 144, that is not recirculated. The feed system 108 may apply a feed solution using a single pass approach wherein the feed solution applied to a biostructure is not recirculated nor reapplied to the biostructure. Residual feed solution that is not absorbed by the biostructure during deposition, or is pushed out by a subsequent feed cycle, is collected in the feed collection tank 129. The pH of the residual feed solution is monitored by the pH sensor 124. The volume of liquid within the feed collection tank 129 is monitored by the level sensor 123. The level sensor 123 may be used to determine a residual flow rate for the residual feed solution being collected by the feed collection tank 129. The biostructure, such as tile 144, may be positioned within a formwork 152 that includes holes 153 that may promote drainage of the applied feed solution and liquid flow through the biostructure.

[0182] In some cases, the pH of the residual feed solution and the residual flow rate may be used to set biostructure manufacturing parameters including feed rate, feed pressure, feedmolarity, feed rate profile, feed pH level, temperature, and humidity during subsequent feed cycles. The feed system 108 may be used to apply multiple single-use or single-pass feed cycles to a biostructure, wherein the feed solution applied to the biostructure is not reused or reapplied to the biostructure. In some cases, the feed collection tank 129 may instead, or in addition, include an electrical conductivity (EC) detector 125, to monitor change in electrical conductivity of the residual feed solution. In some cases, the EC of the residual feed solution and the residual flow rate may be used to set biostructure manufacturing parameters including feed rate, feed pressure, feed molarity', feed rate profile, feed pH level, temperature, and humidity during subsequent feed cycles.

[0183] Figure 1G depicts one embodiment of a portion of a biostructure manufacturing system that includes a feed system 108 that applies a single-pass feed solution to a biostructure, such as tile 144. As depicted, the tile 144 has been positioned within a formwork 157 that allows the tile 144 to be completely submerged in feed solution 159. The top of the conveyor system 120 may prevent the applied feed solution from being discharged through holes in the formwork 157 while the tile 144 is completely submerged in the feed solution 159. As the tile 144 is moved along the conveyor system 120, the holes may become uncovered allowing the feed solution 159 to drain from the formwork 157 and allow the tile 144 to dry out.

[0184] Figures 1H-1K depict various embodiments of a portion of a feed system 108 with one or more spray nozzles in a nozzle configuration for applying a feed solution to a biostructure. As depicted in Figure 1H, a spray nozzle 180 is configured to apply a feed solution onto a surface of a biostructure. The nozzle 180 may provide a spray pattern within the spray cone 181 to generate a liquid distribution 188 on the surface of the tile 144. With the spray pattern depicted in Figure 1H, the liquid distribution 188 does not uniformly coat the entire surface of the tile 144. In some cases, an increase in the feed rate or feed pressure may cause the spray cone 181 to enlarge and allow the entire surface of the tile 144 to be non-uniformly coated with a feed solution.

[0185] In some embodiments, the feed solution may be applied to a surface of a biostructure using a nozzle, a control valve, and / or restrictive tubing. The control valve may include an actuated valve, an occlusive valve, a blocking valve, a compressive valve, or a constricting valve.

[0186] As depicted in Figure II, two spray nozzles 180 and 182 are used to spray two spray patterns corresponding to spray cones 181 and 183. The two spray patterns form liquid distribution 189 on the surface of the tile 144. The orientation and feed pressures applied to the nozzles 180 and 182 have created a spray gap 186 in which a feed solution is not applied across the entire surface of the tile 144 and a gap in the liquid distribution 189 has formed in which the feed solution is not applied to a middle portion of the surface of the tile 144. As depicted, a nozzle pressure applied to the nozzles 180 and 182 may be insufficient to fully apply the feed solution to the entire surface of the tile 144 and therefore a spray gap 186 may exist between the spray cones 181 and 183 in which the feed solution is not directly applied to the surface of the tile 144. Setting a minimum flow rate or flow pressure for the feed solution may allow the spray gap 186 to be eliminated such that the feed solution is applied to the entire surface of the tile 144.

[0187] As depicted in Figure 1J. the orientation and nozzle pressure for the two spray nozzles 180 and 182 have created a spray overlap 187 in which the spray cones 181 and 183 intersect or overlap. The orientation and nozzle pressure for the two spray nozzles 180 and 182 allow a feed solution to be applied more uniformly on the surface of the tile 144. The liquid distribution 190 of the feed solution applied to the surface of the tile 144 is more uniform than the liquid distribution 189 in Figure II. However, some of the feed solution sprayed by the spray nozzles 180 and 182 is wasted or is not sprayed onto the top surface of the tile 144.

[0188] Figure IK depicts one embodiment of a portion of a feed system 108 with spray nozzles 180 and 182 with orientations and nozzle pressures that form a nearly uniform liquid distribution 190 on the surfaces of multiple biostructures that include tiles 144-146. In some embodiments, the shape of the nozzle openings may be triangular, such that the part of the spray cone that impacts the tile where the distance between the nozzle and the tile has a wider footprint on the tile than the part of the spray cone that impacts the tile furthest from the nozzle, thus the impact pressure of the spray may also be nearly equalized across the width of the tile. An advantage of such a system is to provide an even distribution of fluid in a way that minimizes surface damage to the green tile surface.

[0189] In some cases, for a spray feeding system, impact pressure for a fluid has a greater impact than nozzle shape or flow rate. One technical issue is that as shapes may not have sufficiently hardened and will harden over time, the impact pressure may have to be limited until a construction material is sufficiently hardened. In one embodiment, the impactpressure for the fluid is set to a first pressure during an initial time period and then increased to a second pressure greater than the first pressure after the initial time period has lapsed. In some cases, the impact pressure is a function of the hardness of the construction material during the manufacturing process. The impact pressure may be increased and / or decreased over time during the manufacturing process as the construction material hardens.

[0190] Figure IL depicts a portion of a feed system that includes nozzles 180 and 182 for applying a feed solution to biostructures including tiles 144 and 146. A sensor 145 may include an IR sensor or an image sensor that is used to detect when a tile or other biostructure is within a spray cone or spray pattern of the nozzles 180 and 182. The feed system may only spray the feed solution when a biostructure is within a spray pattern for the nozzles 180 and 182 as the sensor 145 may be used to detect the presence of the biostructure within the spray pattern. The application of the feed solution may be turned off in order to conserve the feed solution.

[0191] In some cases, the nozzles 180 and 182 may have different nozzle shapes or apply different spray patterns. The nozzles 180 and 182 and the nozzle pressures applied while the feed solution is applied to a biostructure may modulate the deposition of the feed solution to a surface of the biostructure. In one example, the spray patterns may project a pyramid e.g., a rectangular base, a square base, or a cone with circular base, or an ellipsoidal base onto the surface of the biostructure.

[0192] Figure IM depicts one embodiment of various components of the computing system 101. The components within the computing system 110 may include real hardware computing devices or virtual computing devices, such as one or more virtual machines. As depicted, the computing system 101 includes hardware-level components and softwarelevel components. The hardware-level components may include one or more processors 170, one or more memories 171, and one or more disks 172. The one or more processors170 may include one or more processing units, such as one or more CPUs, one or more GPUs, and / or one or more NPUs. The one or more memories 171 may include one or more types of memory (e.g., RAM, SRAM, DRAM, EEPROM, Flash). The one or more disks 172 may include a hard disk drive and / or a solid-state drive. Both the one or more memories171 and the one or more disks 172 may include hardware storage devices. A storage device may correspond to the one or more memories 171 or the one or more disks 172.

[0193] The software-level components may include software applications and computer programs. The pre-feed crust formation application 191. conveyor control application 193, environmental control application 194, impulse control application 195, regeneration control application 196, and feeding control application 192 may be stored or implemented using software or a combination of hardware and software. In some cases, the softwarelevel components are run using a dedicated hardware server. In other cases, the softwarelevel components may be run using a virtual machine or containerized environment running on a plurality of machines. In various embodiments, the software-level components may be run from the cloud (e.g., the software-level components may be deployed using a cloudbased compute and storage infrastructure). Applications such as those in Figure IM may include sub-applications, for example, the environmental control application may contain a temperature control application, a humidity control application 194, an air-flow control application, etc. In another example, the conveyor control may include control applications for multiple conveyor systems (e.g., in-bath conveyor application(s), inter-system conveyor application(s), intra-system conveyor application(s), feed conveyor application(s). etc. In another example, the feeding control application may include a pressure control application, an aperture control application, a flow control application, a temperature control application, a source control application, etc.

[0194] As depicted in Figure IM, the software-level components may also include virtualization layer processes, such as virtual machine 173, hypervisor 174, container engine 175. and host operating system 176. The hypervisor 174 may include a native hypervisor (or bare-metal hypervisor) or a hosted hypervisor (or type 2 hypervisor). The hypervisor 174 may provide a virtual operating platform for running one or more virtual machines, such as virtual machine 173. A hypervisor may include software that creates and runs virtual machine instances. Virtual machine 173 may include a plurality of virtual hardware devices, such as a virtual processor, a virtual memory, and a virtual disk. The virtual machine 173 may include a guest operating system that has the capability to run one or more software applications. The virtual machine 173 may run the host operation system 176 upon which the container engine 175 may run.

[0195] The container engine 175 may run on top of the host operating system 176 in order to run multiple isolated instances (or containers) on the same operating system kernel of the host operating system 176. Containers may facilitate virtualization at the operating system level and may provide a virtualized environment for running applications and theirdependencies. Containerized applications may include applications that run within an isolated runtime environment (or container). The container engine 175 may acquire a container image and convert the container image into running processes. In some cases, the container engine 175 may group containers that make up an application into logical units (or pods). A pod may contain one or more containers and all containers in a pod may run on the same node in a cluster. Each pod may serve as a deployment unit for the cluster. Each pod may run a single instance of an application.

[0196] In some embodiments, the depicted components of the computing system 101 including the pre-feed crust formation application 191, conveyor control application 193, environmental control application 194, impulse control application 195, regeneration control application 196, and / or the feeding control application 192 are implemented in the cloud or in a virtualized environment that allows virtual hardware to be created and decoupled from the underlying physical hardware. The pre-feed crust formation application 191 may configure one or more manufacturing parameters for the crust formation system 107 and control the operation of the crust formation system 107 to apply localized heat and blown air to tiles being manufactured. The feeding control application 192 may configure one or more manufacturing parameters for the feed system 108 and control the operation of the feed system 108 to deliver or apply a feed solution to a shaped structure (e.g., atile) over one or more feed cycles. The conveyor control application 193 may configure one or more manufacturing parameters for the conveyor system(s) 120 (or one or more sub-systems, such as the in-bath conveyor system (3102) and / or impulse delivery system 3103 or inter-system conveyor system 3104) and control the operation of the conveyor system(s) 120 to, e.g., transfer structures between systems (e.g., between incubation system 106 and feeding system 108, between the feeding system 108 and the curing system 112, between the feeding system 108 and the finishing system(s) 114 / 3006, between the press 104 and the incubation system 106 or feeding system 108, etc.), through systems (e.g., the feeding system 108), e g., causing one or more actuators, robotic arms, belts, cranes, conveyor belts, etc., to move. The environmental control application may configure one or more manufacturing parameters in the feeding system 108 or incubation system 106 to maintain a temperature and humidity within a determined range, or to adjust the temperature or humidity in response to information from one or more sensors. The impulse control application may configure one or more manufacturing parameters for the conveyor system(s) 120 (e.g., the in-bath conveyor 3102 and / or impulse delivery system 3103 (e.g., a piston, a plow, a diaphragm, ascrew, or a blade)) or another system of the feed system 108 (e.g., pump(s) 3012, mixer(s) 3016,) and control the operation of the conveyor system(s) 120 to, e.g.. causing one or more actuators, robotic arms, belts, cranes, conveyor belts, etc., and / or control one or more pump(s) 3012, mixer(s) 3016, pistons, plows, diaphragms, screws, or blades, to move. The regeneration control application may configure one or more manufacturing parameters for the regeneration system

[0197] Figure IN depicts one embodiment of a biostructure manufacturing system 198 that includes a nozzle 180 for applying a feed solution onto a surface of a biostructure, such as tile 144. An effluent collection tank 199 may correspond to the feed collection tank 129 in Figure IF (which may both be components of an effluent system 3008 of Figures 30A, 30C, 30D) and be used to collect residual feed solution that was not absorbed by the biostructure. The biostructure manufacturing system 198 includes a Post Run Clean in Place (CIP) tank, which may include a tank for storing a secondary fluid, such as water for maintaining moisture content within biostructures between feed cycles or for cleaning the lines of the feeding system after a run. The biostructure manufacturing system 198 includes effluent monitoring equipment for monitoring the pH and electrical conductivity of the effluent within the effluent collection tank 199. The biostructure manufacturing system 198 may include a feed temperature regulator for heating or cooling a feed solution and to maintain a temperature of the feed solution to be between 18 and 42 degrees Celsius, e.g., between, 25 and 40 degrees Celsius, e.g., between 28 and 34 degrees (e.g., about 30 degrees) or, e.g., between 34 and 38 degrees Celsius, e.g., between 35 and 36 Celsius (e.g.. about 35 degrees Celsius). An effluent tank may include an element to inactivate mineralizing enzymes or microorganisms (e.g., as part of a biologies inactivation system, such as shown in Figures 30D and 321), for example, a heating element, a UV light source, a source of ozone gas, etc. Alternatively, the element that inactivates mineralizing enzymes or microorganisms may be upstream or downstream of the effluent tank. In some embodiments of systems and processes described herein, sterilized or pasteurized effluent may be re-used as feed solution (e.g., pumped back into a feed reservoir or bath with added cementation reagents).

[0198] Figure 10 depicts another embodiment of a biostructure manufacturing system 100. As depicted, the biostructure manufacturing system 100 includes mixing tank 102 and various subsystems including a green biostructure forming system 104 (e.g.. a press, e.g., to form the aggregate into shaped structures, e.g., tiles, pavers, blocks, veneers, etc.),incubation system 106 (e.g., to perform solid state germination, activation, and / or expansion (e.g., proliferation) of mineralizing microorganisms and / or crust formation), feed system 108 for providing cementation reagent(s), and tile curing system 112 (e.g., for storing tiles after / between feed cycles to allow binder formation). The mixing tank 102 may be used to mix or combine a plurality of aggregate particles, mineralizing enzymes and / or microorganisms (e.g., urease-producing microorganisms) (e g., in an inoculation composition) to form an aggregate mixture. The system may further include an incubation system 106 for performing solid state germination, activation, and / or expansion (e.g., proliferation) in the unformed aggregate mixture or in the formed biostructure, which may be the same system as used for formed biostructures, or a separate system.

[0199] Figure IP depicts another embodiment of a biostructure manufacturing system 100. As depicted, the biostructure manufacturing system 100 includes mixing tank 102 and various subsystems including a green biostructure forming system 104 (e.g.. a press, e.g., to form the aggregate into shaped structures, e.g., tiles, pavers, blocks, veneers, etc.), incubation system 106 (e.g., to perform solid state germination, activation, and / or expansion (e.g., proliferation) of mineralizing microorganisms and / or crust formation), feed system 108 for providing cementation reagent(s), and tile finishing system 114 (e.g., for rinsing, drying, honing, polishing, etc.). In this embodiment, there is no separate curing system, as tiles cure in the feeding system (e.g., in a feed bath, e.g., as depicted in Figures 31A-I, or, e.g., in a spray or drip-based feeding system configured for continuous production).

[0200] Figure 2A depicts one embodiment of a cross-sectional view of a construction material 68. The construction material 68 may include a plurality of aggregate particles that have been formed or pressed into a shape, such as a tile shape or a brick shape. The construction material 68 has a top surface 62 and a bottom surface 64 opposite to the top surface 62. A vertical surface 66 may connect or be orthogonal to the top surface 62 and the bottom surface 64. As depicted in Figure 2A, the vertical surface 66 extends in a first direction (e.g., the Z direction), the top surface 62 extends in a second direction orthogonal to the first direction, the bottom surface 64 extends in the second direction (e.g., the X direction), and a horizontal slice 67 is cut through the construction material 68 and extends in the second direction (e.g., the X direction). The horizontal slice 67 may have a material thickness (e.g., a thickness of O. lmm to 10cm, e.g., 0.1 mm to 1 mm, 1 mm to 2 mm, 1 mm to 5 mm, 1 mm to 1 cm, e.g., 1 cm to 2 cm, 1 cm to 5 cm, or 1 cm to 10 cm, e.g., about 0. 1mm. 0.5 mm, 1 mm, 1.5 mm. 2 mm, 5 mm, 1 cm, 2 cm, or 10 cm) and thus may include a planar section that extends in the second direction.

[0201] Figure 2B depicts one embodiment of a side view of the construction material 68 shown in Figure 2A. As depicted in Figure 2B. a first planar section 54 of the construction material 68 that abuts the top surface 62 has a first thickness 72 and a second planar section 55 of the construction material 68 that abuts the bottom surface 64 has a second thickness 76. The first thickness 72 may be substantially the same as the second thickness 76. In one example, the first thickness 72 may include 1.2cm and the second thickness 76 may include 1.1cm. A third thickness 74 of a middle planar section of the construction material 68 arranged between the first planar section and the second planar section may be determined based on a height of the construction material 68 minus the first thickness 72 and the second thickness 74. The height of the construction material 68 may correspond to the height of the vertical surface 66 in Figure 2A. The vertical surface 66 of the construction material 68 may include a vertical wall that connects the top surface 62 to the bottom surface 64.

[0202] Referring to the construction material 68 depicted in Figures 2A-2B, the top surface 62 may be parallel with the bottom surface 64. The top surface 62 may be substantially opposite the bottom surface 64. The vertical surface 66 may include one of a number of vertical walls (or walls) that are between the top surface 62 and the bottom surface 64. The vertical surface 66 may structurally connect (or connect) the top surface 62 and the bottom surface 64. A material body of the construction material 68 may encompass or enclose the top surface 62, the vertical surface 66, and the bottom surface 64. The top surface 62, the vertical surface 66, and the bottom surface 64 may include outer surfaces of the material body of the construction material 68.

[0203] As depicted in Figure 2B. the construction material 68 includes a portion 52 of the construction material 68 that is enlarged in Figures 2C-2D.

[0204] Figure 2C depicts one embodiment of aggregate particles within the construction material 68 of Figure 2B prior to compression. Figure 2D depicts one embodiment of aggregate particles within the construction material 68 of Figure 2B after compression of the aggregate particles. The aggregate particles include aggregate particle 98, which may include a particle of sand or gravel. As depicted in Figures 2C-2D, the size of the pore space between the aggregate particles has reduced due to compression of the aggregate particles. Porosity may refer to the pore space volume within a planar section of the constructionmaterial 68 divided by the total volume of the planar section. The path of fluid flow 92 is less restrictive than the path of fluid flow 94 due to compression occurring within the construction material.

[0205] As depicted in Figure 2D, a biological organism 99 has produced an environment in which calcium carbonate 97 has formed to structurally connect some of the aggregate particles including aggregate particle 98.

[0206] Due to compression of the construction material, the porosity has been reduced as well as the spacing between the aggregate particles. With concrete, increased aggregate density correlates with improved compressive strength; however, increased aggregate density also reduces pore space. One technical issue with biocement products and construction materials that include biocement is that smaller pore sizes lead to less space to hold microorganisms (e.g., bacteria) and less space to hold feed for the enzy mes and / or microorganisms (e.g., bacteria). Moreover, narrower pore throats caused by smaller pore sizes may lead to reduced flow of feed through the construction material. The combination of less space to hold microorganisms (e.g. bacteria), less space to hold feed for the enzymes or microorganisms (e.g., bacteria), and reduced flow of feed may lead to less bridging calcium carbonate within the construction material and to reduced compressive strength.

[0207] Figure 2E depicts one embodiment of experimental results 82 showing the average compressive strength (in psi) for a construction material over a range of wet green densities (in g / cc) for the construction material prior to the formation of bridging calcium carbonate and / or other binding agents within the construction material. As depicted in Figure 2E, the average compressive strength of the construction material when in a finished product state varies with the wet green density of the construction material prior to application of biocementation processes. As depicted, a wet green density of 2.10 g / cc provides an average compressive strength of 180 psi, a wet green density of 1.80 provides an average compressive strength of 590 psi, and a wet green density of 1.95 provides the highest average compressive strength of 850 psi. The maximum average compressive strength of the construction material occurs when the wet green density is 1.95 g / cc. The wet green density of the construction material prior to the application of biocementation processes may have a significant impact on the resulting average compressive strength of the construction material. The construction materials tested in this experiment were fed with multiple static feed solutions, eliminating confounding variables associated with flow- through feeding.

[0208] Figure 2F depicts one embodiment of a graph showing the percentage of total pore volume vs pore diameter for a construction material. As depicted, most pores within the construction material have diameters that are between 170mm and 300mm, with a pore diameter close to 200mm being the most common pore diameter size.

[0209] Figure 2G depicts one embodiment of experimental results 83 showing the average change in electrical conductivity (or delta EC) for a construction material over a range of average dry green densities (in g / cc) for the construction material. The average change in electrical conductivity is one metric for measuring how fast a biocementation process is proceeding. The amount of cementation reagents consumed in a feed cycle varies with the green density of the construction material. As depicted, the maximum average delta EC occurs with an average dry green density of close to 1.75 g / cc. The construction materials tested in this experiment were fed with multiple static feed solutions, eliminating confounding variables associated with flow-through feeding.

[0210] Figure 2H depicts one embodiment of a graph showing the total slice porosity over horizontal slice position from a non-preferred surface of a construction material. In reference to Figure 2A, horizontal slice 67 includes a horizontal slice through the construction material 68. As depicted, horizontal slices through the construction material from 1.7mm to 1.8mm from the non-preferred surface of the construction material has a total slice porosity of about 6%.

[0211] Figure 21 depicts one embodiment of experimental results 80 showing total open pore porosity vs slice number through a construction material. Figure 2J depicts one embodiment of a construction material body 69 that includes aggregate particles including aggregate particle 98 that have been bound together using calcium carbonate including calcium carbonate 97. As depicted, a first horizontal slice 84 (Slice #1) includes the topmost slice through the construction material body 69, a second horizontal slice 85 (Slice #953) includes an internal slice through the construction material body 69, and a third horizontal slice 86 (Slice #1961) include the bottom-most slice through the construction material body 69. The total open pore porosity for the second horizontal slice 85 (Slice #953) is close to 1%. The total open pore porosity for the first horizontal slice 84 (Slice #1) and the third horizontal slice 86 (Slice #19 1 ) are greater than 2%.

[0212] Figures 3A-3B depict a flowchart describing one embodiment of a process for manufacturing a construction material. In one embodiment, the process of Figures 3A-3B isperformed using one or more subsystems of the biostructure manufacturing system 100 depicted in Figure IB. In some embodiments, the process of Figures 3A-3B is performed using a system for manufacturing a construction material that includes a climate controlled environment, such as the climate controlled environment 122 in Figures 1C-1E, and / or a feed system, such as the feed system 108 depicted in Figures 1A-1D, 10, and IP.

[0213] In step 302, a plurality of aggregate particles is acquired. The plurality' of aggregate particles may be provided from a mix of aggregate materials. The aggregate materials may include sand, gravel, and / or granular materials. In step 304. an inoculation composition (e.g., including mineralizing enzymes and / or microorganisms) is acquired. In step 306, the inoculation composition is applied to the plurality' of aggregate particles. In one example, the inoculation composition is part of an inoculation solution that is applied to the plurality of aggregate particles using an inoculation and incubation system, such as the incubation system 106 in Figure 1A. In step 308, the plurality of aggregate particles is formed into a shaped structure. In one example, the plurality of aggregate particles is formed using a press to compact the plurality' of aggregate particles or to form the plurality of aggregate particles into a desired shape for a finished biocement product. The shaped structure may correspond to a tile shape or a brick shape. The forming of the plurality’ of aggregate particles into the shaped structure may be performed using a biostructure forming system, such as the biostructure forming system 104 in Figures 1A-1B.

[0214] In step 310, a feed solution that includes a cementation reagent is acquired. The cementation reagent may be used to promote calcite precipitation to form bonds between aggregate particles within the plurality of aggregate particles. In step 312, the feed solution is set to a first temperature and / or a first pH. In step 314, the feed solution is applied to the shaped structure. In some cases, the feed solution is applied to the shaped structure during a first time period while the feed solution is at the first temperature and / or has the first pH.

[0215] In step 316, the feed solution is set to a second temperature and / or a second pH. In step 318, the feed solution is applied to the shaped structure during a second time period subsequent to the first time period while the feed solution is at the second temperature and / or has the second pH.

[0216] In step 320, environmental conditions are provided to cause a set of bridging calcium carbonate crystals to form between at least two particles of the plurality of aggregate particles while the feed solution is applied to the shaped structure. The environmentalconditions may include temperature conditions, humidity conditions, and atmospheric pressure conditions. The environmental conditions may be provided via a climate controlled environment, such as the climate controlled environment 122 in Figures 1C-1E.

[0217] In step 322. a residual pH for the feed solution is determined. In one example, the residual pH is determined using a pH sensor, such as the pH sensor 124 in Figure IF. In step 324, a curing temperature and a curing humidity is determined based on the residual pH for the feed solution. In step 326, a curing time period is determined based on the residual pH for the feed solution. In step 328, the shaped structure is cured for the curing time period while the shaped structure is within an environment at the curing temperature and the curing humidity. In step 330, a cleaning process is performed to the shaped structure subsequent to curing the shaped structure. The cleaning process may include a rinsing process, a soaking process, and / or a deodorizing process. The shaped structure may be soaked in water or rinsed with water. In some cases, an inoculation solution and / or a feed solution may be reapplied to the shaped structure subsequent to the curing of the shaped structure.

[0218] Figures 3C-3D depict a flowchart describing another embodiment of a process for manufacturing a construction material. In one embodiment, the process of Figures 3C- 3D is performed using one or more subsystems of the biostructure manufacturing system 100 depicted in Figure IB. In some embodiments, the process of Figures 3C-3D is performed using a system for manufacturing a construction material that includes a climate controlled environment, such as the climate controlled environment 122 in Figures 1C-1E, and / or a feed system, such as the feed system 108 depicted in Figures 1 A-1D, 10, and IP.

[0219] In step 342, a shaped structure that includes mineralizing (e.g., urease producing or carbonic anhydrase-producing) microorganisms and a plurality of aggregate particles is provided. The mineralizing (e.g., urease producing or carbonic anhydrase producing) microorganisms and the plurality of aggregate particles may be added to a mixing tank, such as the mixing tank 102 in Figure IB. In step 344, a feed solution that includes a cementation reagent is acquired. In step 346, a pore volume for the plurality of aggregate particles is estimated. In one example, the pore volume may be estimated based on a composition of the plurality of aggregate particles and the dimensions of the shaped structure. The composition of the plurality of aggregate particles may include two different sets of aggregate particles with different mean particle diameters. In one example, the two different sets of aggregate particles may include a first set of particles with mean particle diametersof between 1mm and 100mm and a second set of particles with mean particle diameters that are less than 1mm.

[0220] In step 348, an ambient temperature, an ambient humidity, and a pH of the feed solution are determined. In step 350. a single-use feed volume for the feed solution is set based on the pore volume for the plurality of aggregate particles, the ambient temperature, the ambient humidity, and / or the pH of the feed solution. In step 352, one or more feed parameters are set based on the ambient temperature, the ambient humidity, and / or the pH of the feed solution. In step 354, the single-use feed volume of the feed solution is applied to the shaped structure during a first set of single-use feed cycles using the one or more feed parameters.

[0221] In step 356, an amount of overspill that has occurred while applying the singleuse feed volume is determined or detected. In step 358, an amount of the feed solution that was not absorbed by the shaped structure while applying the single-use feed volume is determined. In step 360. the single-use feed volume is adjusted (e.g., increased or decreased) based on the amount of overspill and / or the amount of the feed solution that was not absorbed by the shaped structure. In step 362, the one or more feed parameters are adjusted (e.g., increased or decreased) based on the amount of overspill and / or the amount of the feed solution that was not absorbed by the shaped structure.

[0222] In step 364, the remaining single-use feed volume of the feed solution is applied to the shaped structure during a second set of single-use feed cycles using the adjusted one or more feed parameters. In step 366, a dryout condition is detected while applying the single-use feed volume. In step 368, one or more moisture cycles are applied to the shaped structure in response to detection of the dryout condition. In step 370, the ambient temperature and / or the ambient humidity are adjusted (e.g.. increased or decreased) in response to detection of the dryout condition. In step 372, a curing procedure is performed to the shaped structure subsequent to application of the single-use feed volume of the feed solution.

[0223] Figure 4A depicts one embodiment of a graphical representation of four feed cycles applied to a construction material. The four feed cycles may correspond to four feed cycles applied to tile 134 in Figure IB using feed system 108. A first feed cycle Cl is applied to the construction material providing a feed volume of V6 (e.g., 600 mL). A second feed cycle C2 is applied to the construction material subsequent to the first feed cycle providinga feed volume of V4 (e.g., 400 mL). A third feed cycle C3 is applied to the construction material subsequent to the second feed cycle providing a feed volume of V3 (e.g., 300 mL). A fourth feed cycle C4 is applied to the construction material subsequent to the third feed cycle providing a feed volume of V2 (e.g., 200 mL). In some cases, the feed volume may reduce or monotonically decrease with each successive feed cycle.

[0224] Figure 4B depicts one embodiment of a graphical representation of four feed pulses applied to a construction material. The four feed pulses may be applied to a tile, such as tile 134 in Figure IB, using a feed system, such as feed system 108 in Figure IB. As depicted, a first feed pulse occurs between times T1 and T2 with a first average feed rate (e.g., 600ml per minute), a second feed pulse occurs between times T3 and T4 with a second average feed rate (e.g., 400ml per minute), a third feed pulse occurs between times T5 and T6 with a third average feed rate (e.g.. 300ml per minute), and a fourth feed pulse occurs between times T7 and T8 with a fourth average feed rate (e.g.. 200ml per minute). In some cases, the average feed rate may reduce or monotonically decrease with each successive feed pulse or with each successive feed cycle. A feed cycle may include one or more feed pulses. As depicted in Figure 4B, the first feed pulse includes a single pulse with a pulse width corresponding to the time difference between times T1 and T2 and a pulse height equal to the average feed rate R6. Between feed pulses, a dry interval, such as dry interval 432 between times T6 and T7, may allow' the construction material to dry out between feed cycles. In one example, the time difference between times T5 and T6 may include three minutes and the time difference between times T6 and T7 may include three hours.

[0225] Figure 4C depicts one embodiment of a set of pulses corresponding to the first feed pulse between times T1 and T2 in Figure 4B. Instead of having a single pulse with a pulse width corresponding to the time difference between times T1 and T2, three feed pulses 435-437 each w ith a pulse width equal to one ninth of the time difference betw een times T1 and T2 and with a pulse height equal to three times the average feed rate R6 may be used to provide the same feed volume as the first feed pulse with a pulse width corresponding to the time difference between times T1 and T2 depicted in Figure 4B. As depicted, the feed rate of three times the average feed rate R6 (i.e., 3*R6) is between the maximum feed rate (FR) 412 and the minimum feed rate 414. The maximum feed rate 412 may be set such that a fluid pressure of feed solution does not damage a surface of the construction material to which the feed solution is being applied. The minimum feed rate 414 may be set such thata fluid pressure for applying a feed solution to a surface of the construction material allows the feed solution to be applied to the entire surface of the construction material.

[0226] Figure 4D depicts one embodiment of a first set of feed pulses comprising feed pulses 402-403 at an average feed rate of R6 (e.g.. 600 mL per minute) and a second set of feed pulses comprising feed pulses 404-405 at an average feed rate of R3 (e g., 300 mL per minute). The first set of feed pulses may be specified using a first feed rate profile or a first feed cycle profile. The second set of feed pulses may be specified using a second feed rate profile or a second feed cycle profile. The profiles may be stored in a memory, such as memory 171 in Figure IM.

[0227] Figure 4E depicts one embodiment of a first set of feed pulses 422 and a second set of feed pulses 423. As the time difference between the two sets of feed pulses corresponding to the time difference between times T3 and T9 is greater than a threshold period of time without moisture being added to a construction material or too long without liquid being applied to a construction material, a set of wetting pulses 424 comprising water may be applied to the construction material between times T6 and T7.

[0228] Figure 4F depicts one embodiment of a feed rate profile 452. Figure 4G depicts one embodiment of a feed rate profile 454.

[0229] Figure 4H depicts one embodiment of multiple feed cycles for applying a feed solution to a shaped structure. The multiple feed cycles include feed cycle 446 that includes seven feed pulses including feed pulse 447.

[0230] Figure 41 depicts one embodiment of multiple feed cycles for applying a feed solution to a shaped structure. The multiple feed cycles include feed cycle 442 that includes twelve pairs of feed pulses including feed pulse 443.

[0231] Figure 4J depicts a flowchart describing one embodiment of a process for manufacturing a construction material. In one embodiment, the process of Figure 4J is performed using one or more subsystems of the biostructure manufacturing system 100 depicted in Figure IB. In some embodiments, the process of Figure 4J is performed using a system for manufacturing a construction material that includes a climate controlled environment, such as the climate controlled environment 122 in Figures 1C-1E. and / or a feed system, such as the feed system 108 depicted in Figures 1A-1D, 10, and IP.

[0232] In step 462, mineralizing enzymes and / or mineralizing (e.g.. urease producing) microorganisms (e g., in an inoculation composition), and a plurality of aggregate particlesare acquired. In step 464, a shaped structure is generated or formed that includes the mineralizing (e.g., urease producing) microorganisms and the plurality of aggregate particles. The shaped structure may correspond with a brick or tile structure. In step 466, a first feed solution that includes a cementation reagent is acquired. In step 468, at least a threshold amount of a gas emanating from the shaped structure is detected. In step 470, the first feed solution is applied to the shaped structure using a first set of feed cycles with a first set of feed parameters. The application of the first feed solution to the shaped structure causes calcium carbonate to bind at least two particles of the plurality of aggregate particles. In step 472, a second feed solution that includes the cementation reagent is acquired. In step 474, the second feed solution is applied to the shaped structure using a second set of feed cycles with a second set of feed parameters. In step 476, a deodorizing process is applied to the shaped structure.

[0233] Figure 4K depicts a flowchart describing another embodiment of a process for manufacturing a construction material. In one embodiment, the process of Figure 4K is performed using one or more subsystems of the biostructure manufacturing system 100 depicted in Figure IB. In some embodiments, the process of Figure 4K is performed using a system for manufacturing a construction material that includes a climate controlled environment, such as the climate controlled environment 122 in Figures 1C-1E, and / or a feed system, such as the feed system 108 depicted in Figures 1A-1D, 10, and IP.

[0234] In step 482, mineralizing enzymes and / or mineralizing (e g., urease producing) microorganisms (e.g., in an inoculation composition), and a plurality of aggregate particles are acquired. In step 484, a shaped structure is acquired that includes the mineralizing (e.g., urease producing) microorganisms and the plurality of aggregate particles. In step 486, a feed solution that includes a cementation reagent is acquired. In step 488, an ambient temperature and an ambient humidity are determined. In step 490, a first set of feed parameters for a first set of feed cycles is determined based on the ambient temperature and the ambient humidity. In step 492, a pre-feed binder composition is applied to the shaped structure prior to application of the first feed solution to the shaped structure. In step 494, the first feed solution is applied to the shaped structure using the first set of feed cycles with the first set of feed parameters. The application of the first feed solution to the shaped structure causes calcium carbonate to bind at least two particles of the plurality of aggregate particles. In step 496. a finishing process is performed using the shaped structure.

[0235] Figure 5A depicts a flowchart describing another embodiment of a process for manufacturing a construction material. In one embodiment, the process of Figure 5A is performed using one or more subsystems of the biostructure manufacturing system 100 depicted in Figure IB. In some embodiments, the process of Figure 5 A is performed using a system for manufacturing a construction material that includes a climate controlled environment, such as the climate controlled environment 122 in Figures 1C-1E. and / or a feed system, such as the feed system 108 depicted in Figures I A- ID, 10, and IP.

[0236] In step 502, an aggregate mix is generated or acquired. The aggregate mix may be generated using a mixing tank to aggregate and mix a plurality of aggregate particles. In step 504, an inoculation (including mineralizing enzymes and / or microorganisms) composition is combined with the aggregate mix. In step 506, a portion of the aggregate mix is formed into a structure (e.g., into a shaped structure or into a tile structure). In step 508, a first feed solution with a first pH is acquired. In step 510. a first temperature and a first humidity for a first feed cycle are determined. In step 512, a fixation solution or a pre-feed solution is applied to the portion of the aggregate mix prior to applying the first feed solution. In step 514, the first feed solution is applied to the portion of the aggregate mix during the first feed cycle. In one embodiment, the first temperature and the first humidity for the first feed cycle may be determined based on a feed cycle number corresponding to the number of prior feed cycles that have been performed and / or the first pH of the first feed solution. In step 516, a second feed solution with a second pH greater than the first pH is acquired. In step 518. a second temperature and a second humidity for a second feed cycle are determined. In one embodiment, the second temperature and the second humidity for the second feed cycle may be determined based on a feed cycle number corresponding to the number of prior feed cycles that have been performed and / or the second pH of the second feed solution. In step 520. the second feed solution is applied to the portion of the aggregate mix during the second feed cycle. In step 522, a reinoculation cycle is performed subsequent to the second feed cycle.

[0237] Figure 5B depicts a flowchart describing another embodiment of a process for manufacturing a construction material. In one embodiment, the process of Figure 5B is performed using one or more subsystems of the biostructure manufacturing system 100 depicted in Figure IB. In some embodiments, the process of Figure 5B is performed using a system for manufacturing a construction material that includes a climate controlledenvironment, such as the climate controlled environment 122 in Figures 1C-1E, and / or a feed system, such as the feed system 108 depicted in Figures 1A-1D, 10, and IP.

[0238] In step 532, an aggregate mix is acquired. In step 534, an inoculation solution (including mineralizing enzymes and / or microorganisms) is applied to the aggregate mix. In step 536, a portion of the aggregate mix is formed into a shape (e.g., a structured shape). In step 538, a threshold level of ammonia gas is detected. In step 540, a crust formation process is applied to the portion of the aggregate mix in response to detection of the threshold level of ammonia gas. In step 542, an ambient temperature and an ambient humidity are determined. In step 544, a first feed solution with a first pH is identified based on the ambient temperature and ambient humidity. In step 546, the first feed solution is applied to the portion of the aggregate mix subsequent to the application of the crust formation process to the portion of the aggregate mix. In step 548, a residual pH for the first feed solution is determined. In step 550, a multi-pass feed process is applied to the portion of the aggregate mix based on the residual pH for the first feed solution.

[0239] Figure 5C depicts a flowchart describing another embodiment of a process for manufacturing a construction material. In one embodiment, the process of Figure 5C is performed using one or more subsystems of the biostructure manufacturing system 100 depicted in Figure IB. In some embodiments, the process of Figure 5C is performed using a system for manufacturing a construction material that includes a climate controlled environment, such as the climate controlled environment 122 in Figures 1C-1E, and / or a feed system, such as the feed system 108 depicted in Figures 1A-1D, 10, and IP.

[0240] In step 562, a plurality of aggregate particles is acquired. In step 564, the plurality of aggregate particles is formed into a shaped structure. In step 566, an ambient temperature and an ambient humidity’ are determined. In step 568, an average feed rate for a first feed cycle is determined based on the ambient temperature and the ambient humidity. In step 570, a maximum feed rate and a minimum feed rate for the first feed cycle are determined. In step 572, a first set of feed pulses for the first feed cycle is determined based on the average feed rate, the maximum feed rate, and the minimum feed rate. In step 574, the first set of feed pulses is applied to the shaped structure during the first feed cycle. In one example, the first set of feed pulses may correspond with feed pulses 435-437 in Figure 4C. In step 576, a second average feed rate for a second feed cycle is determined. In step 578, a second set of feed pulses for the second feed cycle is determined based on the secondaverage feed rate, the maximum feed rate, and the minimum feed rate. In step 580, the second set of feed pulses is applied to the shaped structure during the second feed cycle.

[0241] Figure 6A depicts a flowchart describing one embodiment of a process for manufacturing a construction material. In one embodiment, the process of Figure 6A is performed using one or more subsystems of the biostructure manufacturing system 100 depicted in Figure IB. In some embodiments, the process of Figure 6A is performed using a system for manufacturing a construction material that includes a climate controlled environment, such as the climate controlled environment 122 in Figures 1C-1E. and / or a feed system, such as the feed system 108 depicted in Figures 1A-1D, 10, and IP.

[0242] In step 602, a shaped structure that includes mineralizing enzymes and / or mineralizing (e.g., urease producing) microorganisms and a plurality of aggregate particles is provided. In step 604, a feed solution is acquired that has a molarity greater than a first molarity and a pH less than a first pH. The feed solution includes a cementation reagent. In step 606, the feed solution is applied to the shaped structure during one or more feed cycles, the application of the feed solution to the shaped structure causes calcium carbonate to bind at least two particles of the plurality of aggregate particles.

[0243] Figure 6B depicts a flowchart describing one embodiment of a process for manufacturing a construction material. In one embodiment, the process of Figure 6B is performed using one or more subsystems of the biostructure manufacturing system 100 depicted in Figure IB. In some embodiments, the process of Figure 6B is performed using a system for manufacturing a construction material that includes a climate controlled environment, such as the climate controlled environment 122 in Figures 1C-1E, and / or a feed system, such as the feed system 108 depicted in Figures 1A-1D, 10, and IP.

[0244] In step 612, mineralizing (e.g., urease producing) microorganisms and a plurality of aggregate particles are combined. In step 614, a shaped structure that includes the plurality of aggregate particles is formed or molded. In step 616, the shaped structure is hardened to a meta-stable state subsequent to or prior to combining the mineralizing (e.g., urease producing) microorganisms and the plurality of aggregate particles. In step 618, a feed solution that includes a cementation reagent is acquired. In step 620. subsequent to hardening of the shaped structure, the shaped structure is submersed in a bath of the feed solution. In step 622, the shaped structure is removed from the bath of the feed solution after at least 20% of the cementation reagent has been consumed.

[0245] In some embodiments, while a shaped structure is submersed in a liquid, the shaped structure is moved or vibrated to displace fluid that is present in the pores and replace with the fluid within the pores. The shaped structure may be submersed while both the shaped structure is moved within the fluid and the fluid is flowing at a particular flow rate. The fluid may be moved using pumps, movement of a container storing the fluid, or by causing waves in the fluid within the container (e.g., similar to generating waves in a wave pool). Waves caused by acoustic vibrations or mechanical means may push out unreacted material within the shaped structure. In some cases, the shaped structure may be moved along a conveyor belt while the shaped structure is fully immersed within the liquid. A technical benefit of displacing unreacted material is that a second bath may be avoided or multiple soaking phases followed by rinsing phases may be avoided.

[0246] In some embodiments, while a shaped structure is submersed in a liquid, a fluid flow of the liquid (e.g.. with a liquid velocity of at least 1 meter per minute) is applied to the construction material to displace fluid that is present in the pores and replace with the fluid within the pores.

[0247] Figure 6C depicts a flowchart describing one embodiment of a process for manufacturing a construction material. In one embodiment, the process of Figure 6C is performed using one or more subsystems of the biostructure manufacturing system 100 depicted in Figure IB. In some embodiments, the process of Figure 6C is performed using a system for manufacturing a construction material that includes a climate controlled environment, such as the climate controlled environment 122 in Figures 1C-1E, and / or a feed system, such as the feed system 108 depicted in Figures 1A-1D, 10, and IP.

[0248] In step 642, a mixture of mineralizing enzymes and / or mineralizing (e.g., urease producing) microorganisms (e.g., in an inoculation composition), and a plurality of aggregate particles is generated or acquired. In step 644, a shaped structure is formed using the mixture. In one example, the mixture is pressed into the shaped structure. The shaped structure may include a tile or brick structure. In step 646, the mixture is incubated subsequent to formation of the shaped structure. In step 648. a feed solution that includes a cementation reagent is acquired. In step 650, a topical suspension of a second inoculation composition is applied to the shaped structure prior to application of the feed solution to the shaped structure. In step 652, the feed solution is applied to the shaped structure during one or more feed cycles. The application of the feed solution to the shaped structure causes calcium carbonate to bind at least two particles of the plurality of aggregate particles. In step654, a deodorizing process is applied to the shaped structure subsequent to the application of the feed solution to the shaped structure.

[0249] In some cases, the spray / drip type systems are limited by the ability to flow fluid to the pores, which diminishes as the biocementation closes the pore throats. Adding a final soak step at the end, prior to the wash steps, could allow further cementation to occur as the mass transport mechanism is different: diffusion > fluid dynamics. We may use this to achieve enhanced structural strength for higher end applications.

[0250] Figure 23A depicts a flowchart describing another embodiment of a process for manufacturing a construction material. In one embodiment, the process of Figure 23A is performed using one or more subsystems of the biostructure manufacturing system 100 depicted in Figure IB or Figure 30A. In some embodiments, the process of Figure 23A is performed using a system for manufacturing a construction material that includes a climate controlled environment, such as the climate controlled environment 122 in Figures 1C-1E, and / or a feed system, such as the feed system 108 depicted in Figures 1 A-1D. 10, and IP.

[0251] In step 2302 a shaped structure including a plurality of aggregate particles and one or more mineralizing enzymes and / or microorganisms and having a pore volume is provided. In step 2304, a feed solution that includes a cementation reagent is provided, a volume of which is applied solution to the shaped structure in step 2306 that is from 0.5x to 1.5x the pore volume. In step 2308, the shaped structure is cured for a time sufficient to cause or allow a portion of a cementation reagent in the feed solution to be converted to mineral binder in the pore volume. In step 2310, steps 2306 and 2308 are repeated.

[0252] Figure 23B depicts a flowchart describing another embodiment of a process for manufacturing a construction material. In one embodiment, the process of Figure 23B is performed using one or more subsystems of the biostructure manufacturing system 100 depicted in Figure IB or Figure 30A. In some embodiments, the process of Figure 23B is performed using a system for manufacturing a construction material that includes a climate controlled environment, such as the climate controlled environment 122 in Figures 1C-1E, and / or a feed system, such as the feed system 108 depicted in Figures 1 A-1D, 10, and IP

[0253] In step 2322 a shaped structure including a plurality of aggregate particles and one or more mineralizing enzymes and / or microorganisms and having a pore volume is provided. In step 2324, a feed solution that includes a cementation reagent is provided, a volume of which is applied solution to the shaped structure in step 2326 in one or moresingle-pass feed cycles. In step 2328, the shaped structure is cured for a time sufficient to cause or allow a portion of a cementation reagent in the feed solution to be converted to mineral binder in the pore volume. Steps 2326 and 2328 may be repeated.

[0254] Figure 23C depicts a flowchart describing another embodiment of a process for manufacturing a construction material. In one embodiment, the process of Figure 23C is performed using one or more subsystems of the biostructure manufacturing system 100 depicted in Figure IB or Figure 30A. In some embodiments, the process of Figure 23C is performed using a system for manufacturing a construction material that includes a climate controlled environment, such as the climate controlled environment 122 in Figures 1C-1E, and / or a feed system, such as the feed system 108 depicted in Figures 1 A-1D, 10, and IP.

[0255] In step 2332 a shaped structure including a plurality of aggregate particles and one or more mineralizing enzymes and / or microorganisms and having a pore volume is provided. In step 2334, a feed solution that includes a cementation reagent is provided, a volume of which is applied solution to the shaped structure in step 2336 that is from 0.5x to 1.5x the pore volume. In step 2338, the shaped structure is cured for a time sufficient to cause or allow a portion of a cementation reagent in the feed solution to be converted to mineral binder in the pore volume. In step 2340 an nth volume of the feed solution to the shaped structure that is from 0.5x to l.Sx the nth pore volume is applied. In step 2310, steps 2306 and 2308 are repeated.

[0256] Figure 24A depicts a flowchart describing another embodiment of a process for manufacturing a construction material. In one embodiment, the process of Figure 24A is performed using one or more subsystems of the biostructure manufacturing system 100 depicted in Figures 1A-1D, 10, IP, and 31A-31I. In some embodiments, the process of Figure 24A is performed using a system for manufacturing a construction material that includes a feed system, such as the feed system 108 depicted in Figures 1A-1D, 10, IP, and 31A-31I.

[0257] In step 2422 a shaped structure including a plurality of aggregate particles and one or more mineralizing enzymes and / or microorganisms and having a pore volume is provided. In step 2424, a feed solution that includes a cementation reagent of a high molarity (e.g.. multiple high molarity cementation reagents) is provided, which is applied to the aggregate particles in step 2426. In step 2428, the plurality of aggregate particles is curedso that a mineral binder is formed by action of the mineralizing enzymes and / or microorganisms consolidates the plurality of aggregate particles.

[0258] Figure 24B depicts a flowchart describing another embodiment of a process for manufacturing a construction material. In one embodiment, the process of Figure 24B is performed using one or more subsystems of the biostructure manufacturing system 100 depicted in Figures 1A-1D, 10, IP, and 31A-31I. In some embodiments, the process of Figure 24B is performed using a system for manufacturing a construction material that includes a feed system, such as the feed system 108 depicted in Figures 1A-1D, 10, IP, and 31A-31I.

[0259] In step 2432 a plurality of aggregate particles (e.g., in an aggregate blend) mixed with one or more mineralizing enzymes and / or microorganisms. In step 2434, a feed solution including a cementation reagent and having an osmolarity of at least 4.1 Osm / L is provided, which is applied to the aggregate particles in step 2436. In step 2438, the plurality of aggregate particles is cured so that a mineral binder is formed by action of the mineralizing enzymes and / or microorganisms consolidates the plurality of aggregate particles.

[0260] Figure 24C depicts a flowchart describing another embodiment of a process for manufacturing a construction material. In one embodiment, the process of Figure 24C is performed using one or more subsystems of the biostructure manufacturing system 100 depicted in Figures 1A-1D, 10, IP, and 31A-31I. In some embodiments, the process of Figure 24C is performed using a system for manufacturing a construction material that includes a feed system, such as the feed system 108 depicted in Figures 1A-1D, 10, IP, and 31A-31I.

[0261] In step 2442 a plurality of aggregate particles (e.g., in an aggregate blend) mixed with one or more mineralizing enzy mes and / or microorganisms. In step 2444, a feed solution including a cementation reagent and having an osmotic pressure of at least 15 MPa and / or an ionic strength of at least 2 (e.g., at least 4.6, e.g., at least 5) moles per liter, which is applied to the aggregate particles in step 2446. In step 2448, the plurality of aggregate particles is cured so that a mineral binder is formed by action of the mineralizing enzymes and / or microorganisms consolidates the plurality of aggregate particles.

[0262] Figure 24D depicts a flowchart describing another embodiment of a process for manufacturing a construction material. In one embodiment, the process of Figure 24D isperformed using one or more subsystems of the biostructure manufacturing system 100 depicted in Figures 1A-1D, 10, IP, and 31A-31I. In some embodiments, the process of Figure 24D is performed using a system for manufacturing a construction material that includes a feed system, such as the feed system 108 depicted in Figures 1A-1D, 10, IP, and 31A-31I.

[0263] In step 2452 a plurality of aggregate particles (e.g., in an aggregate blend) mixed with one or more mineralizing enzy mes and / or microorganisms. In step 2454, a feed solution including a cementation reagent and having a pH of between 6 and 2, which is applied to the aggregate particles in step 2456. In step 2458, the plurality of aggregate particles is cured so that a mineral binder is formed by action of the mineralizing enzymes and / or microorganisms consolidates the plurality of aggregate particles.

[0264] Figure 25A depicts a flowchart describing another embodiment of a process for manufacturing a construction material. In one embodiment, the process of Figure 25A is performed using one or more subsystems of the biostructure manufacturing system 100 depicted in Figures 1A-1D, 10, IP, and 31A-31I. In some embodiments, the process of Figure 25A is performed using a system for manufacturing a construction material that includes a feed system, such as the feed system 108 depicted in Figures 1A-1D, 10, IP, and 31A-31I.

[0265] In step 2502 a mixture of aggregate particles and a population of mineralizing microorganisms is generated. In step 2504, the mixture is incubated under conditions that expand the population of mineralizing microorganisms. In step 2506, a cementation reagent solution to the shaped structure.

[0266] Figure 25B depicts a flowchart describing another embodiment of a process for manufacturing a construction material. In one embodiment, the process of Figure 25B is performed using one or more subsystems of the biostructure manufacturing system 100 depicted in Figures 1A-1D, 10, IP. and 31A-31I. In some embodiments, the process of Figure 25B is performed using a system for manufacturing a construction material that includes a feed system, such as the incubation system 106 of Figures 1A-1D, 10, IP and / or the crust formation system of Figure ID, and Figure 30A, and the feed system 108 depicted in Figures 1A-1D, 10, IP, and 31A-31I.

[0267] In step 2512 a mixture of aggregate particles and a population of mineralizing microorganisms is generated. In step 2514, the mixture is formed into a shaped structure. Instep 2516, the shaped structure is incubated at a temperature and / or air-flow that both expands a population of the mineralizing microorganisms and forms a stabilizing crust.

[0268] Figure 25C depicts a flowchart describing another embodiment of a process for manufacturing a construction material. In one embodiment, the process of Figure 25C is performed using one or more subsystems of the biostructure manufacturing system 100 depicted in Figures 1A-1D, 10, IP, and 31A-31I. In some embodiments, the process of Figure 25C is performed using a system for manufacturing a construction material that includes a feed system, such as the incubation system 106 of Figures 1A-1D, 10, IP and / or the crust formation system of Figure ID, and Figure 30A, and the feed system 108 depicted in Figures 1A-1D, 10, IP, and 31A-31I.

[0269] In step 2522 a mixture of aggregate particles and a population of mineralizing microorganisms is generated. In step 2524, the mixture is formed into a shaped structure. In step 2526. the shaped structure is incubated under conditions that both expand a population of the mineralizing microorganisms and forms a stabilizing crust. In step 2528 a cementation reagent solution is applied to the shaped structure.

[0270] Figure 25D depicts a flowchart describing another embodiment of a process for manufacturing a construction material. In one embodiment, the process of Figure 25D is performed using one or more subsystems of the biostructure manufacturing system 100 depicted in Figures 1A-1D, 10, IP, and 31A-31I. In some embodiments, the process of Figure 25D is performed using a system for manufacturing a construction material that includes a feed system, such as the incubation system 106 of Figures 1A-1D, 10, IP and / or the crust formation system of Figure ID, and Figure 30A, and the feed system 108 depicted in Figures 1A-1D, 10, IP, and 31A-31I.

[0271] In step 2532 a mixture of aggregate particles and a population of mineralizing microorganisms is generated. In step 2534, the mixture is formed into a shaped structure. In step 2536. for a time sufficient to expand a population of the mineralizing microorganisms. In step 2538 a cementation reagent solution is applied to the shaped structure.

[0272] Figure 25E depicts a flowchart describing another embodiment of a process for manufacturing a construction material. In one embodiment, the process of Figure 25E is performed using one or more subsystems of the biostructure manufacturing system 100 depicted in Figures 1A-1D, 10, IP, 31A-31I, 32A-32I. In some embodiments, the process of Figure 25E is performed using a system for manufacturing a construction material thatincludes a biologies activation and / or expansion system, such as depicted in Figure 30B and 30C, a feed system, such as the incubation system 106 of Figures 1 A-1D, 10, IP and / or the crust formation system of Figure ID, and Figure 30A, and the feed system 108 depicted in Figures 1A-1D, 10, IP, and 31A-31I.

[0273] In step 2542 a population of mineralizing microorganisms to form an inoculation solution. In step 2544, an aggregate blend is inoculated with the activated population of mineralizing microorganisms to make an inoculated aggregate blend. In step 2546, the inoculated aggregate blend is formed into one or more shaped stmcture(s). In step 2548, the population of mineralizing microorganisms is caused or allowed to expand in the one or more shaped structures. In step 2550 a cementation reagent solution is applied to the shaped structure to form a mineral binder in the aggregate blend.

[0274] Figure 25F depicts a flowchart describing another embodiment of a process for manufacturing a construction material. In one embodiment, the process of Figure 25F is performed using one or more subsystems of the biostructure manufacturing system 100 depicted in Figures 1A-1D, 10, IP, 31A-31I, 32A-32I. In some embodiments, the process of Figure 25F is performed using a system for manufacturing a construction material that includes a biologies activation and / or expansion system, such as depicted in Figure 30B and 30C, a feed system, such as the incubation system 106 of Figures 1A-1D, 10, IP and / or the crust formation system of Figure ID. and Figure 30A. and the feed system 108 depicted in Figures 1A-1D, 10, IP, and 31A-31I.

[0275] In step 2572 a population of mineralizing microorganisms to form an inoculation solution. In step 2574, an aggregate blend is inoculated with the activated population of mineralizing microorganisms to make an inoculated aggregate blend. In step 2576, the population of mineralizing microorganisms is caused or allowed to expand in the one or more shaped structures. In step 2578, the inoculated aggregate blend is formed into one or more shaped structure(s). In step 2580, the population of mineralizing microorganisms is caused or allowed to expand in the one or more shaped structures. In step 2582 a cementation reagent solution is applied to the shaped structure to form a mineral binder in the aggregate blend.

[0276] Figure 25G depicts a flowchart describing another embodiment of a process for manufacturing a construction material. In one embodiment, the process of Figure 25G is performed using one or more subsystems of the biostructure manufacturing system 100depicted in Figures 1A-1D, 10, IP, 31A-31I, 32A-32I. In some embodiments, the process of Figure 25G is performed using a system for manufacturing a construction material that includes a biologies activation and / or expansion system, such as depicted in Figure 30B and 30C, a feed system, such as the incubation system 106 of Figures 1 A-1D, 10, IP and / or the crust formation system of Figure ID, and Figure 30A, and the feed system 108 depicted in Figures 1A-1D, 10. IP. and 31A-31I.

[0277] In step 2592 a population of mineralizing microorganisms to form seed solution. In step 2594, the seed solution is added to a fermentation solution and expand the population of mineralizing microorganisms to form an inoculation solution. In step 2596, an aggregate blend is inoculated with the inoculation solution. In step 2598, the inoculated aggregate blend is formed into one or more shaped structure(s). In step 25100, the population of mineralizing microorganisms is caused or allowed to expand in the one or more shaped structures. In step 25102. a cementation reagent solution is applied to the shaped structure to form a mineral binder in the aggregate blend.

[0278] Figure 25H depicts a flowchart describing another embodiment of a process for manufacturing a construction material. In one embodiment, the process of Figure 25H is performed using one or more subsystems of the biostructure manufacturing system 100 depicted in Figures 1A-1D, 10, IP, 31A-31I, 32A-32I. In some embodiments, the process of Figure 25H is performed using a system for manufacturing a construction material that includes a biologies activation and / or expansion system, such as depicted in Figure 30B and 30C, a feed system, such as the incubation system 106 of Figures 1 A-1D, 10, IP and / or the crust formation system of Figure ID, and Figure 30A, and the feed system 108 depicted in Figures 1 A- ID. 10. IP. and 31A-31I.

[0279] In step 25112 a population of mineralizing microorganisms to form seed solution. In step 25114, the seed solution is added to a fermentation solution and expand the population of mineralizing microorganisms to form an inoculation solution. In step 25116, an aggregate blend is inoculated with the inoculation solution. In step 25118, the population of mineralizing microorganisms is caused or allowed to expand in the one or more shaped structures. In step 25120, the inoculated aggregate blend is formed into one or more shaped structure(s). In step 25122, the population of mineralizing microorganisms is caused or allowed to expand in the one or more shaped structures. In step 25124, a cementation reagent solution is applied to the shaped structure to form a mineral binder in the aggregate blend.

[0280] Figure 26A depicts a flowchart describing another embodiment of a process for manufacturing a construction material. In one embodiment, the process of Figure 26A is performed using one or more subsystems of the biostructure manufacturing system 100 depicted in Figures 1A-1D, 10, IP, and 31A-31I.

[0281] In step 2602 an aggregate blend having an unpressed green pycnometric intergranular void percentage or porosity of less than 55% (e.g., less than 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, or 10%) is provided. In step 2604, the aggregate blend is mixed with a mineralizing enzyme or mineralizing microorganism. In step 2606, a cementation reagent is provided to form a mineral binder.

[0282] Figure 26B depicts a flowchart describing another embodiment of a process for manufacturing a construction material. In one embodiment, the process of Figure 26B is performed using one or more subsystems of the biostructure manufacturing system 100 depicted in Figures 1A-1D, 10. IP, and 31A-31I.

[0283] In step 2612 an aggregate blend of two or more aggregates in proportions that correspond within 10% to a maximum theoretical packing density for the aggregate blend is provided. In step 2614, the aggregate blend is mixed with a mineralizing enzyme or mineralizing microorganism. In step 2616, a cementation reagent is provided to form a mineral binder.

[0284] Figure 26C depicts a flowchart describing another embodiment of a process for manufacturing a construction material. In one embodiment, the process of Figure 26C is performed using one or more subsystems of the biostructure manufacturing system 100 depicted in Figures 1A-1D. 10. IP. and 31A-311.

[0285] In step 2622 an aggregate blend is provided of two or more aggregates having a maximum particle size, D, a gradation curve on a plot of % of particles passing through a sieve of a sieve size vs the sieve size of pi = (di / D)045(Equation A) or p, = (di I D)05(Equation B), or within a least squares error of 0.00001 to 0.1 of the gradation curve of Equation A or the gradation curve of Equation B, where pi is a % of particles passing through an ith sieve size, and di is the opening size of the ith sieve size. In step 2624. the aggregate blend is mixed with a mineralizing enzyme or mineralizing microorganism. In step 2626, a cementation reagent is provided to form a mineral binder.

[0286] Figure 26D depicts a flowchart describing another embodiment of a process for manufacturing a construction material. In one embodiment, the process of Figure 26D isperformed using one or more subsystems of the biostructure manufacturing system 100 depicted in Figures 1A-1D, 10. IP, and 31A-31I.

[0287] In step 2632 an aggregate blend of two or more aggregates that is a gap-graded aggregate blend is provided. In step 2634, the aggregate blend is mixed with a mineralizing enzy me or mineralizing microorganism. In step 2636, a cementation reagent is provided to form a mineral binder.

[0288] Figure 26E depicts a flowchart describing another embodiment of a process for manufacturing a construction material. In one embodiment, the process of Figure 26E is performed using one or more subsystems of the biostructure manufacturing system 100 depicted in Figures 1A-1D. 10. IP, and 31A-31I.

[0289] In step 2642 an aggregate blend is provided of two or more aggregates having a maximum particle size, D, a gradation curve on a plot of % of particles passing through a sieve of a sieve size vs the sieve size, and having an area thereunder that is ±20% of an area under a gradation curve of p, = (di I D)045(Equation A) or pi = (di / D)°5(Equation B), where pi is a % of particles passing through an ith sieve size, and di is the opening size of the ith sieve size. In step 2644, the aggregate blend is mixed with a mineralizing enzyme or mineralizing microorganism. In step 2646, a cementation reagent is provided to form a mineral binder.

[0290] Figure 26F depicts a flowchart describing another embodiment of a process for manufacturing a construction material. In one embodiment, the process of Figure 26F is performed using one or more subsystems of the biostructure manufacturing system 100 depicted in Figures 1A-1D. 10. IP. and 31A-311.

[0291] In step 2652 an aggregate blend of two or more aggregates having a water infiltration rate (ksat) of between 2 and 500 ml / min is provided. In step 2654, the aggregate blend is mixed with a mineralizing enzyme or mineralizing microorganism. In step 2656, a cementation reagent is provided to form a mineral binder.

[0292] Figure 27A depicts a flowchart describing another embodiment of a process for manufacturing a construction material. In one embodiment, the process of Figure 27A is performed using one or more subsystems of the biostructure manufacturing system 100 depicted in Figures 1A-1D, 10, IP, and 31A-31I.

[0293] In step 2702 a plurality' of aggregate particles (e.g., an aggregate blend) including 0.1-2 wt% of organic or inorganic fibers is provided. In step 2704, the plurality of aggregateparticles is mixed with a mineralizing enzyme or mineralizing microorganism. In step 2706, a cementation reagent is provided to form a mineral binder.

[0294] Figure 27B depicts a flowchart describing another embodiment of a process for manufacturing a construction material. In one embodiment, the process of Figure 27B is performed using one or more subsystems of the biostructure manufacturing system 100 depicted in Figures 1A-1D, 10, IP, and 31A-31I.

[0295] In step 2712 an aggregate blend is provided. In step 2714, the aggregate blend is mixed with a mineralizing enzyme or mineralizing microorganism. In step 2716, the mixture is formed into a shaped structure around one or more strain-enhancing textiles. In step 2718 a cementation reagent is provided to form a mineral binder. In some embodiments, the strain-enhancing textile(s) are disposed within a tension zone of the construction material.

[0296] Figure 27C depicts a flowchart describing another embodiment of a process for manufacturing a construction material. In one embodiment, the process of Figure 27C is performed using one or more subsystems of the biostructure manufacturing system 100 depicted in Figures 1A-1D, 10, IP, and 31A-31I.

[0297] In step 2722 one or more aggregates is combined with calcium carbonate particles and a metal hydroxide such that a portion of the calcium carbonate particles are fused by reaction with the metal hydroxide to produce fused metal carbonates. In step 2724, the one or more aggregates is mixed with a mineralizing enzyme or mineralizing microorganism. In step 2726, the mixture is formed into a shaped structure around one or more strain-enhancing textiles. In step 2718 a cementation reagent is provided to form a mineral binder.

[0298] Figure 27D depicts a flowchart describing another embodiment of a process for manufacturing a construction material. In one embodiment, the process of Figure 27D is performed using one or more subsystems of the biostructure manufacturing system 100 depicted in Figures 1A-1D, 10, IP, and 31A-31I.

[0299] In step 2732 a combination of one or more aggregates with magnesium hydroxide (or a source of magnesium hydroxide) and one or more mineralizing enzyme(s) and / or mineralizing microorganism(s) is provided. In step 2734. one or more sources of carbonate ions to the combination to produce dissolved carbonate ions is applied. In step 2736, dissolved metal ions are applied to and / or generated in the combination. In step 2738 the dissolved carbonate ions are caused or allowed to react with the magnesium hydroxideand the metal ions in the combination to form a mineral binder that consolidates the combination.

[0300] Figure 27E depicts a flowchart describing another embodiment of a process for manufacturing a construction material. In one embodiment, the process of Figure 27E is performed using one or more subsystems of the biostructure manufacturing system 100 depicted in Figures 1A-1D, 10, IP, and 31A-31I.

[0301] In step 2742 a combination of one or more aggregates with magnesium hydroxide and one or more mineralizing enzyme(s) and / or mineralizing microorganism(s). In step 2744, a source of carbonate ions to the combination to produce dissolved carbonate ions by action of the one or more mineralizing enzyme(s) and / or mineralizing microorganism(s) is applied. In step 2746, dissolved metal ions are applied to and / or generated in the combination. In step 2748 gaseous carbon dioxide to the combination is applied to form dissolved carbonate ions in the combination. In step 2750 the dissolved carbonate ions to react with the magnesium hydroxide and the metal ions in the combination to form a mineral binder that consolidates the combination. In some embodiments, the magnesium hydroxide is formed by reaction of magnesium sulfate in the aggregate with hydroxide ions dissolved from caustic components of the one or more aggregates.

[0302] Figure 27F depicts a flowchart describing another embodiment of a process for manufacturing a construction material. In one embodiment, the process of Figure 27F is performed using one or more subsystems of the biostructure manufacturing system 100 depicted in Figures 1A-1D, 10, IP, and 31A-31I.

[0303] In step 2752 a plurality of aggregate particles is provided. In step 2754, the plurality of aggregate particles is mixed with a mineralizing enzyme or mineralizing microorganism. In step 2756, the plurality of aggregate particles is combined with one or more additive(s) that: resists ice crystal formation; and / or induces micropore formation and / or resists water penetration. In step 2758, a cementation reagent is provided to form a mineral binder. In some embodiments, step 2758 is performed prior to step 2756. In some embodiments, step 2756 is performed after forming the plurality aggregate particles into a shape. In some embodiments, step 2756 is performed after curing. In some embodiments, step 2756 is performed before or after honing. In some embodiments, step 2756 is performed before or after polishing.

[0304] Figure 28A depicts a flowchart describing another embodiment of a process for manufacturing a construction material. Figure 28A depicts a flowchart describing another embodiment of a process for manufacturing a concentrated ammonium composition. In one embodiment, the process of Figure 28A is performed using one or more subsystems of the regeneration system 3009 depicted in Figures 31A, 31D, or 32A-32I and / or an effluent system such as depicted in Figures 31 A, 31C, or 3 ID.

[0305] In step 2802 a dilute aqueous solution of an ammonium salt and one or more organic compounds produced by a mineralizing microorganism is provided. In step 2804, the dilute solution is concentrated by reverse osmosis; forward osmosis; evaporation; precipitation by adding a higher solubility salt of a same counterion to the ammonium salt; or a combination of these. In some embodiments, the ammonium salt is concentrated at least partially in a regeneration system 3009.

[0306] Figure 28B depicts a flowchart describing another embodiment of a process for manufacturing a construction material. Figure 28B depicts a flowchart describing another embodiment of a process for manufacturing a concentrated ammonium composition. In one embodiment, the process of Figure 2BA is performed using one or more subsystems of the regeneration system 3009 depicted in Figures 31A, 31D, or 32A-32I and / or an effluent system such as depicted in Figures 31 A, 31C, or 3 ID.

[0307] In step 2812 an aqueous solution of ammonium chloride and one or more organic compounds produced by a mineralizing microorganism is provided. In step 2814, concentrated sulfuric acid is added to the solution to generate ammonium sulfate and evolve hydrogen chloride. In step 2816 the hydrogen chloride is collected and / or combined with calcium carbonate to produce calcium chloride and carbon dioxide. In step 2818, the dilute ammonium sulfate solution is concentrated by reverse osmosis; forward osmosis; evaporation; precipitation by adding a higher solubility salt of a same counterion to the ammonium salt; or a combination of these. In some embodiments, the ammonium chloride is concentrated at least partially in a regeneration system 3009.

[0308] Figures 29A-29F depict flowcharts describing embodiments of a process for manufacturing a construction material. In embodiments, the processes of Figures 29A-29F may be performed using one or more subsystems of the biostructure manufacturing system 100 depicted in Figures 1A-1D, 10, IP, 31A-31I, 32A-32I. In some embodiments, the processes of Figures 29A-29F may be performed using a system for manufacturing aconstruction material that includes a biologies activation and / or expansion system, such as depicted in Figure 30B and 30C, a feed system, such as the incubation system 106 of Figures 1A-1D, 10, IP and / or the crust formation system of Figure ID, and Figure 30A, and the feed system 108 depicted in Figures 1A-1D, 10, IP, and 31A-31I.

[0309] In step 2902 a population of mineralizing cells and / or cell spores is provided. In step 2904, the population of cell spores is germinated and / or the population of cells activated and / or expanded in an environment that induces a stress response (e.g., a high osmolality liquid medium, a temperature stress environment, a nutrient-deficient medium, or an oxidative stress environment, a low moisture environment, etc.) such that a resulting population of cells is metabolically primed to produce one or more osmoprotectants.

[0310] Figure 29B depicts a flowchart describing another embodiment of a process for manufacturing a construction material. In step 2912 a population of mineralizing cells and / or cell spores is provided. In step 2914, the population of cell spores is germinated and / or the population of cells activated and / or expanded in a high osmolality liquid medium such that a resulting population of cells is metabolically primed to produce one or more osmoprotectants to tolerate a high osmolality feeding solution including a source of calcium ions. In step 2916, the metabolically primed cells are combined with aggregate particles to form a mixture. In step 2918 a cementation reagent is provided to the mixture form a mineral binder by action of the metabolically primed mineralizing cells on the cementation reagent.

[0311] Figure 29C depicts a flowchart describing another embodiment of a process for manufacturing a construction material. In step 2922 a population of mineralizing cells and / or cell spores is provided. In step 2924, population of mineralizing cells and / or cell spores is combined with aggregate particles and a high osmolality liquid medium to form a mixture. In step 2926. the population of cells is expanded (e.g., proliferates) in the aggregate particles with the high osmolality liquid medium such that a resulting expanded / proliferated population of cells is metabolically primed to produce or uptake one or more osmoprotectants to tolerate a high osmolality feeding solution including a cementation reagent (e.g., a source of calcium ions, e.g., calcium chloride). In step 2928 a cementation reagent is provided to the mixture form a mineral binder by action of the metabolically primed mineralizing cells on the cementation reagent.

[0312] Figure 29D depicts a flowchart describing another embodiment of a process for manufacturing a construction material. In step 2932 an aggregate blend including apopulation of mineralizing cells and / or cell spores is provided. In step 2934, a feed solution is provided including a cementation reagent and one or more osmoprotectants. In step 2936, a mineral binder is formed in the aggregate by action of mineralizing cells on the cementation reagent. Alternatively, or in addition, the one or more osmoprotectants may be provided with the population of mineralizing cells (e.g., as part of a dried biologies formulation), and / or mixed in the aggregate blend.

[0313] Figure 29E depicts a flowchart describing another embodiment of a process for manufacturing a construction material. In step 2942 a population of mineralizing cells and / or cell spores is provided. In step 2944, the population of cell spores is germinated and / or the population of cells is expanded in a germination / fermentation medium including one or more osmoprotectants. In step 2946, the population of germinated / expanded cells is combined with aggregate particles to form a mixture. In step 2948. a cementation reagent to the mixture form a mineral binder by action of the mineralizing cells on the cementation reagent.

[0314] Figure 29F depicts a flowchart describing another embodiment of a process for manufacturing a construction material. In step 2952 a population of mineralizing cells and / or cell spores is provided. In step 2954, the population of mineralizing cells and / or cell spores is combined with an aggregate blend and one or more osmoprotectant to form a mixture. In step 2956, the population of cells in the mixture is expanded. In step 2958. a cementation reagent to the mixture form a mineral binder by action of the mineralizing cells on the cementation reagent.

[0315] Figure 30A and 30D depict embodiments of a biomanufacturing system 100. The biomanufacturing system 100 may include one or more of: a biologies activation system 3001, a mixing tank 102 (e.g., a cement mixing tank), a staging tank 3005 (e.g.. coupled to a system of pumps), an incubation system 106 (e.g., including one or more heat sources, sensors, humidifiers, fans, filters, etc.), a computing system 101, one or more pumps 3012, a conveyor system 120 (e.g., including a conveyor belt, a carousel, robotic arm conveyor, a monorail conveyor, an overhead conveyor, and / or a crane), a first cementation reagent source (e.g., a first cementation reagent tank), a second cementation reagent source, an acid source (e.g., an acid tank, e.g.. configured to store hydrochloric or sulfuric acid, e.g., coupled to a system of pipes and pumps, e.g., connecting to the staging tank), a pH sensor 124. a concentration sensor 125 (e.g., an EC sensor), a temperature sensor 109 (e.g., athermocouple, a thermistor, or a semiconductor-based sensor), a humidity sensor, a heating and / or cooling system 3007 (e.g., including one or more fans, heating elements, cooling elements, etc.), a gas source (e.g., an oxygen source, a carbon dioxide source, etc., e.g., fluidly coupled to the biologies activation and / or expansion tank and / or the feed system), a clean-in-place system 3010 (e.g., including a tank configured to store one or more CIP reagents, a system of pumps and pipes to connect to. e.g., the staging tank, the feed system, an effluent system, etc.), an effluent system 3008 (e.g., including one or more filters (e.g., microfilters), a biologies inactivation system, an effluent tank, an effluent drain valve, etc.), and / or a regeneration system (3009) (e.g.. including one or more permeable barriers, pumps, mixers, etc.). The biomanufacturing system 100 may also include one or more of: a spray system; a tile curing system; an optical sensor; a light source; or a feeding control application 192 in the computing system 101, for example, to control a feed rate based on observed infiltration rate. The biomanufacturing system may also include a forming system 104 (e.g. , a press (e.g., a vibratory press) and / or mold), a finishing system 114 / 3006 (e.g., a rinsing system (e.g., including a water source and one or more spray nozzles, baths, etc.), a drying system (e.g., including one or more fans), ahoning device, apolishing device, etc.), a pre-feed crust formation application 191 in the computing system, an environmental control application 194 in the computing system 101, or a conveyor control application 193 in the computing system 101. The biomanufacturing system 100 may further include one or more of: a feed bath 3101, an impulse delivery system 3103 (e.g., including a piston, an actuator, a plow, a diaphragm, a screw, or a blade, and / or a mechanism for positionally translating the shaped structure, e.g., one or more components of the conveyor system (e.g.. the in-bath conveyor system), e.g., a conveyor belt, carouse, or crane), an in-bath conveyor system 3102, an inter-system conveyor system (3101) (e.g., including a finger car, a conveyor belt, and / or a robotic arm), an impulse control application (195) in the computing system (101); or a regeneration control application (196) in the computing system 101. Black arrows show primarily solid mass flows (e.g., aggregate or shaped structures) progressing through the system. White arrows show primarily liquid mass flows (e.g., reagent solutions, feed solution, acid, effluent, etc.) progressing through the system. The dashed arrow shows a return of liquid to the feed system in the event that the regeneration system is used.

[0316] Figures 30B and 30C depict embodiments of a biologies activation and / or expansion system. These systems may include activation and / or expansion tank 3015, a heating / cooling system 3007, and optionally comprising one or more sensors, one or more transmitters, a sample port 3017, an enzymatic activity monitoring system 3018, mixer(s) 3016, pump(s) 3012.

[0317] Figures 31A-31I depict embodiments of a biomanufacturing system for performing a soak-based feeding process. The biomanufacturing system 100 may include one or more of: a biologies activation system 3001, a mixing tank 102 (e.g.. a cement mixing tank), a staging tank 3005 (e.g., coupled to a system of pumps), an incubation system 106 (e.g., including one or more heat sources, sensors, humidifiers, fans, filters, etc.), a computing system 101, one or more pumps 3012, a conveyor system 120 (e.g., including a conveyor belt, a carousel, robotic arm conveyor, a monorail conveyor, an overhead conveyor, and / or a crane), a first cementation reagent source (e.g.. a first cementation reagent tank), a second cementation reagent source, an acid source (e.g., an acid tank, e.g., configured to store hydrochloric or sulfuric acid, e.g., coupled to a system of pipes and pumps, e.g., connecting to the staging tank), a pH sensor 124, a concentration sensor 125 (e.g., an EC sensor), a temperature sensor 109 (e.g.. a thermocouple, a thermistor, or a semiconductor-based sensor), ahumidity sensor, aheating and / or cooling system 3007 (e.g., including one or more fans, heating elements, cooling elements, etc.), a gas source (e.g., an oxygen source, a carbon dioxide source, etc., e.g., fluidly coupled to the biologies activation and / or expansion tank and / or the feed system), a clean-in-place system 3010 (e.g., including a tank configured to store one or more CIP reagents, a system of pumps and pipes to connect to, e.g., the staging tank, the feed system, an effluent system, etc.), an effluent system 3008 (e.g., including one or more filters (e.g., microfilters), a biologies inactivation system, an effluent tank, an effluent drain valve, etc.), and / or a regeneration system (3009) (e.g., including one or more permeable barriers, pumps, mixers, etc.). The biomanufacturing system may also include a forming system 104 (e.g. , a press (e.g., a vibratory press) and / or mold), a finishing system 114 / 3006 (e.g., a rinsing system (e.g., including a water source and one or more spray nozzles, baths, etc.), a drying system (e.g., including one or more fans), a honing device, a polishing device, etc.), a pre-feed crust formation application 191 in the computing system, an environmental control application 194 in the computing system 101, or a conveyor control application 193 in the computing system 101. The biomanufacturing system 100 may further include one or more of: a feed bath 3101, animpulse delivery system 3103 (e.g., including a piston, an actuator, a plow, a diaphragm, a screw, or a blade, and / or a mechanism for positionally translating the shaped structure, e.g., one or more components of the conveyor system (e.g., the in-bath conveyor system), e.g., a conveyor belt, carouse, or crane), an in-bath conveyor system 3102, an inter-system conveyor system (3101) (e.g., including a finger car, a conveyor belt, and / or a robotic arm), an impulse control application (195) in the computing system (101); or a regeneration control application (196) in the computing system 101. Black arrows show primarily solid mass flows (e.g., aggregate or shaped structures) progressing through the system. White arrows show primarily liquid mass flows (e.g., reagent solutions, feed solution, acid, effluent, etc.) progressing through the system. Black and white striped arrows show mixed liquid-solid mass flows, or mass flows that could be liquid or solid. The feeding system may be configured for convective mixing, e.g., by having a high density reagent added to a top portion of the feed bath, and a low density reagent added to the bottom of the feed bath, e.g., as shown in Figure 311.

[0318] Figures 31J-31P depict flowcharts describing embodiments of a process for manufacturing a construction material. In embodiments, the processes of Figures 29A-29F may be performed using one or more subsystems of the biostructure manufacturing system 100 depicted in Figures 1A-1D, 10, IP, 31A-31I, 32A-32I. In some embodiments, the processes of Figures 31J-31P may be performed using a system for manufacturing a construction material that includes a biologies activation and / or expansion system, such as depicted in Figure 30B and 30C, the incubation system 106 of Figures 1 A-1D, 10, IP and / or the crust formation system of Figure ID, and Figure 30A, and the feed system 108 depicted in Figures 31A-31I.

[0319] Figure 31J depicts a flowchart describing another embodiment of a process for manufacturing a construction material. In step 3122 a metastable shaped structure that includes aggregate particles (e.g., an aggregate blend) and mineralizing enzymes and / or microorganisms is acquired. In step 3124, the shaped structure is submersed into a cementation reagent solution so that the cementation reagent solution infiltrates a plurality of pores in the shaped structure. In step 3126, the mineralizing enzymes or microorganisms to produce a mineral binder in the shaped structure and partially deplete a concentration of cementation reagent in the plurality of pores.

[0320] Figure 3 IK depicts a flowchart describing another embodiment of a process for manufacturing a construction material. In Figure 31K, steps 3132, 3134, and 3136 mirrorsteps 3122, 3124, and 3126 of Figure 31J. Figure 31K adds step 3138. in which an impulse is applied to the shaped structure and / or the cementation reagent solution so that a portion of the cementation reagent solution is displaced from the plurality of pores and is replaced by a portion of the cementation solution that is not / less depleted.

[0321] Figure 3 IL depicts a flowchart describing another embodiment of a process for manufacturing a construction material. In Figure 31L, steps 3142, 3144, and 3146 mirror steps 3122, 3124, and 3126 of Figure 31J. Figure 31L adds step 3148, in which an impulse is applied to displace at least a portion of the depleted cementation reagent solution from the pores, and step 3150 in which fresh cementation solution is caused or allowed to enter the pores.

[0322] Figure 3 IM depicts a flowchart describing another embodiment of a process for manufacturing a construction material. In Figure 31M, steps 3142 and 3144 mirror steps 3122 and 3124 of Figure 31J. Figure 31M adds step 3156 in which the cementation solution is mixed, step 3158 in which a concentration of the cementation reagent or a mineralization reaction byproduct is detected in the bath, and step 3160 in which more cementation reagent is added to the bath at a predetermined time or in response to the detected concentration.

[0323] Figure 3 IN depicts a flowchart describing another embodiment of a process for manufacturing a construction material. In Figure 3 IN, steps 3172 and 3174 mirror steps 3122 and 3124 of Figure 31J. Figure 31N adds step 3176 in which the cementation solution is mixed, step 3158 in which a portion of the cementation reagent solution is transferred to a regeneration system, step 3178 in which more cementation reagent to the portion of the cementation reagent solution in the regeneration system, and step 3180 in which the portion of the cementation reagent solution is returned to the bath.

[0324] Figure 310 depicts a flowchart describing another embodiment of a process for manufacturing a construction material. In step 3182 a metastable shaped structure that includes aggregate particles (e.g., an aggregate blend) and mineralizing enzymes and / or microorganisms is acquired. In step 3184, a first feed solution that includes a cementation reagent is acquired. In step 3186 the first feed solution is applied to the shaped structure with a first set of feed parameters. In step 3188, a second feed solution that includes the cementation reagent is acquired. In step 3190, the second feed solution is applied to the shaped structure using a second set of feed parameters different from the first set of feed parameters.

[0325] Figure 3 IP depicts a flowchart describing another embodiment of a process for manufacturing a construction material. In Figure 3 IP, steps 3192 and 3194 mirror steps 3122 and 3124 of Figure 31 J. Figure 3 IP adds step 3196 in which a portion of the cementation reagent solution is transferred to a regeneration system, step 3198 in which more cementation reagent is added to the portion of the cementation reagent solution in the regeneration system, step 31100 in which the portion of the cementation reagent solution is returned to the bath, and step 31102 in which the previous three steps are repeated until the shaped structure is sufficiently hardened by the action of the mineralizing enzymes and / or microorganisms. In step 31104 in which the shaped structure is removed. In step 31106, the regeneration steps are repeated. In step 31108, a new shaped structure is added.

[0326] Figures 32A-32I depict embodiments of a regeneration system 3009 for regenerating a feed solution used in a process such as the processes described herein. The system may include a first cementation reagent source 3201, an acid source 3003, a recirculation tank 3202, a second cementation reagent source 3203, a computing system 101. The system may also include a pH sensor 124, a concentration sensor 125, and a temperature sensor. The system may also include mixer(s) 3016 and pumps(s) 3012, as shown in Figure 32B. The system may also include a permeable barrier, as shown in Figure 32B, 32C, 32E, and 32F. Which may be part of, or separate from, a byproduct removal system 3108, which may itself be connected to a by-product collection system 3110 (see Figure 31G). The regeneration system 3009 may be configured to allow for convective mixing as shown in Figures 32D-32F, by having a high density’ cementation reagent source 3111 fluidically connect to a top portion of the recirculation tank 3202, and a low density cementation reagent source fluidically connect to a bottom portion of the recirculation tank 3202. This can be achieved in a calcium chloride process since a near-saturated aqueous calcium chloride solution has a greater density than a feed solution. The system may include a nutrient source 3206, a drain valve 3207, an effluent drain 3208, and one or more biologies inactivation system(s) (e g., a hot zone, aUV source, etc ), as shown Figures 32G-32I. Wide arrows show mass flows between and through various sub-systems and elements of the regeneration system, where white arrows show liquid flow (e.g., feed solution, effluent, regenerated feed, acid, cementation reagents, nutrient solution, etc.), black arrows show solid transfer (e.g., precipitated by-products, e.g., precipitated ammonium chloride, e.g., due to mixing of high concentration calcium chloride with spend feed solution containing ammonium chloride, which is induced to precipitate due to the common ion effect, and canbe trapped and collected due to a permeable barrier that allows the regenerated feed solution but not the precipitated solid), black and white arrows show missed solid and liquid transfer (e.g., slurries).

[0327] Figure 32J depicts a flowchart describing another embodiment of a process for manufacturing a construction material including a regeneration step. In embodiments, the process of Figure 32J may be performed in one or more of the systems depicted in Figures 32A-32A, and may be incorporated into the systems and processes depicted in Figures 1 A- 1F, 4J-6C, and 31A-31P.

[0328] In step 3222 a plurality of aggregate particles combined with one or more mineralizing enzymes and / or microorganisms is provided. In step 3234, a feed solution that includes a cementation reagent is provided. In step 3236, the feed solution is applied to the plurality of aggregate particles. In step 3238, the plurality of aggregate particles is cured so that a mineral binder formed by action of the mineralizing enzy mes and / or microorganisms on the cementation reagent consolidates the plurality of aggregate particles, resulting in a depleted feed solution. In step 3240, the depleted feed solution is collected. In step 3242, the collected feed solution is transferred to a regeneration system. In step 3244, more cementation reagent to the depleted feed solution in the regeneration system to form a regenerated feed solution is added. In 3246, the regenerated feed solution is applied to the aggregate particles or to a second plurality of aggregate particles.

[0329] Figure 33 depicts a flowchart describing an embodiment of a process for enhancing a construction material. In step 3302, a construction material that has a network of pores is provided. In step 3304, a first bath including a first reagent solution of a first reagent is provided. In step 3306, a second bath including a second reagent solution of a second reagent is provided. In step 3308, the construction material is submersed into the first bath so that the first reagent solution infills at least a portion of the network of pores. In step 3310, the construction material is submersed into the second bath so that the first reagent and the second reagent react together within the network of pores to form a precipitate that at least partially blocks the network of pores. Alternatively, the first reagent solution and the second reagent solution are applied by one or more spray systems. Or the first step in by submersion and the second step by spraying, or vice versa. The process, or a portion thereof, of Figure 33 may be applied in a feed system such as shown in Figures 1 A- 1L, Figure IN, and Figures 30A and 30D, and Figures 31A-31I. An embodiment of the systems shown in Figures 31 A-3 II with two feed baths may be used. An embodiment of thesystems shown in Figures 31A-31I with one feed baths may be used by draining the feed solution of a first reagent and re-filling with a second.

[0330] Figure 34 depicts a decision flowchart for determining whether to proceed to an inoculation step (e.g.. of a larger tank, or of an aggregate) or continue fermentation (e.g., expansion) in a liquid medium (e g., in a seed flask or bioprocesses tank).

[0331] Figure 35 depicts a pore in a plurality of aggregate particles (e.g., in a shaped structure) and several of the chemical, biological, and material processes that occur therein during urease-based MICP.

[0332] Example - Pre-cast Construction Unit Production System

[0333] In some embodiments, a process is provided for manufacturing pre-cast construction units (e.g., tiles, pavers, bricks, etc.) utilizing a biological (e.g., microbial) agent to produce biocement through one or more enzymatic pathway(s), synthesizing calcium carbonate from carbonate source (e.g., urea, dissolved or dispersed CO2) and calcium substrates supplied to an aggregate mix. Tn some cases, the manufacturing process for producing pre-cast construction units (or units) is designed to produce units of 100-500 mm length, 20-500 mm width, and 5-50 mm thickness. The process includes steps of initial germination, activation, and expansion, initially in a liquid state (or from dried biologies to a liquid seed solution), then in the solid state. The units, once pressed and biologically ready for feeding are then fed by a spray system. Other feeding systems, such as those described herein (e.g., soak, drip, continuous flow, pressure fed, etc.) may also be applied.

[0334] The process when starting from dried microorganisms may be initiated with a liquid germination and / or activation step to expand the concentration and total count of microbial agents in a seed solution to be used in a larger scale liquid activation step. Alternatively, germination and activation and expansion to a desired cell count and biological activity may all be performed in a single bioprocess tank. In some embodiments, one or more of germination, activation, and expansion to a desired cell count and biological activity, may all be performed in the solid state before and / or after pressing to a desired shape. In some embodiments, germination and activation, and expansion to an initial cell count and biological activity, may be performed in one or more liquid bioprocess tanks, then further expansion (which may include some germination and activation) performed in the solid state before and / or after pressing to a desired shape to a second and / or third cell count and / or biological activity.

[0335] Feed media and components may be pasteurized in the bioprocess tank to reduce bioburden. Dry biological agent is added to an intermediate container (which may be provided sterile or sterilized prior to adding the biological agent) of growth medium and incubated for 6-30 hours (e.g., about 5-10 hours, 6-12 hours, 10-20 hours, 12-18 hours, 12- 24 hours, 18-20 hours, 18-24 hours, or 20-24 hours, e.g., about 5, 10, 15, 18, 20, or 24 hours) (or, e.g.. enough time to produce from 1 to 30 population doublings, e.g., 1-4, 1-3. 2-4, 3-5, 4-6. 4-7, 4-8, 4-9. 4-10. 5-7, 5-8, 5-9. 5-10. 6-8, 6-9, 6-10, 7-9. 7-10. 8-10. 8-12. 8-16. 8-24. 8-30, 9-12, 9-15, 9-18, 9-24, 9-30, 10-12, 10-15, 12-16, 12-20, 12-24, 12-30, 13-16, 13-19, 13-25, 14-16, 14-20, 14-24, 14-28, 14-30, 15-17, 15-18, 15-20, 15-24, 15-30, 16-18, 16-19, 16-20, 17-20, 17-30, 18-24, 18-30, 20-30, 22-26, 22-28, or 24-30 population doublings) to produce a seed solution. The sterility of the intermediate container may be greater than that of the subsequent bioprocess tank for larger scale expansion, to reduce bioburden at the earliest stage, and to improve process efficiency at the larger scale. The bioprocess tank is then inoculated with a specified volume of this seed solution to hit a target initial cell concentration (e.g., between about 1 x 103and 1 x 1012CFU per mL, e.g.. between 1 x 105and 1 x 1010CFU per mL, e.g., between 1 x 107and 5 x 109CFU per mL, e.g., between 1 x 108and 5 x 109CFU per mL, e.g., between 1 x 108and 2 x 109CFU per mL, e.g., between 5 x 108and 2 x 109CFU per mL, e.g., between 1 x 108and 1 x 1012CFU per mL, e.g., between 1 x 109and 1 x 1011CFU per mL, e.g., between 1 x 1010and 1 x 1012CFU per mL, or e.g., between 1 x 1010and 1 x 1013CFU per mL). The tank is held at growth conditions during a growth period with pH, temperature, and dissolved oxygen (DO) actively monitored for a time sufficient to expand the cells by approximately 1-10 population doublings, e.g., about 1-3, 2-5, 2-8, 3-5, 3-6, 3-7. 3-8, 3-9, 3-10, 4-6, 4-7, 4-8, 4-9, 4-10, 5- 7, 5-8, 5-9, 5-10, 6-8, 6-9, 6-10, 7-9, 7-10, or 8-10 population doublings.

[0336] After the growth period, the bioprocess tank holds liquid material to be used during a unit expansion step for unit expansion (e.g., having a cell concentration of from about 1 x 104to 1 x 1016CFU per mL, e g., between 1 x 105and 1 x 1010CFU per mL, e.g., between 1 x 107and 5 x 109CFU per mL, e.g., between 1 x 108and 5 x 109CFU per mL, e.g., between 1 x 108and 2 x 109CFU per mL, e.g., between 5 x 108and 2 x 109CFU per mL, e.g., between 1 x 108and 1 x 1012CFU per mL, e.g.. between 1 x 109and 1 x 1011CFU per mL, e.g., between 1 x 109and 1 x 1012CFU per mL, e.g., between 1 x 1010and 1 x 1012CFU per mL, e.g., between 1 x 1010and 1 x 1013CFU per mL, or e.g., between 1 x 1012and 1 x 1014CFU per mL). The bioprocess tank may hold liquid activation material for up to 6hours (e.g., up to 1 hour, up to 2 hours, up to 3 hours, up to 4 hours, up to 5 hours, or up to 6 hours).

[0337] In some embodiments, rather than a two-stage process, the liquid activation may be a single step, either only the "‘intermediate container” step (e.g., a seed tank) or only the bioprocess tank step (i.e., directly adding dried biological agent to the bioprocess tank), prior to the unit expansion step. The volume of inoculum required at the solid-state mixing step may control which pre-mix steps are performed.

[0338] In some embodiments, dried biological agents (e.g., microorganisms, e.g., bacteria) may be directly added to the aggregate in the unit expansion step. Bypassing the liquid activation step(s) may require a greater quantity of dried biological agent (e.g., between about 1 x 103and 1 x 1014CFU per gram of aggregate blend mass, e.g., between 1 x 105and 1 x IO10CFU per gram, e.g., between 1 x 107and 5 x 109CFU per gram, e.g., between 1 x 108and 5 x 109CFU per gram, e.g., between 1 x 108and 2 x 109CFU per gram, e.g.. between 5 x 108and 2 x 109CFU per gram, e.g.. between 1 x 108and 1 x 1012CFU per gram, e.g., between 1 x 109and 1 x 1011CFU per gram, e.g., between 1 x IO10and 1 x 1012CFU per gram, or e.g., between 1 x IO10and 1 x 1014CFU per gram).

[0339] In some cases, the dried biologies are within a powder form or a tablet form. Technical benefits of utilizing a tablet form over a pow der form include a reduction in dust issues, a reduction in material loss due to fine particles adhering to measuring containers, and improved delivery of the correct amount of dried biologies without requiring measuring containers (e.g., a measuring cup). The dried biologies may include microorganisms and nutrients, such as glucose and com steep liquor (CSU) (e.g., solids or dried CSU solutes). The dried biologies may also include urea and a binder, such as Inulin or a prebiotic fiber.

[0340] In some embodiments, the unit expansion step begins with blending various aggregates (e.g., sand, gravel, crushed stone, etc.), and, optionally, additives, and pigment materials, in a mixer where liquid activation material, inoculum, and water are introduced to achieve an aggregate mix with a nominal 1-10% moisture content (e.g., about 1-5%, 2- 5%, 3-5%, 3-6%, 4-5%, 4-6%, 5-6%, 5-7%, 5-10%, 6-7%, 6-8%, 7-8%, 7-9%, 8-10%, or 9- 10%, e.g., about 1%, 2%, 3,%, 4%, 5%, 6%, 7%, 8%, 9%, or 10%) moisture content byweight. The mix is then transferred to a hopper and subsequently fed into a vibrating press to form wet construction units (e.g., tiles (e.g., of dimensions 100-500 mm x 50-250 mm x 6-50 mm, e.g., of dimensions 400 mm x 200 mm x 22 mm), or bricks, or pavers). Followingthe press (e.g., immediately, or, e.g., within 10 minutes to 4 hours, e.g., within about 30 min, 1 hr, 2hrs, 3hrs, or 4hrs), tiles are held in an environmental control chamber for a unit expansion phase of approximately 1-8 hours (e.g., about 1-2 hours, 2-4 hours, 3-5 hours, 4- 6 hours, 5-7 hours, or 6-8 hours, e.g., about 1 hour, 1.5 hours, 2 hours, 2.5 hours, 3 hours, 3.5 hours, 4 hours, 4.5 hours, 5 hours, 5.5 hours, 6 hours, 6.5 hours, 7 hours, 7.5 hours, or 8 hours). The unit expansion process allows for expansion of the cells by 2-10 population doublings, e.g., about 2-5, 2-8, 3-5. 3-6, 3-7, 3-8. 3-9, 3-10, 4-6. 4-7, 4-8, 4-9. 4-10. 5-7, 5- 8, 5-9, 5-10, 6-8, 6-9, 6-10, 7-9, 7-10, or 8-10 population doublings.

[0341] Following unit expansion, the units enter the feed phase of the process. Following pressing into the target geometry, construction units are transferred to a process chamber for a unit incubation phase (where, e.g., germination, activation, expansion, proliferation, fermentation, may occur in the formed (e.g., pressed) solid state) prior to the initiation of the feeding process. Following unit expansion, the process feed sequence is initiated. Rows of units are transferred from the process chamber to a feed station. Feed solution is sprayed over the tiles via a series of spray nozzles operating at a specified flow rate, duty cycle and linear travel speed. After the feeding sequence, rows are transferred to the flex chamber, and a post-feed rest period is initiated.

[0342] The feed process exposes the units to a series of aqueous feed solutions composed of cementation reagent(s) (e.g., a calcium ion source, e.g., calcium chloride) and a carbonate source, e.g., urea, dissolved carbon dioxide, etc.) (and, optionally, nutrients, pH modifiers (e.g., acids, e.g., HC1, sulfuric acid, nitric acid, acetic acid, citric acid, etc.), etc.) followed by post-feed rests. The feeding process begins by removing units from a temperature / humidity controlled environment (e.g., a closed, or partially closed, chamber including a system for modulating moisture (e.g., a source of moisture) system for controlling temperature (e.g., a heat source), which may be separate components or the same component and temperature sensor(s) and hygrometer(s) coupled to the temperature and moisture modulating component(s)) and placing them under a spray system manifold. The spray system performs a senes of passes over the units (or the units move under the spray system), introducing the feed solution that promotes enzymatic (e.g., microbial) production of biocement. After dispensing a target volume of feed solution, the units are moved to a chamber (which may be a climate controlled chamber such as that used for the incubation, if not the same chamber) for a rest period (e.g., of 1 to 8 hours, e.g., about 1-2 hours, 2-4 hours, 3-5 hours, 4-6 hours, 5-7 hours, or 6-8 hours, e.g., about 1 hour, 1.5 hours, 2 hours,2.5 hours, 3 hours, 3.5 hours, 4 hours, 4.5 hours, 5 hours, 5.5 hours, 6 hours, 6.5 hours, 7 hours. 7.5 hours, or 8 hours), e.g., for a total time of 12-96 hours, e.g., from 12-18 hours, 12-24 hours, 18-36 hours, 20-30 hours, 24-36 hours, 24-48 hours, 36-48 hours, 30-40 hours, 30-50 hours, 40-50 hours, 48-60 hours, 48-72 hours, 50-60 hours, 50-70 hours, 50-80 hours, 50-90 hours, or 64-96 hours.

[0343] After completion of a particular number of feeds and rests, the units are rinsed with water, samples are tested for structural integrity and the remaining batch are transferred to a polishing unit operation. Units are fed through a GMM LMS 650 4C-8A automated stone polisher. The units pass through four diamond grind heads to remove excess height and ensure a level surface. They then pass through various polishing heads to achieve the desired surface finish. Upon exiting the machine, the units are partially dried using high velocity fans. Dried, polished tiles are checked for quality control standards, cosmetic quality, and defects.

[0344] Workflow Example:

[0345] Pasteurization

[0346] Liquid Activation and Expansion in Bioprocess Tank

[0347] Example blend formulation table (basis 630 kg.)

[0348] Optimized Aggregate Blends

[0349] In some embodiments, the aggregate densities (e.g., in terms of the void space of an aggregate blend) of the construction materials disclosed herein approach those of ordinary portland cement (OPC). One issue with this is that the manufacturing of OPC does not need to consider bringing reagents into the aggregate matrix, whereas in MICP, flow7pathways are necessary for the cementation reagents to reach the pores.

[0350] Aggregate typically occupies -65-85% by volume in biocement-based construction units. The remaining volume (30-35%) is occupied by other components such as reactive / non-reactive fillers / admixtures / additives, moisture, bacteria, organic material, & nutrients in addition to volumetrically infdl of biocement, latter which is a product resulting from the characteristic biomineralization. A lower infill of biocement (%vol.) can be desirable to keep cost & carbon footprint (quantified in terms of Global Warming Potential (GWP)) as low as possible. Being the dominant phase, aggregate blend optimization is important to: (a) lower the process and product costs, and GWP; (b) indirectly facilitate the overall biomineralization process, e.g., by enhancing contact-point cementation, effective bridging of calcium carbonate crystals, efficient straining of bacterial cells and feed nutrients, enhancing feed liquid and moisture penetration and retention through pore throat refinement; (c) significantly improve the end product mechanical properties (e.g.,compressive & flexural strengths, abrasion resistance) and durability (e.g., fluid absorption, freeze-thaw stability).

[0351] Different aggregate materials and their blends & gradations (e.g., Fuller curves or FHWA-modified Fuller curves) have been optimized to maximize the particle packing density and reduce the associated pore space / pore volume / porosity of pressed units. Surprisingly, both high density and gap graded blends (including higher density gap-graded blends) demonstrated high compressive strength after biocementation in flow-based feeding processes, despite the associated decrease in porosity, particularly open pore porosity, and increase in tortuosity, reducing the available internal surface area available to receive cementation reagents. The associated technical advantages of these blends and gradations are multifold and include:

[0352] (a) reduction in the biocement (%) requirement (by reducing total pore / voids volume) while still satisfying the end product specifications (such as compressive strength, absorption, freeze-thaw performance) in accordance with relevant standards (such as ASTM & EN specifications). For example, when a blend of two naturally sourced aggregates with a starting porosity’ of -35% was optimized for their maximum packing density, it resulted in a final blend porosity of -31% (i.e., 4% reduction in bulk porosity) reducing the need of biocement infill (vol%) by -11% to achieve an identical end mechanical performance.

[0353] (b) associated benefits with reduction in overall cost & carbon footprint (global warming potential-GWP) of the biocement processes and products.

[0354] (c) maximizing the end unit mechanical (for instance, compressive & flexural strengths, abrasion resistance) and durability (for instance, fluid absorption, freeze-thaw) performances. For example, when a blend of two naturally sourced aggregates with a starting porosity' of -35% was optimized for their maximum packing density, to result in a final blend porosity of -31% (i.e., 4% reduction in bulk porosity), the compressive strength of the end unit in the latter case was observed to increase by 27% for a given set of other conditions and parameters. The reduction in the pore volume also reduces the overall water absorption thus improving the freeze-thaw performance of the final unit.

[0355] In some cases, optimized gradation curves may be applied (e.g., to guide in aggregate blends selection) to MICP-based processes such as those described herein. The optimization approach used here for aggregates (e.g.. naturally source aggregates, synthetic aggregates, processed (e.g., crushed) aggregates, etc.) their blends can be extended toaggregates of various geological origin, mineralogy (silicates, calcareous, aluminates, etc.), artificially processed aggregates (e g., crushed quarried aggregates, recycled concrete aggregates, etc.) aggregates produced from dedicated processes including slag aggregates, light weight aggregates, fly ash sintered / pelletized aggregates & other such), and aggregates from waste streams (e.g., mine tailings, bottom ash aggregates, etc.).

[0356] Surprisingly, the particle packing density of aggregate and their blends can be further enhanced by incorporating smaller particle size fractions to fill in larger intra particle pore spaces created by next larger aggregate particles in a sequential fashion, while maintaining sufficient amounts and evenness of biocement formation to bind the aggregate enough to meet the desired properties, despite the associated decrease in porosity, particularly open pore porosity, and increase in tortuosity, reducing the available internal surface area available to receive cementation reagents. These smaller fractions could include reactive or non-reactive filler materials such as limestone, dolomite, pozzolans (such as volcanic ash, fly ash), clay, crushed glass, quarry crusher fines, industrial byproducts (such as municipal solid waste incineration ashes, slag, off-spec fly ashes, etc.). Use of these materials may further reduce the biocement requirements and also reduce the overall GWP of MICP processes.

[0357] Incorporation of fibers into the aggregate blends described herein may allow for even greater packing densities by providing fluid paths which enhance dispersion of fluid into the aggregate matrix. The combination of densely packable aggregate blend with fluid dispersion-enhancing additives such as fibers can allow even greater finished densities, lower porosities, lower GWP, and higher unit strengths (compressive or flexural) than either of these improvements alone. Further, incorporation of fibers can enhance flexural strength.

[0358] Figure 10A depicts representations of different types of aggregate blend gradation types.

[0359] Figure 10B depicts a representation of fluid flowing through an aggregate blend.

[0360] Figure 10C depicts the results of a study of binary aggregate blends with different ratios of Skygge (‘S’ a type of sand) and Thybo (‘T’, a type of gravel) (i.e., of different gradations). The x-axis is %T (therefore the %S is 100-x) and the two y-axis are (left) density (envelope, g / cc) of the blend and (right) water infiltration rate, Ksat (cm / s). The %water retention (%wt) is also shown, but not associated with either axis, the highestvalue was 12.86% and the lowest value was 8.56%. The envelope density of the blends peaks at from about S60T40 to about S40T60.

[0361] Fiber Admixtures in Aggregate Blends

[0362] Flexural or bending strength is a key performance metric construction units, particularly pavers, blocks, tiles, flags, etc. One potential limitation of MICP-based processes, particularly those requiring flow of liquid carrying cementation reagents, is uniformity of cementation, for example when delivery of cementation reagents is from above and driven primarily by gravity. Fiber incorporation can help to distribute the feed solution more evenly throughout the aggregate matrix.

[0363] In an initial experiment, jute fibers showed initial signs of flexural strength improvement. In one experiment, an aggregate mixture including 0.5 wt% of jute fibers achieved a flexural strength of 1.14 MPa at a time when observed flexural strengths were often under 1 MPa. Results of a flask biocementation study indicated that out of the three plant fibers tested (jute, hemp, & flax), flax fibers had slightly higher CaCCh yield, and basalt fibers performed significantly better than all three plant fibers.

[0364] In hand-fed test samples (pressed to 52x22x120mm shape), basalt fibers were added to a baseline formulation (with a ratio of sand (<1 mm diameter) to crushed stone (-1-10 mm diameter) of 55:45) at varying dosages (0.25 wt%, 0.50 wt%, 0.75 wt%). Flexural strength improved at all dosages vs the control, with 0.50 wt% yielding the highest average strength of 1.55 MPa, roughly 20% higher than the control.

[0365] Figures 10D-10E depict results for different formulations that include fiber admixtures.

[0366] Textile reinforced Biocement-Strain Hardened Textile Reinforced Bioconcrete for Improved Flexural Strength, Impact Resistance & Fracture Toughness

[0367] The mechanical performance requirements for pre-cast construction units (e.g., tiles, bricks, flags, pavers, blocks, etc.) may be defined by end unit specifications (e.g., EN 1339) that call for, e.g., a flexural strength greater than or equal to 4 MPa, or may be related to improved impact resistance or enhanced fracture toughness. For achieving some properties, increasing the biocement infill to the microstructure to enhance the end mechanical performance may not be sufficient, or may be prohibitively time or reagent intensive. Reasons for this may include: 1) There is a threshold on the maximum infill thatcan be achieved from a standpoint of pore disconnection that accompanies externally fed biocement-forming processes, thus, increases in the mechanical performance with continued external feeding may plateau at a certain quantity of biocement formation, thus limiting the strength development thereafter irrespective of the incoming feed media. 2) The overall process of increasing the calcium carbonate infill to improve the end unit performance can be relatively slow. 3) Increasing the %CaCOs infill alone may not be the most economical solution, thus restricting wider applicability.

[0368] In some cases, strain hardened biocemented composites can enhance the flexural strength, impact resistance and fracture toughness of end cemented units.

[0369] The composite material will continue to take load post its first crack formation. In other words, the textile reinforced bioconcrete units will have a higher load carrying ability7under flexure or bending stresses (i.e., higher flexural strength). In the absence of textile reinforcement, such stresses can rapidly cause failure of the entire material (e.g., sudden failure / brittle failure, i.e., the units are only as strong as their weakest point). To remedy this, textile reinforced bioconcrete composites were investigated and demonstrated strain hardening behavior. As the embedded textile reinforcement / mesh resists crack propagation and thereby delays the material failure of the unit, the composite material has higher energy absorption capacity prior to failure (higher impact resistance and fracture toughness). This is true in case of both 2D and 3D-textile reinforcement scenarios.

[0370] As a proof of concept, an experiment was conducted using a single textile grid / mesh of natural jute material. With the use of just a single textile grid as demonstrated, the end unit displayed a significant delay in crack propagation, significant extent of strain hardening behavior, and a 32% relative improvement in the end flexural strength of the unit (in third-point bending test). The results are shown in the Figure I OF. These results could find utility in applications such as, e.g., blast resistant structures, paving for outdoor applications where load-bearing capacity is a key metric, e.g., driveways, and even in tiles, pavers, blocks, and flags. Additional embodiments include:

[0371] (1) textile / fiber mesh may be placed in the tension zone of the unit (approx, in the bottom portion, e.g., in the bottom half, bottom third, or bottom quarter, of the unit relative to the intended placement (e.g., for a paver, the bottom is the face in contact with the ground, for a tile, the bottom is the face in contact with the surface to which the tile ismounted). In some cases, to resist tension, the textile may be positioned in the middle of the tension zone or near the middle of the tension zone.

[0372] (2) If more than one mesh is used per unit, then meshes may be spaced in the tension zone.

[0373] Since max tension force gets applied to the bottommost part of the unit per the stress distribution sketch (see Figure 10G), it can be beneficial to place a mesh at the bottom of the mold prior to pouring aggregate into the mold. During pressing, the mesh is embedded into the pressed unit. With cementation, the calcium carbonate minerals can hold the embedded mesh strongly in place. Locating the mesh at bottom of the mold can delay / resist crack initiation (not crack propagation).

[0374] Hydroxide Activated Limestone

[0375] The feedstock requirements / costs of enzyme / microbe-based (e.g., MICP-based) processes can be reduced by incorporating aspects of alkali activated cement into enzyme / microbe-based processes such as those described herein. In particular, alkali (e.g., NaOH, KOH, or MgOH) can be used as an alkaline activator for the fusion of carbonates (e.g., limestone) to impart additional strength to a biocement-based construction material, or to substitute for mineral (e.g.. carbonate) binding formed by mineralizing enzymes or microorganisms with, e.g., MICP feedstocks (e.g., CaCb and urea), thereby reducing feedstock costs (and water usage), numbers of cycles required, total process time, etc. The inclusion of an alkaline binder into our material can fuse a portion of limestone in an uncemented aggregate blend before feed cycles begin. The mineral binder (e.g., calcite) subsequently formed during biocementation is thus provided an easier pathway to bridging the space between aggregate particles thus building strength. The increase in strength can help reduce the number of feeds needed to achieve the same strength as material that does not contain the alkali binder.

[0376] Using the current baseline aggregate mixing and mix-in inoculation method for the NaOH-activated limestone is a challenge due to the high starting pH of the material. However, giving the aggregate time for its pH to decrease could help increase biocompatibility for mix-in units.

[0377] Despite the alkaline condition created by the addition of NaOH, the new material appears biocompatible using topical inoculation, as shown in Figure 10H.

[0378] Units containing 5% limestone (LS) with NaOH appear to have a lower percent change in density than baseline units (2% LS, without NaOH), suggesting that the LS + NaOH material needed less biocement to achieve a higher compressive strength. In addition, all samples showed urease activity: AVG Urea Consumption (mmol / (kg*min)) = A (2% LS) 4.65; B (2% LS + NaOH) 4.71; C (5% LS + NaOH) 2.58. It has thus been shown that, despite the presence of harsh alkali, enzyme or microorganism-based biocementation (e.g., MICP) can be performed on aggregate blends including limestone (e.g., fine limestone, e.g.. ultrafine limestone) that has been treated with alkali (e.g., sodium hydroxide) to improve the overall compressive strength of the resulting construction material. Further improvements may include adding additional buffering agents to the aggregate particles (e.g., with magnesium sulfate) or to the inoculation composition (e.g., citric acid, phosphoric acid, etc.) to further protect microorganisms or enzymes from the alkali, or adding a wash step or neutralization step (e.g., with acetic, hydrochloric, sulfuric, or carbonic acid) to the process to reduce the alkalinity.

[0379] Magnesium hydroxide Carbonation to Enhance Strength, Reduce Cost, and Capture CO2

[0380] Minimally processed minerals that can be carbonated through urea hydrolysis can lead to significant cost savings in biocement / bioconcrete technologies. Alkaline minerals are one such material and present an added benefit of potentially capturing CO2 to form carbonates, reducing the amount of urea required to achieve maximum cementation.

[0381] Brucite, a mineral composed of magnesium hydroxide, has shown potential for use as a biomineralization feedstock. Unlike calcium hydroxide, which is more widely available, brucite is not commonly used as a construction material. All testing has been conducted with reagent grade Mg(OH)2, but minimally processed mineral brucite is also considered as an option.

[0382] The advantages of brucite over calcium hydroxide include lower solubility, which results in a pH that is more tolerable for bacteria (~9.5 vs 12.5 for Ca(OH)2), and a high degree of expansion upon carbonation, which leads to significant porosity reduction. The exact chemistry of brucite carbonation is complex, as there are several magnesium carbonate minerals with varying degrees of hydration and carbonation. Magnesite (MgCCh) is the most stable magnesium carbonate mineral (excluding dolomite, which also requires calcium to form), but metastable hydrated magnesium carbonates are expected to be morereadily formed and can be stable for years or decades in ambient conditions. For this reason, the relevant minerals to our process include nesquehonite (MgCO? 3H2O), dypingite (Mg5(CO?)4(OH)2-5H2O), and hydromagnesite (Mgs(CO3)4(OH)2-4H2O).

[0383] The initial investigation focused on microbial growth in the presence of different alkaline materials (added at 5% w / w w.r.t. S55T45). Microbial growth was assessed in terms of urease activity after 6 hours of solid-state incubation. Little to no growth was observed for calcium hydroxide or magnesium oxide, but magnesium hydroxide showed minimal inhibition of microbial growth (subsequent tests have consistently shown a -50% reduction in urease activity compared to baseline samples).

[0384] A MiniFlex tile (52x120x22mm) was also prepared, and received one feed of IM urea + 2g / L yeast extract after the 6hr pre-feed incubation. The sample was then incubated for 72 hours in 20% CO2 before being rinsed and dried at 45 °C. The mechanical performance of this sample was impressive, with 2.7 MPa flexural strength and 3,677 psi compressive strength, despite only receiving one feed and a relatively short CO2 exposure. SEM analysis of this sample indicated an abundance of dypingite and / or hydromagnesite, which present similar morphologies.

[0385] Follow-up experiments investigated extended feeding & curing regimes using the same formulation and CO2 concentration. 7 feeds without CO2 curing resulted in average strengths of 2.3 MPa and 1,906 psi, organism-free controls with 7 days of CO2 curing yielded 1.4 MPa and 4.304 psi. and 7 feeds followed by 7 days of CO2 curing yielded 1.7 MPa and 4,105 psi. While these experiments did not yield improved flexural performance, they provided insight into the effect of CO2 curing on mechanical performance, especially compressive strength.

[0386] In these experiments, Mg(OH)2 dosage was held constant at 5% based on unit cost estimates, but higher dosages may also be advantageous. Other options that are considered include alternate aggregate blends, combinations with other additives (limestone, fibers, etc.), and alternate fermentation / incubation approaches to enhance microbial growth & urealysis rates.

[0387] In some embodiments, the magnesium hydroxide is produced in situ by the reaction of magnesium sulfate added to the aggregate blend and hydroxide ions that are extracted from another aggregate component (e g., blast furnace slag, pozzolan, recycled OPC concrete, etc.) upon hydration with water or feed solution.

[0388] Freeze-Thaw Enhancement Studies

[0389] The biocemented structures (construction materials, bioconcrete), such as construction units (e.g., tiles, pavers, blocks, large format precast elements, etc., which may also be referred to generally as biostructures herein) can benefit from material property enhancing additives, for example, to improve freeze-thaw cycling resistance, or flexural strength in wet conditions. For example, units that are fed by gravity or diffusion-based mechanisms may result in finished units with relatively open pore structures, which can allow water to penetrate.

[0390] Investigations have been performed, and several additives have been found to be compatible with the biostructures described herein and can be implemented at commercial scale. These include additives that resist water penetration, that interfere with ice crystal formation, or that incorporate micropores (e.g., with microspheres).

[0391] PEG-PVA Copolymer as Ice Recrystallization Inhibitor for Freeze-Thaw Improvement

[0392] The use of a PEG-PVA copolymer to improve the freeze-thaw resistance for construction materials produced with biocementation technologies (e.g., using the methods described herein). The copolymer interacts with water in pores to reduce the size of ice crystals that form during freeze-thaw cycles.

[0393] The copolymer may be added into a biocementation process at the beginning as a ‘'mix-in” component with the aggregate. Additional moisture added with the copolymer into the aggregate may improve workability. Alternatively, a soluble version of the polymer may be dissolved in an appropriate non-aqueous solvent and added to the finished construction unit. The “mix-in” approach has the advantage of guaranteeing highly homogeneous incorporation of the copolymer into the aggregate matrix and pore network, maximizing the benefit. Further, a copolymer that is handled as a solid can be made sufficiently long to be insoluble in water, thereby ensuring that the polymer remains in the construction material and is not washed out.

[0394] There is only limited research on using PEG-PVA as a freeze-thaw solution in traditional concrete (see, e.g.. Frazier et al. (Frazier et al., Cell Reports Physical Science 1, 100060, June 24, 2020), who explored the use of PEG-PVA as an alternative to airentraining admixtures for reducing freeze-thaw damage in traditional concrete, and found that PEG-PVA at wt. 0.021% of OPC paste showed no damage after 30 freeze-thaw cyclesin comparison PVA and PEG, which showed damage). Traditional concrete is very different from biocement (e.g., MICP)-based materials, for example, having different pore matrix structure and different chemistry. Challenges with implementing this, or similar copolymers, include determining the proper dosage since available dosage information is based on systems using OPC, chemical compatibility with the calcite-based binder, and biological compatibility’ with the enzymes and microorganisms, particularly when mixing the copolymer into the aggregate at the start of the process (thereby ensuring that the biological elements are exposed to the copolymer for the entire bioconcrete formation process).

[0395] Enhanced Fertilizer

[0396] Direct application of chemical nitrogen fertilizers can be detrimental to some crops, causing, e.g., fertilizer bum, or over-provision of certain nutrients, leading to metabolic harm and even crop death. Salinity increase and soil acidification are also known problems arising from the use of single chemical nitrogen sources (e.g.. urea pellets or dry ammonia). Further, nitrogen is not the only need that crop plants have. For example, many plants require chloride (e.g., date palm, coconut, etc.) or calcium to thrive. Finally, the soil microbiome has its own needs (such as accessible forms of organic carbon, e.g., sugars, acids, amino acids, proteins, etc.), which may not be met by simple chemical fertilizers, and which can be challenging to provide at scale.

[0397] MICP processes (e.g., urease-based processes) such as those described herein produce compositions of ammonium chloride and metabolic by-products / waste products (e.g., amino acids, proteins, peptides, organic acids, complex sugar molecules, etc.), i.e., ammonium chloride (or another ammonium salt, e.g., ammonium sulfate, e.g., prepared as described herein) enhanced with nutrients produced by the microorganisms, which can provide a beneficially enhanced nitrogen fertilizer, once concentrated. The combination of an ammonium salt (e.g., ammonium chloride or ammonium sulfate) and nutrients nourishes not just the crop plant, but the soil microbiome holistically.

[0398] Such an enhanced fertilizer may also include unreacted urea and calcium chloride, providing further benefits. For example, the mixture of urea and ammonium will provide nitrogen to the crop soil at different rates, providing both immediate and long-term benefit. The calcium chloride can provide these nutrients where lacking, and, in the case ofcalcium, contribute to soil deacidification by conversion to calcium carbonate in the soil, either by native organisms, or by spores present in the fertilizer.

[0399] Viable spores in the fertilizer can further benefit the soil by deacidifying the soil by urealysis, which produces ammonia, which acts to both raise the pH and to provide nitrogen to the plants, and by calcite formation. In the presence of calcium ions from the fertilizer composition, the urea-producing microorganisms, once germinated from the spores, will naturally buffer the soil to a pH close to neutral. Further, by generating calcareous material in the soil, mineral retention, water retention, and organic matter retention are all enhanced.

[0400] Concentrated ammonium fertilizer blends produced with ammonium and other chemicals produced by mineralizing (e.g., urease-forming or carbonic anhydraseproducing) microorganisms thus have the potential to provide high value-added fertilizers to enhance or remediate soil conditions and improve crop yields.

[0401] Stripping of HCl Vapor from the Waste Stream via Acidification

[0402] In some embodiments, the disclosed technologies may include recyclingHCl, e.g., to use to produce more calcium chloride from calcium carbonate (e.g., limestone), with a strong acid, e.g., sulfuric acid. Water primarily goes with the ammonium sulfate product. Carbon dioxide produced by reaction of HCl can be captured and converted into urea, or captured and forced into a feed solution, e.g., a feed solution containing carbonic anhydrase and / or in a sealed or partially sealed system.

[0403] Some embodiments may include stripping ammonia and acid from effluent and reusing for a complete recycle.

[0404] Some embodiments may include using the ammonium sulfate produced to make a concentrated ammonium fertilizer, such as by the methods described herein.

[0405] Producing Calcium Chloride from Waste Ammonium Chloride with Hydroxide

[0406] In some embodiments, systems and methods include elements that recycle ammonium chloride by pH modulation with a strong base, e.g., hydroxide ions, e.g., from calcium hydroxide, blast furnace slag, recycled cement, etc., to evolve ammonia gas and produce calcium chloride: NH4CI (aq) + CaOH (aq) NH? (g) + CaCh (aq) + H2O (1).

[0407] In some embodiments, the calcium chloride is originally derived from calcium carbonate. In some embodiments, the resulting calcium chloride is re-used as a calcium source in methods described herein. In some embodiments, the resulting calcium chloride is further treated (e.g., filtered, sterilized, etc.) before being used as a calcium source in a method described herein.

[0408] Producing Calcium Chloride from Waste Ammonium Chloride and Limestone

[0409] In some embodiments, systems and methods include elements that recycle ammonium chloride to CaCh. In some embodiments, the calcium chloride is originally derived from calcium carbonate. For example, a process that upcycles a waste stream of ammonium chloride (NH4CI) into a useful feedstock of calcium chloride (CaCh), while recovering or valorizing the by-product ammonia (NH3) and carbon dioxide (CO2). This concept draws inspiration from the cyclic chemistry of the Solvay process, where ammonia and CO2 shuttle between different steps, yet is streamlined to target CaCh as the principal product.

[0410] The Core Chemical Scheme1. Dissolve Calcium Carbonate via CO2In a dedicated reactor, limestone (CaCCh) is contacted with water and dissolved CO2 to form soluble calcium bicarbonate [CaQTCChh]. This step is comparable to creating an ‘'acidic” bicarbonate solution without adding strong mineral acids.2. React with Ammonium Chloride• The bicarbonate solution is treated with NH4Q (the waste stream), which transforms the calcium ions into CaCh.• The nitrogen component ends up as ammonium bicarbonate (NH4HCO3).3. Thermally Decompose Ammonium Bicarbonate• Mild heating (e g., 70-100 °C) drives off NH3 and CO2:• 2 NH4HCO3 — > 2 NH3+ 2 C02+ 2 H2OThe recovered NH3 and CO2 can be captured for sale, recycling back into the process, or further conversion. In short: CaCO? + 2 NH4C1 — > CaCb+2NH3 + 2 CO2 + 2 H2O (With some of the CO2 and NH3optionally recycled to solubilize more CaCCh.)

[0411] Processes may further include coupling NH3 + CO2 in a urea reactor, making an additional high-demand feedstock (urea) while continuously regenerating the CaCU solution.

[0412] Processes may integrate multiple resource loops — "circular’7strategies that valorize multiple waste streams simultaneously.

[0413] Systems to perform effluent recycling and / or revalorization may include a dissolution reactor which can operate with moderate CO2 pressure or continuous sparging to produce Ca(HCO3)2 quickly and efficiently. Processes may further include systems for solid-liquid separation such that any undissolved CaCCh or inert solids can be screened out before the bicarbonate solution moves on. Advantageously, recovery systems and methods such as described herein may involve thermal decomposition at low (e.g., 70-100 °C) temperatures, in contrast to the than the 900 °C calcination required in Solvay’s lime kiln process. This milder step still requires energy but can tap waste heat or other on-site heat sources for further efficiencies. Processes such as described herein may afford resource efficiency by leveraging abundant limestone (CaCOs) without the need for strong mineral acids. Systems such as those described herein are highly scalable and flexible: The process can be set up at smaller scales (where NH4Q is generated as a sidestream) or scaled up with integrated ammonia / CCh capture. Further, such methods and systems may be integrated into existing CO2 generation or capture systems, affording synergistic efficiencies and valorizations, for example if a plant already has CO2 and steam streams (e.g., from fermentation or power generation), this synergy can reduce the net carbon footprint.

[0414] By adapting the concept of a “cyclic” ammonia-CCh process — akin to Solvay — but focusing on the valuable production of calcium chloride, a waste NFUC1 stream is transformed into a marketable feedstock. Along the way, ammonia and CO2 are recovered, both of which can be sold, converted into ammonium sulfate, or upgraded into urea for sale or re-use in processes described herein (adding additional flexibility).

[0415] Comparison to the Solvay Process

[0416] The Solvay process produces soda ash (Na2COs) from NaCl and CaCCh, with CaCh as a by-product. Ammonia is circulated to capture and release CO2, while a high- temperature kiln provides fresh CO2 (and quicklime). Our concept follows a similar ammonia-CO2 cycle — yet aims directly at CaCh using a waste NH4CI stream. We skip the lime kiln and avoid generating large volumes of CaCh ‘‘waste.’' Instead, the desired product is CaCb, and we recover NHa and CO2 in a mild thermal step. Thereby reducing embodied carbon emissions in finished products.

[0417] At least one embodiment of the disclosed technology includes providing a shaped structure that includes mineralizing (e.g., urease producing or carbonic anhydraseproducing) microorganisms and a plurality of aggregate particles; acquiring a feed solution that includes a cementation reagent; setting a single-use feed volume for the feed solution based on a pore volume for the plurality of aggregate particles; and applying the single-use feed volume of the feed solution to the shaped structure during one or more single-use feed cycles, the applying the single-use feed volume of the feed solution causes calcium carbonate to bind at least two particles of the plurality of aggregate particles.

[0418] At least one embodiment of the disclosed technology includes providing a shaped structure that includes mineralizing (e.g., urease producing or carbonic anhydraseproducing) microorganisms and a plurality of aggregate particles; acquiring a feed solution that includes a cementation reagent and has a molarity greater than 700mM of the cementation reagent; and applying the feed solution to the shaped structure during one or more feed cycles, the applying the feed solution to the shaped structure causes calcium carbonate to bind at least two particles of the plurality of aggregate particles, the applying the feed solution to the plurality of aggregate particles includes applying single-use feed cycles or soaking the plurality of aggregate particles in a bath of the feed solution.

[0419] At least one embodiment of the disclosed technology includes acquiring a plurality of aggregate particles that have been formed into a shape; acquiring a feed solution that has a pH less than 6 and includes a cementation reagent; and applying the feed solution to the plurality of aggregate particles during one or more feed cycles, the applying the feed solution to the plurality of aggregate particles causes calcium carbonate to bind at least two particles of the plurality of aggregate particles.

[0420] At least one embodiment of the disclosed technology includes forming a shaped structure that includes mineralizing (e.g., urease producing or carbonic anhydrase-producing) microorganisms and a plurality of aggregate particles, wherein the forming includes allowing the shaped structure to harden to a sufficiently meta-stable state subsequent or prior to inclusion of the urease producing microorganisms; acquiring a first feed solution that includes a cementation reagent; subsequent to allowing the shaped structure to harden, submersing the shaped structure in a bath of the first feed solution, the submersing the shaped structure in the bath of the first feed solution causes calcium carbonate to bind at least two particles of the plurality of aggregate particles; and removing the shaped structure from the bath of the first feed solution after at least 20% of the cementation reagent has been consumed.

[0421] At least one embodiment of the disclosed technology includes generating a mixture of an inoculation composition comprising a plurality of mineralizing (e.g., urease producing) microorganisms and / or spores thereof and a plurality’ of aggregate particles; forming a shaped structure using the mixture of the inoculation composition and the plurality of aggregate particles; incubating the mixture of the mineralizing (e g., urease producing) microorganisms subsequent to forming the shaped structure; acquiring a feed solution that includes a cementation reagent; applying the feed solution to the shaped structure during one or more feed cycles, the applying the feed solution causes calcium carbonate to bind at least two particles of the plurality of aggregate particles.

[0422] At least one embodiment of the disclosed technology includes providing a shaped structure that includes mineralizing (e.g., urease producing) microorganisms and a plurality of aggregate particles; acquiring a first feed solution that includes a cementation reagent; applying the first feed solution to the shaped structure using a first set of feed cycles with a first set of feed parameters, the applying the first feed solution causes calcium carbonate to bind at least two particles of the plurality of aggregate particles; acquiring a second feed solution that includes the cementation reagent; and applying the second feed solution to the shaped structure using a second set of feed cycles with a second set of feed parameters different from the first set of feed parameters.

[0423] At least one embodiment of the disclosed technology includes acquiring a shaped structure that includes mineralizing (e.g.. urease producing) microorganisms and a plurality of aggregate particles; acquiring a feed solution that includes a cementation reagent; determining an ambient temperature and ambient humidity7; determining an estimated infiltration rate and an estimated evaporation rate; determining a first set of feed parameters for a first set of feed cycles based on the ambient temperature, the ambient humidity, theestimated infiltration rate, or the estimated evaporation rate; and applying the feed solution to the shaped structure using the first set of feed cycles with the first set of feed parameters, the applying the feed solution causes calcium carbonate to bind at least two particles of the plurality of aggregate particles.

[0424] In some embodiments, a feed solution has an ionic strength greater than 2.1 moles per liter (e.g., from 4 to 20 moles per liter, from 2 to 10 moles per liter, from 3 to 10 moles per liter, from 2 to 8 moles per liter, from 2 to 6 moles per liter, from 2 to 5 moles per liter, from 2 to 4 moles per liter, from 2 to 3 moles per liter, from 2.5 to 5 moles per liter, from 2.5 to 4.5 moles per liter, from 2.5 to 3.5 moles per liter, from 3 to 4 moles per liter, from 3 to 6 moles per liter, from 3 to 8 moles per liter, from 3 to 10 moles per liter, from 3 to 12 moles per liter, from 3 to 15 moles per liter, from 3 to 20 moles per liter, from 3.5 to 4.5 moles per liter, from 3.5 to 5 moles per liter, from 4 to 6 moles per liter, from 4 to 8 moles per liter, from 4 to 10 moles per liter, from 4 to 12 moles per liter, from 4 to 15 moles per liter, from 4 to 20 moles per liter, e.g., about 2.0, 2. 1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 7.0, 8.0, 9.0, 10.0, 12.0. 14.0, 15.0, 16.0, 18.0, or 20.0 moles per liter, e.g., greater than 2.0, 2.1. 2.3, 2.4, 2.5. 2.6, 2.7, 2.8, 2.9, 3.0, 3.5, 4.0, 4.5, 5.0, 6.0, 8.0, 10.0, or 15 moles per liter). In some embodiments, a feed solution has an osmotic pressure greater than 1.7 MPa at 25 degrees Celsius (e.g., greater than 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13. 14. 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, or 50 MPa at 25 degrees Celsius, e.g., greater than 2, 3, 4. 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15. 16. 17, 18, 19, 20, 25, 30, 35, 40, 45, or 50 MPa at 35 degrees Celsius, e.g., greater than 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, or 50 MPa at 40 degrees Celsius, e.g., from 2 to 60 MPa at 25 degrees Celsius, from 2 to 60 MPa at 35 degrees Celsius, from 2 to 60 MPa at 40 degrees Celsius, from 5 to 15 60 MPa at 35 degrees Celsius, from 10 to 20 MPa at 35 degrees Celsius, from 15 to 45 MPa at 25 degrees Celsius, e.g., about 5, 10, 15, 20, 25, 30, 35, 40, 45, or 50 MPa at 25 degrees Celsius), has an osmotic pressure greater than 3MPa at 25 degrees Celsius, has an osmotic pressure greater than 6.7MPa at 25 degrees Celsius, has an osmotic pressure greater than 10 MPa at 25 degrees Celsius, or has an osmotic pressure greater than 15 MPa at 25 degrees Celsius. In some embodiments, a feed solution has an osmolarity of at least 3 Osm / L (e.g., about 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, 14, 14.5, 15, 16, 17, 18, 19, or 20 Osm / L, e g., between 3 and 5, between 3 and 10, between 3 and 12,between 3 and 15, between 3 and 20, between 4 and 6, between 4 and 8, between 4 and 12, between 4 and 20, between 5 and 10, between 5 and 15. between 5 and 20, between 7 and 12, between 7 and 15, between 7 and 21, between 8 and 16, between 8 and 22, between 9 and 12, between 9 and 18, between 9 and 21, between 10 and 15, between 10 and 20, between 10 and 25, between 11 and 17, between 11 and 21, between 12 and 16, between 12 and 22, between 15 and 20, or between 15 and 25 Osm / L, e.g., greater than 3, 5. 6, 8, 9, 10, 12, 15, or 20 Osm / L). In some embodiments, a feed solution has an osmolarity of at least Osm / L (e.g., greater than 3, 5, 6, 8, 9, 10, 12, 15, or 20 Osm / L). In some embodiments, a feed solution has a delivered pH of from about 11 to about 1 (e.g., from 8 to about 1, e.g., from 9 to about 2, e.g., from 8 to about 3. e.g., an acidic pH, e.g., about 7, 6, 5, 4, 3, 2. or 1, e.g.. from 7 to 2, e.g., from 6 to 3, e.g., from 4 to 2. In embodiments, an osmolarity increases in the pores during incubation / curing. In embodiments, a pH in the pores after feed delivery ranges from 2 to 10, e.g.., from 3 to 10, e.g., from 3 to 7, e.g., reaching a final pH of about 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, or 10. In some cases, it has been discovered that in the conditions of the methods described herein, the microorganisms are able to function at in feed solutions having an order of magnitude greater osmotic pressure than would be expected to be tolerable without at least inducing sporulation (and thus stopping biocementation) or, more likely, inducing widespread cell death to the population of microorganisms. For example, the feed solution may be hyperosmotic relative to (e.g., highly hyperosmotic, e.g., 10-100x greater than) an osmotic pressure of the fluid inside the cells of the microorganisms. In some embodiments, the feed solution comprises from 0.01 to 2 M of ammonium chloride.

[0425] The concentration of calcium ions in a cementation reagent solution (e.g., a feed solution, bath, etc.) may independently be about 0.1 M to about 4 M. The concentration of calcium ions in a cementation reagent solution may independently be about 0.1 M to about 0.2 M, about 0.1 M to about 0.3 M, about 0.1 M to about 0.4 M, about 0.1 M to about 0.5 M, about 0. 1 M to about 0.7 M, about 0.1 M to about 0.9 M, about 0. 1 M to about 1 M, about 0.1 M to about 1.2 M, about 0.1 M to about 1.5 M, about 0.1 M to about 2 M, about 0.1 M to about 3 M, about 0.2 M to about 0.3 M, about 0.2 M to about 0.4 M. about 0.2 M to about 0.5 M. about 0.2 M to about 0.7 M, about 0.2 M to about 0.9 M, about 0.2 M to about 1 M, about 0.2 M to about 1.2 M, about 0.2 M to about 1.5 M, about 0.2 M to about 2 M, about 0.2 M to about 3 M, about 0.3 M to about 0.4 M, about 0.3 M to about 0.5 M, about 0.3 M to about 0.7 M, about 0.3 M to about 0.9 M, about 0.3 M to about 1 M, about0.3 M to about 1.2 M, about 0.3 M to about 1.5 M. about 0.3 M to about 2 M. about 0.3 M to about 3 M, about 0.4 M to about 0.5 M, about 0.4 M to about 0.7 M, about 0.4 M to about 0.9 M, about 0.4 M to about 1 M, about 0.4 M to about 1.2 M, about 0.4 M to about 1.5 M, about 0.4 M to about 2 M, about 0.4 M to about 3 M, about 0.5 M to about 0.7 M, about 0.5 M to about 0.9 M, about 0.5 M to about 1 M, about 0.5 M to about 1.2 M, about 0.5 M to about 1.5 M, about 0.5 M to about 2 M. about 0.5 M to about 3 M, about 0.7 M to about 0.9 M, about 0.7 M to about 1 M. about 0.7 M to about 1.2 M, about 0.7 M to about 1.5 M, about 0.7 M to about 2 M, about 0.7 M to about 3 M, about 0.9 M to about 1 M, about 0.9 M to about 1.2 M, about 0.9 M to about 1.5 M, about 0.9 M to about 2 M, about 0.9 M to about 3 M, about 1 M to about 1.2 M, about 1 M to about 1.5 M, about 1 M to about 2 M, about 1 M to about 3 M, about 1.2 M to about 1.5 M, about 1.2 M to about 2 M, about 1.2 M to about 3 M, about 1.5 M to about 2 M, about 1.5 M to about 3 M, about 2 M to about 3 M, about 2 M to about 4 M, about 2.5 M to about 4 M, or about 3 M to about 4 M. The concentration of calcium ions in a cementation reagent solution may independently be about 0.1 M, about 0.2 M, about 0.3 M, about 0.4 M. about 0.5 M, about 0.7 M, about 0.9 M, about 1 M, about 1.2 M. about 1.5 M, about 2 M, or about 3 M. The concentration of calcium ions in a cementation reagent solution may independently be at least about 0.1 M, about 0.2 M, about 0.3 M, about 0.4 M, about 0.5 M, about 0.7 M, about 0.9 M, about 1 M, about 1.2 M, about 1.5 M, about 2 M. or about 3 M.

[0426] The concentration of urea in a cementation reagent solution (e.g., a feed solution, bath, etc.) may independently be about 0.1 M to about 4 M. The concentration of urea in a cementation reagent solution may independently be about 0. 1 M to about 0.2 M, about 0. 1 M to about 0.3 M, about 0.1 M to about 0.4 M, about 0.1 M to about 0.5 M, about 0.1 M to about 0.7 M, about 0.1 M to about 0.9 M, about 0.1 M to about 1 M, about 0.1 M to about 1.2 M, about 0.1 M to about 1.5 M, about 0.1 M to about 2 M, about 0.1 M to about 3 M, about 0.2 M to about 0.3 M, about 0.2 M to about 0.4 M. about 0.2 M to about 0.5 M, about 0.2 M to about 0.7 M, about 0.2 M to about 0.9 M, about 0.2 M to about 1 M, about 0.2 M to about 1.2 M, about 0.2 M to about 1.5 M, about 0.2 M to about 2 M, about 0.2 M to about 3 M, about 0.3 M to about 0.4 M. about 0.3 M to about 0.5 M, about 0.3 M to about 0.7 M, about 0.3 M to about 0.9 M, about 0.3 M to about 1 M, about 0.3 M to about 1.2 M, about 0.3 M to about 1.5 M, about 0.3 M to about 2 M, about 0.3 M to about 3 M, about 0.4 M to about 0.5 M, about 0.4 M to about 0.7 M, about 0.4 M to about 0.9 M, about 0.4 M to about 1 M, about 0.4 M to about 1.2 M, about 0.4 M to about 1.5 M, about 0.4 M to about 2 M,about 0.4 M to about 3 M, about 0.5 M to about 0.7 M, about 0.5 M to about 0.9 M, about 0.5 M to about 1 M, about 0.5 M to about 1.2 M, about 0.5 M to about 1.5 M. about 0.5 M to about 2 M, about 0.5 M to about 3 M, about 0.7 M to about 0.9 M, about 0.7 M to about 1 M, about 0.7 M to about 1.2 M, about 0.7 M to about 1.5 M, about 0.7 M to about 2 M, about 0.7 M to about 3 M, about 0.9 M to about 1 M, about 0.9 M to about 1.2 M, about 0.9 M to about 1.5 M, about 0.9 M to about 2 M, about 0.9 M to about 3 M, about 1 M to about1.2 M, about 1 M to about 1.5 M, about 1 M to about 2 M, about 1 M to about 3 M, about1.2 M to about 1.5 M, about 1.2 M to about 2 M, about 1.2 M to about 3 M, about 1.5 M to about 2 M, about 1.5 M to about 3 M, about 2 M to about 3 M, about 2 M to about 4 M, about 2.5 M to about 4 M, or about 3 M to about 4 M. The concentration of urea in a cementation reagent solution may independently be about 0. 1 M. about 0.2 M. about 0.3 M, about 0.4 M, about 0.5 M, about 0.7 M, about 0.9 M, about 1 M, about 1.2 M, about 1.5 M, about 2 M, about 2.5 M, about 3 M, about 3.5 M, or about 4 M. The concentration of urea in a cementation reagent solution may independently be at least about 0.1 M, about 0.2 M, about 0.3 M, about 0.4 M, about 0.5 M, about 0.7 M. about 0.9 M, about 1 M, about 1.2 M, about 1.5 M. about 2 M, about 2.5 M, about 3 M. about 3.5 M. or about 4 M. The concentration of urea in a cementation reagent solution may independently be at most about 0.2 M, about 0.3 M, about 0.4 M, about 0.5 M, about 0.7 M, about 0.9 M, about 1 M, about1.2 M, about 1.5 M, about 2 M. or about 4 M.

[0427] The flowcharts and block diagrams in the figures provide illustrations of the architecture, functionality, and operation of possible implementations of systems, methods, and computer program products according to various aspects of the disclosed technology7. In this regard, each step in a flowchart may correspond with a program module or portion of computer program code, which may include one or more computer-executable instructions for implementing the specified functionality. In some implementations, the functionality noted within a step may occur out of the order noted in the figures. For example, two steps shown in succession may, in fact, be executed substantially concurrently, or the steps may sometimes be executed in the reverse order, depending upon the functionality involved. In some implementations, steps may be omitted and other steps added without departing from the spirit and scope of the present subject matter.

[0428] In some implementations, the functionality noted within a step may be implemented using hardware, software, or a combination of hardware and software. Asexamples, the hardware may include microcontrollers, microprocessors, field programmable gate arrays (FPGAs), and electronic circuitry.

[0429] In some embodiments, a bioconcrete construction material produced by the methods or systems described herein has a compressive strength of about 900 psi to about6.800 psi. In some embodiments, a bioconcrete construction material has a compressive strength of about 900 psi to about 1,000 psi, about 900 psi to about 1,100 psi, about 900 psi to about 1,200 psi, about 900 psi to about 1,300 psi, about 900 psi to about 1,400 psi, about 900 psi to about 1,600 psi, about 900 psi to about 1,800 psi, about 900 psi to about 2,000 psi, about 900 psi to about 2,500 psi, about 900 psi to about 3,000 psi, about 900 psi to about3.500 psi, about 1,000 psi to about 1,100 psi, about 1,000 psi to about 1,200 psi, about 1,000 psi to about 1,300 psi, about 1,000 psi to about 1,400 psi, about 1,000 psi to about 1,600 psi, about 1.000 psi to about 1,800 psi. about 1,000 psi to about 2,000 psi, about 1,000 psi to about 2.500 psi. about 1,000 psi to about 3,000 psi. about 1,000 psi to about 3,500 psi, about1.800 psi to about 2,000 psi, about 1,800 psi to about 2,500 psi, about 1,800 psi to about 3,000 psi, about 1,800 psi to about 3,500 psi, about 2,000 psi to about 2,500 psi, about 2,000 psi to about 3,000 psi, about 2,000 psi to about 3,500 psi, about 2,500 psi to about 3,000 psi, about 2.500 psi to about 3,500 psi, or about 3,000 psi to about 3.500 psi, about 1,100 psi to about 4,200 psi, about 2,100 psi to about 4,300 psi, about 2,100 psi to about 5,400 psi, about 2,100 psi to about 5,600 psi, about 2,100 psi to about 4,800 psi, about 2,100 psi to about6.800 psi, about 1,100 psi to about 6,500 psi, about 4,100 psi to about 6,000 psi, about 5,100 psi to about 6.500 psi, about 4,200 psi to about 6,800 psi, or about 5,200 psi to about 6,800 psi. In some embodiments, a bioconcrete construction material has a compressive strength of about 1,000 psi, about 1, 200 psi, about 1,500 psi, about 2,000 psi, about 2,200 psi, about2.500 psi, about 3,000 psi, about 3,200 psi, about 3,500 psi, about 4,000 psi, about 4,200 psi, about 4,500 psi, about 5,000 psi, about 5,200 psi, about 5,500 psi, about 6,000 psi, about 6,200 psi, or about 6.500 psi. In some embodiments, a bioconcrete construction material has a compressive strength of at least 900 psi, 1 ,000 psi, 2,000 psi, 3,000 psi, 4,000 psi, 5,00 psi, 6,000 psi, or 6,800 psi.

[0430] In some embodiments, any construction material described herein (e.g., made with any of the methods or systems described herein) has a flexural strength of from 1 to 10 MPa, (e.g., from 1 to 5 MPa, from 2 to 6 MPa, from 2 to 10 MPa, from 3 to 9 MPa, from 4 to 8 MPa, from 3 to 10 MPa, from 5 to 10 Mpa, from 3 to 10 MPa, from 4 to 10MPa, from 5 to 7 MPa, from 8 to 10 MPa, or from 9 to 10 MPa. In some embodiments, any construction material described herein (e.g., made with any of the methods or systems described herein) has a flexural strength of at least 1 MPa, at least 2 MPa, at least 3 MPa, at least 4 MPa, 5 MPa, at least 6 MPa, at least 7 MPa, at least 8 MPa, at least 9 MPa, or at least 10 MPa. In some embodiments, any construction material described herein (e.g.. made with any of the methods or systems described herein) has a flexural strength of about 1 MPa. about 2 MPa, about 3 MPa, about 4 MPa, 5 MPa, about 6 MPa, about 7 MPa, about 8 MPa, about 9 MPa, about 10 MPa, about 11 MPa, about 12 MPa, about 13 MPa, about 14 MPa, 15 MPa, about 16 MPa. about 17 MPa. about 18 MPa. about 19 MPa, or about 20 MPa.

[0431] In some embodiments, any construction material described herein remains undamaged after 5 cycles of an EN 14617-5 freeze / thaw test. In some embodiments, any construction material described herein remains undamaged after 10 cycles of an EN 14617- 5 freeze / thaw7test. In some embodiments, any construction material described herein remains undamaged after 25 cycles of an EN 14617-5 freeze / thaw test. In some embodiments, any construction material described herein remains undamaged after 30 cycles of an EN 14617-5 freeze / thaw test. In some embodiments, any construction material described herein, or construction material produced according to methods described herein, retains a flexural strength of at least 80% of its original flexural strength after 25 cycles of an EN 14617 freeze / thaw test (e.g., has a KMF25 of at least 80%, 85%, 90%, or 95%). In some embodiments, any construction material descnbed herein, or construction material produced according to methods described herein, retains a flexural strength of at least 50% of its original flexural strength after 25 cycles of an EN 14617 freeze / thaw7test. In some embodiments, any construction material described herein, or construction material produced according to methods described herein, has a water absorbance of less than 5% (e.g., less than 4%, 3%, 2% or 1%, e.g., from 1% to 4%, 2% to 4%, or 3% to 4%, e.g., about 1%, 2%, 2.5%, 3%, or 3.5%) after 25 cycles of an EN 14617 freeze / thaw7test. In some embodiments, any construction material described herein demonstrates less than 1.5 kg / m2of mass loss after 28 cycles of an EN 1339 freeze / thaw test.

[0432] In some embodiments, any construction material described herein has a compressive strength of at least about 3000 psi (e.g.. at least 4000 psi. or at least 6000 psi), and / or wherein the construction material remains undamaged after 25 cycles of an EN 14617-5 freeze / thaw7test. In some embodiments, any construction material described hereinhas a compressive strength of at least about 3000 psi (e.g., at least 4000 psi, or at least 6000 psi), and wherein the construction material remains undamaged after 25 cycles of an EN 14617-5 freeze / thaw test. In some embodiments, any construction material described herein has a compressive strength of at least about 3000 psi (e.g., at least 4000 psi, or at least 6000 psi), or wherein the construction material remains undamaged after 25 cycles of an EN 14617-5 freeze / thaw test. In some embodiments, any construction material described herein has a compressive strength of at least about 3000 psi (e.g., at least 4000 psi, or at least 6000 psi). In some embodiments, any construction material described herein remains undamaged after 25 cycles of an EN 14617-5 freeze / thaw test.

[0433] In some embodiments, blocks (e.g., concrete masonry units, e.g., hollow concrete blocks) produced by the methods described herein, from compositions described herein, or having compositions described herein meet or exceed the standards of ASTM C55 or ASTM C90 (or corresponding EN or ISO standards) for loadbearing applications.

[0434] In some embodiments, any construction material described herein (e.g., made with any of the methods or systems described herein) has a finished density of from about 2 g / cc to about 3 g / cc, for example, from about 1.8 g / cc to about 1.9 g / cc, from about 1.9 g / cc to about 2.0 g / cc, from about 2.0 g / cc to about 2.1 g / cc, from about 2.1 g / cc to about 2.2 g / cc, from about 2.2 g / cc to about 2.3 g / cc. from about 2.3 g / cc to about 2.4 g / cc, from about 2.4 g / cc to about 2.5 g / cc. from about 2.5 g / cc to about 2.6 g / cc. or from about 2.6 g / cc to about 2.7 g / cc, from about 1.8 g / cc to about 2.3 g / cc, from about 1.9 g / cc to about 2.4 g / cc, from about 2.0 g / cc to about 2.7 g / cc, from about 2.1 g / cc to about 2.2 g / cc. In some embodiments, the average finished density is from about 2.0 g / cc to about 2.1 g / cc. In some embodiments, the average finished density is from about 2.1 g / cc to about 2.3 g / cc. In some embodiments, the average finished density is from about 2.2 g / cc to about 2.4 g / cc. In some embodiments, the average finished density is about 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2. or 3.3 g / cc. In some embodiments, the average finished density is greater than 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4. 2.5, 2.6, or 2.7 g / cc.

[0435] Construction materials (e.g., bricks, blocks, slabs, tiles, pavers, etc.) described herein, or produced using methods or compositions described herein, may be bonded together, or bonded to other construction materials using, e.g., cement mortar (e.g., OPC mortar), biocement mortar, carbon-dioxide absorbing mortars, non-OPC cementitious material mortars (e.g., using pozzolans, geopolymers, magnesia, brucite, etc ), to build larger structures, e.g., walls or other kinds of barrier or support structure. In someembodiments, the mortar has a lower GWP than traditional OPC mortars. In some embodiments, the mortar is carbon neutral or carbon-sequestering. In some cases, construction materials are attached together or directly connected using one or more adhesive layers to form larger structures (e.g., walls, columns, or support structures). In one example, the adhesive layer comprises cement mortar.

[0436] Improving Freeze-Thaw Properties by Intentional Precipitation of Metal Carbonate(s) in a Pore Structure

[0437] Bioconcrete construction materials such as those described herein may be further enhanced, by at least partially blocking their pore networks with chemically precipitated metal (e.g., calcium) carbonate cry stals. For example, a metal carbonate (e.g., calcium carbonate) may be induced to crash-out in the pores of a finished construction material (e.g., a tile, paver, block, etc.), in order to block them to retard water ingress. For example, by combining ammonium carbonate and calcium chloride in the pores at high concentrations. Such methods may include deliberately creating a large amount of nonbridging mineral binder crystals.

[0438] Some recent discoveries have highlighted an outsized importance of the role of water absorption in the freeze-thaw properties of bioconcrete construction materials.

[0439] Biocement-based processes (such as those described herein) may produce materials that pass strength parameters for certain outdoor application, due to the high strength and quality of the biocement formed, but may not pass freeze-thaw tests (such as mass loss, water absorption, change in flexural strength, etc.) due to having an inherently high porosity relative to their strength. Adding additional carbonate, which may not form as strong a binder as the biocement mineral binder (e.g., they crash-out / rapidly precipitated crystals rather than slowly grow large, strong, highly interlocking crystals), but can still blocks pores, for example, by reducing the available volume for water ingress by occupying pore volume, or by closing pore throats, making whole pores inaccessible to water ingress. Such precipitates can thus reduce water absorption and allow construction material that may otherwise pass outdoor strength but fail one or more freeze-thaw tests to become a fully freeze-thaw-passing construction material.

[0440] In some cases, ammonium carbonate and calcium chloride can react together in the pores of construction materials to produce calcium carbonate. This calcium carbonate produced in the pores reduces water absorption and thus improves freeze-thaw properties. Insome embodiments, other soluble metal salts may be used, such as salts of magnesium, sodium, potassium, aluminum, copper, zinc, etc. In some embodiments, the salts are chlorides. In some embodiments, a mixture of metal ions may be provided, e.g., a mixture of calcium and magnesium salts. In some embodiments, the metal ions produce carbonates that have a higher molar volume than calcite (e.g., vaterite, aragonite, dolomite, huntite, malachite, azurite, etc.).

[0441] In some embodiments of the methods described herein, calcium cementation reagents may be fully or partially replaced (or partially to fully, e.g., over multiple feeds) with other metal ion (e.g., ions of magnesium, sodium, potassium, aluminum, copper, zinc, etc.) cementation reagents in order to create other metal carbonates from the action of the mineralizing enzymes or microorganisms. In some embodiments, these carbonates have a greater molar volume than calcite e.g., vaterite, aragonite, dolomite, huntite, malachite, azurite, etc ).

[0442] In some cases, while preventing crash-out (e.g., formation of calcium carbonate on or near the surface of a construction material) is desirable to prevent clogging during the initial manufacturing stages of a construction material, intentionally inducing crash-out on or near the surface of a finished construction material (but pre-polished or coated) or during the final manufacturing stages of the construction material improves freeze-thaw properties of the finished construction material. For example, by inducing the conditions that some have previously highlighted as to be avoided to prevent crash-out (as described herein, or as described in WO2024186775A1), or by imposing at a late stage in the process an opposite or inverse of a crash-out-preventing condition (as described herein, or as described in WO2024186775 Al) that is imposed at an earlier stage in the process.

[0443] The following embodiments describe a non-exhaustive list of methods of performing this material enhancement.

[0444] In some embodiments, a finished (but pre-polished or coated) construction material (e.g., a tile, paver, or block) is dipped into a first bath of highly concentrated (e.g., near-saturated) calcium chloride. The construction material is then removed from the first bath and added to a bath of high concentration (e g., near-saturated) ammonium carbonate solution. Alternatively, the first bath is ammonium carbonate and the second bath is calcium chloride. In some embodiments, the construction material is allowed to drain between baths, e g., to leave only what volume of the first bath solution that is retained due to surfacetension. In other embodiments, the construction material is immediately submersed in the second bath, without allowing drainage (e.g., while the pores retain a majority of the volume of fluid from the first bath that inflowed therein while in the first bath. In some embodiments, the construction material is repeatedly transferred between the first and second bath. In some embodiments, the construction material is allowed to fully dry between switching from one bath to the other, such that crystals of ammonium carbonate or calcium chloride form within the pores of the construction material.

[0445] In some cases, it is possible to take advantage of different solubility vs temperature relationships of the starting materials. For example, calcium chloride becomes more soluble at higher temperatures, while ammonium carbonate becomes less soluble with increasing temperature. For example, the saturation concentration of ammonium carbonate at 15 °C is about Ig / mL, but at 20 °C it is only about 0.25 g / ml. The saturation concentration of calcium chloride depends on its hydration state, but showed an increase in solubility with temperature, for example, the hexahydrate can vary from about 0.65 g / ml at 10 °C to >1 g / ml at 30 °C. Methods described herein may be done with baths set to temperatures that are aligned for high solubility of each reagent, and thus non-aligned for the other. For example, in embodiments where ammonium carbonate is precipitated in an inter-bath dry ing step (after submersion in an ammonium carbonate bath at, e.g., 10-15 °C, then added to a warm (e.g., about 30-40 °C) solution of calcium chloride, the suppressed solubility of the ammonium carbonate crystals in the warm second solution can allow time for calcium chloride to more fully penetrate into the pore network of the construction material (e.g., reaching pores nearer the center of the material). As the water cools and more ammonium carbonate dissolves, it will find calcium chloride to react with. Such a method can reduce risk of too much precipitation at the surface blocking internal pores from receiving the same treatment. For example, preventing the enhancement from being confined to surface regions of the construction material.

[0446] Other technical benefits of these methods include the fact that, since ammonium carbonate decomposes to its constituent gases at about 60 °C, so the reaction can use an excess of ammonium carbonate to ensure full consumption of calcium chloride and bake off any excess avoid trapping large amounts of ammonium carbonate in the final pore network. In other embodiments, excess ammonium carbonate may be permitted to remain in the pores, so that it can react with any unreacted metal ions from the aggregate orprocess, thereby adding further carbonate crystals over time. Another technical benefit of inducing precipitation of mineral crystals (e.g., crystals of a mineral binder, e.g., a carbonate, e.g., including calcium carbonate) is enhanced polish-ability, as the precipitated crystals in the pores and at the surface of a structure can resist damage during honing and polishing.

[0447] Figure 11A depicts one embodiment of a non-porous pallet (on the left) without holes or channels and a porous pallet (on the right) with a uniform distribution ...

Claims

1. CLAIMS1. A method for producing a construction material, comprising: acquiring or generating a mixture of an inoculation composition comprising a plurality of mineralizing microorganisms and / or spores thereof and a plurality of aggregate particles; forming a shaped structure using the mixture of the inoculation composition and the plurality of aggregate particles; incubating the mixture of the mineralizing microorganisms subsequent to forming the shaped structure; acquiring a feed solution that includes a cementation reagent; applying the feed solution to the shaped structure during one or more feed cycles, the applying the feed solution causes a mineral binder to bind at least two particles of the plurality of aggregate particles.

2. The method of claim 1, wherein: the incubating the mixture causes an increase in a number of cells of the mineralizing microorganisms (e.g.. urease producing microorganisms).

3. The method of claim 1, further comprising: regulating a temperature of the shaped structure while incubating the mixture of the mineralizing microorganisms (e.g., urease producing microorganisms) such that the temperature of the shaped structure is between 18 and 38 degrees Celsius, e.g., between 28 and 38 degrees Celsius, between 28 and 34 degrees (e.g., about 30 degrees) or, e.g., between 34 and 38 degrees Celsius, e.g., between 35 and 36 Celsius (e.g., about 35 degrees Celsius), or e.g., between 33 and 38 degrees Celsius.

4. The method of claim 1, further comprising: forming a crust layer within the shaped structure prior to applying the feed solution to the shaped structure.

5. The method of claim 4, wherein: the shaped structure has an average moisture content of between 2% and 10% (e.g.,between 2% and 6%, between 3% and 7 %, between 5% and 10%. between 4% and 6%) and / or the crust layer has a moisture content of less than 6% by weight.

6. The method of claim 4, wherein: the forming the crust layer includes drying the shaped structure using environmental conditions that maintain an average moisture content of the shaped structure that is between 3 and 6% by weight.

7. The method of claim 1, further comprising: arranging the shaped structure in a climate controlled environment while incubating the mixture of the mineralizing microorganisms (e.g., urease producing microorganisms) and regulating a humidity of a climate controlled environment while incubating the mixture of the urease producing microorganisms.

8. The method of claim 1, further comprising: positioning the shaped structure in a climate controlled environment while incubating the mixture of the mineralizing microorganisms (e.g., urease producing microorganisms) and regulating a humidity and a temperature of a climate controlled environment while incubating the mixture of the mineralizing microorganisms (e.g., urease producing microorganisms) to maintain a viable moisture content of the shaped structure that is between 3 and 6% by weight.

9. The method of claim 1, wherein: the generating the mixture further comprises adding nutrients (e.g., dried nutrients, or a liquid, or a slurry, e.g., a com steep liquor-based nutrient, e.g., low or non-soluble nutrients, such as com steep solids) to the aggregate.

10. The method of claim 1, further comprising: providing sufficient time and heat to achieve a threshold urease activity (e.g., 1-8. 2-6, 2-10, 3-4, 4-5, 5-10, 6-15, 8-12, 10-20, 9-14, or 3-8 mmol L'1min1).

11. The method of claim 1, further comprising: detecting a specific urease activity, or a metric that corresponds to the specific urease activity (e.g., a threshold rate of ammonia or carbon dioxide production), in the shapedstructure of 1 to 20 (e.g., about 5 to 15, e.g., about 10) micromoles of urea hydrolyzed per billion cells per minute; and / or the incubating comprises expanding the population of mineralizing microorganisms until the specific urease activity in the shaped structure is from 1 to 20 (e.g., about 5 to 15, e.g., about 10) micromoles of urea hydrolyzed per billion cells per minute and / or the urease activity in the shaped structure is from 0.5 to 20 mmol kg’1min’1(millimoles per kilogram of aggregate particles per minute) (e.g., 1-8, 2-6. 2-10. 3-4, 3-8. 4-5, 4-6, 5-10, 6-15, 8-12. 10-20, 9-14, or mmol kg’1min’115-20, e.g., 3-8 mmol kg’1min’1for certain strains of Sporosarcina); and / or the incubating comprises causes or allows expansion of the cells by 1-10 population doublings (e.g., about 2-5, 2-8, 3-5. 3-6, 3-7, 3-8, 3-9, 3-10, 4-6. 4-7, 4-8, 4-9. 4-10. 5-7, 5- 8, 5-9, 5-10, 6-8, 6-9, 6-10, 7-9, 7-10, or 8-10 population doublings).

12. The method of claim 1, wherein: the shaped structure is formed (e.g., pressed or molded) into a shape corresponding to a final construction material, e.g.. a tile structure, a brick structure, a rectangular prism structure, a masonry unit, a paver, or a three-dimensional structure (e.g., a block with one or more cavities (e.g., rectangular or cylindrical cavities, e.g., extending partially or fully through a cross section of the block), or any other structure described herein).

13. A system for producing a construction material, comprising: a biomanufacturing system configured to: generate a mixture of an inoculation composition comprising a plurality of mineralizing microorganisms and / or spores thereof and a plurality of aggregate particles; form a shaped structure using the mixture of the inoculation composition and the plurality' of aggregate particles; incubate the mixture of the mineralizing microorganisms subsequent to forming the shaped structure; acquire a feed solution that includes a cementation reagent; apply the feed solution to the shaped structure during one or more feed cycles, the applying the feed solution causes a mineral binder to bind at least two particles of the plurality of aggregate particles.

14. A system for manufacturing a construction material, comprising: a storage device for storing instructions that, when executed, cause the system to perform operations comprising: generating a mixture of an inoculation composition comprising a plurality of mineralizing microorganisms and / or spores thereof and a plurality of aggregate particles; forming a shaped structure using the mixture of the inoculation composition and the plurality of aggregate particles; incubating the mixture of the mineralizing microorganisms subsequent to forming the shaped structure; acquiring a feed solution that includes a cementation reagent; applying the feed solution to the shaped structure during one or more feed cycles, the applying the feed solution causes a mineral binder to bind at least two particles of the plurality' of aggregate particles.

15. The system of claim 14. further comprising one or more of: a biologies activation / expansion system (3001); a mixing tank (102) (e g., a cement mixing tank); a staging tank (3005) (e.g., coupled to a system of pumps); an incubation system (106) (e.g., including one or more heat sources, sensors, humidifiers, fans, filters, etc.); a computing system (101); one or more pumps (3012); a conveyor system (120) (e.g., including a conveyor belt, a carousel, robotic arm conveyor, a monorail conveyor, an overhead conveyor, and / or a crane); a first cementation reagent source (e.g., a first cementation reagent tank, e.g., ; a second cementation reagent source; an acid source (e.g., an acid tank, e.g., configured to store hydrochloric or sulfuric acid, e.g., coupled to a system of pipes and pumps, e.g., connecting to the staging tank); a pH sensor; a concentration sensor (e.g., an EC sensor); a temperature sensor (e.g., a thermocouple, a thermistor, or a semiconductor-based sensor); a humidity sensor;a heating and / or cooling system (e.g., including one or more fans, heating elements, cooling elements, etc.); a gas source (e.g., an oxygen source, a carbon dioxide source, etc., e.g., fluidly coupled to the biologies activation and / or expansion tank and / or the feed system); a clean-in-place system (e.g., including a tank configured to store one or more CIP reagents, a system of pumps and pipes to connect to, e.g., the staging tank, the feed system, an effluent system, etc.); an effluent system (3008) (e g., including one or more filters (e g., microfilters), a biologies inactivation system, an effluent tank, an effluent drain valve, etc.); or a regeneration system (3009) (e.g., including one or more permeable barriers, pumps, mixers, etc.).

16. A method for producing a construction material, comprising: providing a shaped structure that includes mineralizing enzymes or microorganisms and a plurality of aggregate particles; acquiring a feed solution that includes a cementation reagent; setting a single-use feed volume for the feed solution based on a pore volume for the plurality' of aggregate particles; and applying the single-use feed volume of the feed solution to the shaped structure during one or more single-use feed cycles, the applying the single-use feed volume of the feed solution causes calcium carbonate to bind at least two particles of the plurality of aggregate particles.

17. The method of claim 16, further comprising: identifying one or more dimensions of the shaped structure; and estimating the pore volume for the plurality of aggregate particles using the one or more dimensions of the shaped structure.

18. The method of claim 16, wherein: the shaped structure comprises one of a tile structure, a brick structure, a rectangular prism structure, a masonry unit, a paver, or a three-dimensional structure (e.g., a block with one or more cavities (e.g., rectangular or cylindrical cavities, e.g., extending partially or fully through a cross section of the block).

19. The method of claim 16, further comprising: adjusting the single-use feed volume for the feed solution based on an amount of feed solution that was not absorbed by the plurality of aggregate particles during the one ormore single-use feed cycles, the adjusting the single-use feed volume for the feed solution includes increasing the single-use feed volume for the feed solution.

20. The method of claim 16, further comprising: adjusting the single-use feed volume for the feed solution based on an amount of feed solution that was not absorbed by the plurality of aggregate particles during the one or more single-use feed cy cles, the adjusting the single-use feed volume for the feed solution includes decreasing the single-use feed volume for the feed solution.

21. The method of claim 16, wherein: the setting the single-use feed volume for the feed solution includes setting the feed volume for the feed solution based on a composition of the plurality of aggregate particles.

22. The method of claim 16, further comprising: adjusting a temperature of the single-use feed solution, or a temperature of a climate controlled environment, or both, in order to maintain an average temperature of between 28 and 38 degrees Celsius, e.g., between 28 and 34 degrees (e.g., about 30 degrees) or, e.g., between 34 and 38 degrees Celsius, e.g., between 35 and 36 Celsius (e.g., about 35 degrees Celsius)in the shaped structure.

23. The method of claim 16, wherein: the feed solution does not include nutrients for the mineralizing microorganisms (e.g., urease producing microorganisms).

24. The method of claim 16, wherein: the feed solution comprises nutrients that are substantially soluble in the feed solution.

25. The method of claim 16, wherein: the plurality of aggregate particles includes a gelling agent (e.g., cornstarch, agar, tapioca starch, potato starch, wheat starch, inulin, microcrystalline cellulose, guar gum, sodium alginate, pectin, gelatin, etc.). In some embodiments, the gelling agent is a high surface energy (e.g., having a low water contact angle (e.g., <60 degrees, e.g., 1-50 degrees, 1-40 degrees, 1-30 degrees. 1-20 degrees. 10-40 degrees. 10-30 degrees, etc.) gelling agent, e.g.. alginate.

26. The method of claim 16, wherein: the applying the single-use feed volume of the feed solution to the plurality ofaggregate particles during the one or more single-use feed cycles causes the plurality of aggregate particles to adhere into a construction material or to form a three-dimensional solid structure.

27. The method of claim 16, further comprising: combining the mineralizing enzymes or microorganisms (e.g., urease or urease producing microorganisms) and the plurality of aggregate particles prior to forming the shaped structure.

28. The method of claim 16, wherein: each of the one or more single-use feed cycles comprise a deposition period and a holding / curing period.

29. The method of claim 16, further comprising: applying one or more feed pulses, each having a pulse volume, a pulse pressure, and a pulse duration during a first feed cycle of the one or more single-use feed cycles.

30. A method for producing a construction material, comprising: forming a plurality of aggregate particles into a shape; acquiring a feed solution that includes a cementation reagent; applying the feed solution to the plurality of aggregate particles using one or more single-use feed cycles; and curing the plurality of aggregate particles while the plurality of aggregate particles is within a climate controlled environment with a curing temperature and a curing humidity.

31. A system for producing a construction material, comprising: a feed system configured to: hold a shaped structure that includes one or more mineralizing enzymes or microorganisms (e.g., urease producing microorganisms) and a plurality of aggregate particles; acquire a feed solution that includes a cementation reagent: set a single-use feed volume for the feed solution based on a pore volume for the plurality of aggregate particles; and apply the single-use feed volume of the feed solution to the shaped structure during one or more single-use feed cycles, the applying the single-use feed volume of the feed solution causes calcium carbonate to bind at least two particles of the plurality of aggregate particles.

32. A system for manufacturing a construction material, comprising: a storage device for storing instructions that, when executed, cause the system to perform operations comprising: acquiring a feed solution that includes a cementation reagent; setting a single-use feed volume for the feed solution based on a pore volume for a shaped structure comprising a plurality of aggregate particles and one or more mineralizing enzymes or microorganisms; and applying the single-use feed volume of the feed solution to the shaped structure during one or more single-use feed cycles, the applying the single-use feed volume of the feed solution causes calcium carbonate to bind at least two particles of the plurality of aggregate particles.

33. The system of claim 32, further comprising one or more of: a biologies activation system (3001); a mixing tank (102) (e.g., a cement mixing tank); a staging tank (3005) (e.g., coupled to a system of pumps); an incubation system (106) (e.g., including one or more heat sources, sensors, humidifiers, fans, filters, etc.); a computing system (101); one or more pumps (3012); a conveyor system (120) (e.g., including a conveyor belt, a carousel, robotic arm conveyor, a monorail conveyor, an overhead conveyor, and / or a crane); a first cementation reagent source (e.g., a first cementation reagent tank, e.g., : a second cementation reagent source; an acid source (e.g., an acid tank, e.g., configured to store hydrochloric or sulfuric acid, e.g., coupled to a system of pipes and pumps, e.g., connecting to the staging tank); a pH sensor; a concentration sensor (e.g., an EC sensor); a temperature sensor (e.g., a thermocouple, a thermistor, or a semiconductor-based sensor); a humidity’ sensor; a heating and / or cooling system (e.g., including one or more fans, heating elements, cooling elements, etc.); a gas source (e.g., an oxygen source, a carbon dioxide source, etc., e.g., fluidly coupled to the biologies activation and / or expansion tank and / or the feed system); a clean-in-place system (e.g., including a tank configured to store one or more CIP reagents, a system of pumps and pipes to connect to, e g., the staging tank, the feedsystem, an effluent system, etc.); an effluent system (3008) (e.g.. including one or more filters (e.g., microfilters), a biologies inactivation system, an effluent tank, an effluent drain valve, etc.); or a regeneration system (3009) (e.g.. including one or more permeable barriers, pumps, mixers, etc.).

34. A method for producing a construction material, comprising: forming one or more shaped structures, each comprising a plurality of aggregate particles and one or more mineralizing enzymes or microorganisms; acquiring a first feed solution that includes an amount of a cementation reagent; submersing the shaped structure in a bath of the first feed solution so that a first portion of the first feed solution enters a plurality of pores in the shaped structure, allowing a mineral binder to form from the cementation reagent which binds at least two particles of the plurality of aggregate particles; and removing the shaped structure from the bath of the first feed solution after a portion of the amount of cementation reagent has been converted to the mineral binder.

35. The method of claim 34, wherein the portion of the amount of cementation reagent consumed is at least 10% (e.g., at least 15%, 20%, 25%, 30%, 35%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%, e.g., up to 100%, e.g., from 10% to 100%, from 10% to 95%, from 10% to 75%, from 10% to 50%, from 20% to 95%, from 20% to 100%, from 20% to 95%, from 20% to 90%, from 20% to 85%, from 20% to 80%, from 20% to 75%, from 20% to 70%, from 20% to 65%, from 20% to 60%, from 20% to 55%, from 20% to 50%, from 20% to 45%, from 45% to 100%, from 45% to 95%, from 45% to 90%, from 45% to 85%, from 45% to 80%, from 45% to 75%, from 45% to 70%, from 45% to 65%, from 45% to 60%, from 50% to 100%, from 50% to 75%, or from 50% to 60%. e.g., about 20%, 30%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%) of the amount of cementation reagent.

36. The method of claim 34, further comprising allowing sufficient time for the portion of the amount of cementation reagent consumed to be at least 10% (e.g.. at least 15%. 20%. 25%, 30%, 35%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%, e.g., up to 100%, e.g., from 10% to 100%, from 10% to 95%, from 10% to 75%, from 10% to 50%, from 20% to 95%, from 20% to 100%, from 20% to 95%, from 20% to 90%, from 20% to85%, from 20% to 80%, from 20% to 75%, from 20% to 70%, from 20% to 65%, from 20% to 60%, from 20% to 55%, from 20% to 50%, from 20% to 45%, from 45% to 100%. from45% to 95%, from 45% to 90%, from 45% to 85%, from 45% to 80%, from 45% to 75%, from 45% to 70%, from 45% to 65%, from 45% to 60%, from 50% to 100%, from 50% to 75%, or from 50% to 60%, e.g., about 20%, 30%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%) of the amount of cementation reagent.

37. The method of claim 34, wherein the forming comprises allowing the one or more shaped structures to harden to a meta-stable state subsequent or prior to inclusion of the mineralizing enzymes or microorganisms (the meta-stable state may have a final shaped structure (e.g., construction material, e.g., a tile structure, a brick structure, a rectangular prism structure, a masonry unit, a paver, or a three-dimensional structure (e.g., a block with one or more cavities (e.g., rectangular or cylindrical cavities, e.g., extending partially or fully through a cross section of the block), or any other structure described herein).

38. The method of claim 37. wherein the meta-stable state is sufficiently stable to withstand submersion into the bath.

39. The method of claim 34, wherein a volume of first feed solution in the bath is from 0.9 to 5 times (e.g., from 0.9 to 1 times. 0.9 to 1.1 times, 0.9 to 1.2 times. 0.9 to 1.3 times, 0.9 to 1.4 times, 0.9 to 1.5 times, 0.9 to 1.6 times, 0.9 to 1.7 times, 0.9 to 1.8 times, 0.9 to 1.9 times, 0.9 to 2 times, 0.9 to 2. 1 times, 0.9 to 2.2 times, 0.9 to 2.3 times, 0.9 to 2.4 times, 0.9 to 2.5 times, 0.9 to 2.6 times, 0.9 to 2.7 times, 0.9 to 2.8 times, 0.9 to 2.9 times, 0.9 to 3 times, 0.9 to 3.1 times, 0.9 to 3.2 times. 0.9 to 3.3 times, 0.9 to 3.4 times, 0.9 to 3.5 times, 0.9 to 3.6 times, 0.9 to 3.7 times, 0.9 to 3.8 times, 0.9 to 3.9 times, 0.9 to 4 times, 0.9 to 4.1 times, 0.9 to 4.2 times, 0.9 to 4.3 times, 0.9 to 4.4 times, 0.9 to 4.5 times, 0.9 to 4.6 times, 0.9 to 4.7 times, 0.9 to 4.8 times, 0.9 to 4. 9 times, 0.9 to 5 times, e.g., 1 to 1.1 times, 1 to 1.2 times, 1 to 1.3 times, 1 to 1.4 times, 1 to 1.5 times, 1 to 1.6 times, 1 to 1.7 times, 1 to 1.8 times, 1 to 1.9 times. 1 to 2 times, 1 to 2. 1 times, 1 to 2.2 times. 1 to 2.3 times. 1 to 2.4 times, 1 to 2.5 times, 1 to 2.6 times, 1 to 2.7 times, 1 to 2.8 times, 1 to 2.9 times, 1 to 3 times, 1 to 3.1 times, 1 to 3.2 times, 1 to 3.3 times, 1 to 3.4 times, 1 to 3.5 times, 1 to 3.6 times, 1 to 3.7 times, 1 to 3.8 times, 1 to 3.9 times, 1 to 4 times. 1 to 4.1 times, 1 to 4.2 times, 1 to 4.3 times. 1 to 4.4 times, 1 to 4.5 times, 1 to 4.6 times, 1 to 4.7 times, 1 to 4.8 times. 1 to 4. 9 times, or 1 to 5 times, e.g., 1.5 to 1.6 times, 1.5 to 1.8 times, 1.5 to 2 times, 1.5 to 2.2 times,1.5 to 2.4 times. 1.5 to 2.6 times, 1.5 to 2.8 times, 1.5 to 3 times, 1.5 to 3.2 times, 1.5 to 3.4 times, 1.5 to 3.6 times, 1.5 to 3.8 times. 1.5 to 4 times, 1.5 to 4.2 times, 1.5 to 4.4 times, 1.5 to 4.6 times, 1.5 to 4.8 times, or 1.5 to 5 times, e.g., 1.8 to 2.2 times, 2 to 2.2 times, 2 to 2.4 times, 2 to 2.6 times, 2 to 2.8 times, 2 to 3 times, 2 to 3.2 times, 2 to 3.4 times, 2 to 3.6 times, 2 to 3.8 times, 2 to 4 times, 2 to 4.2 times, 2 to 4.4 times, 2 to 4.6 times, 2 to 4.8 times, or 2 to 5 times, e.g., 2.5 to 3 times, 2.5 to 3.5 times, 2.5 to 4 times. 2.5 to 4.5 times, or 2.5 to 5 times, e.g.. 3 to 3.3 times. 2.5 to 3.6 times, 2.5 to 3.9 times, 2.5 to 4.2 times. 2.5 to 4.5 times, 2.5 to 4.8 times, 2.5 to 5 times, e.g., 4 to 4.1 times, 4 to 4.2 times, 4 to 4.3 times, 4 to 4.4 times, 4 to 4.5 times, 4 to 4.6 times, 4 to 4.7 times, 4 to 4.8 times, 4 to 4.9 times, or 4 to 5 times) a total envelope volume of the one or more shaped structures.

40. The method of claim 34, further comprising displacing at least a portion of a volume of liquid from a plurality of pores within the one or more shaped structures and allowing a portion of the first feed solution re-fill the pores with a portion of the first feed solution in the bath that was previously outside of the plurality of pores.

41. The method of claim 34, wherein the displacing comprises applying an impulse to either the one or more shaped structures or to the first feed solution in the bath.

42. The method of claim 34. further comprising re-submersing the one or more shaped structures into the bath after allowing at least a portion of a volume of liquid in a plurality of pores in the shaped structure to drain from the plurality of pores.

43. The method of claim 34, further comprising repeatedly re-submersing the one or more shaped structures into the bath or repeatedly displacing the portion of the volume of liquid from the plurality of pores within the one or more shaped structures.

44. The method of claim 34, further comprising: acquiring a second feed solution and submersing the one or more shaped structures in a bath of the second feed solution subsequent to removing the one or more shaped structures from the bath of the first feed solution.

45. The method of claim 34, wherein:the submersing the one or more shaped structures in a bath of the first feed solution includes regulating a temperature of the first feed solution while the one or more shaped structures is in the bath of the first feed solution.

46. The method of claim 34, wherein: the submersing the one or more shaped structures in the bath of the first feed solution includes circulating the first feed solution while the one or more shaped structures is in the bath of the first feed solution.

47. The method of claim 34, wherein: the submersing the one or more shaped structures in the bath of the first feed solution includes agitating the first feed solution while the one or more shaped structures is / are in the bath of the first feed solution.

48. The method of claim 34, further comprising: performing pre-feed inoculation to the one or more shaped structures prior to the submersing in the bath of the first feed solution.

49. The method of claim 34, wherein: the submersing the one or more shaped structures includes sealing the one or more shaped structures within a pressurized environment while in the bath of the first feed solution.

50. The method of claim 34. further comprising adding an additional amount of cementation reagent after the portion of the amount of cementation reagent has been converted to the mineral binder.

51. A system for producing a construction material, comprising: a feed system configured to: provide a first feed solution that includes an amount of a cementation reagent to one or more shaped structures, each comprising a plurality of aggregate particles and one or more mineralizing enzymes or microorganisms by submersing the shaped structure in a bath of the first feed solution so that a first portion of the first feed solution enters a plurality of pores in the shaped structure, allowing a mineral binderto form from the cementation reagent which binds at least two particles of the plurality of aggregate particles.

52. A system for manufacturing a construction material, comprising: a storage device for storing instructions that, when executed, cause the system to perform operations comprising: providing a first feed solution that includes an amount of a cementation reagent to one or more shaped structures, each comprising a plurality of aggregate particles and one or more mineralizing enzy mes or microorganisms by submersing the shaped structure in a bath of the first feed solution so that a first portion of the first feed solution enters a plurality' of pores in the shaped structure, allowing a mineral binder to form from the cementation reagent which binds at least two particles of the plurality of aggregate particles.

53. The system of claim 52, configured to be a continuously fed system, a batch system, or a fed batch system.

54. The system of claim 52, further comprising one or more of: a biologies activation system (3001); a mixing tank (102) (e.g.. a cement mixing tank); a staging tank (3005) (e.g., coupled to a system of pumps); an incubation system (106) (e.g., including one or more heat sources, sensors, humidifiers, fans, filters, etc.); a computing system (101); one or more pumps (3012); a conveyor system (120) (e.g., including a conveyor belt, a carousel, robotic arm conveyor, a monorail conveyor, an overhead conveyor, and / or a crane); a first cementation reagent source (e.g., a first cementation reagent tank, e.g., ; a second cementation reagent source; an acid source (e.g., an acid tank, e.g., configured to store hydrochloric or sulfuric acid, e.g., coupled to a system of pipes and pumps, e.g., connecting to the staging tank); a pH sensor; a concentration sensor (e.g., an EC sensor); a temperature sensor (e.g., a thermocouple, a thermistor, or a semiconductor-based sensor); a humidity' sensor;a heating and / or cooling system (e.g., including one or more fans, heating elements, cooling elements, etc.); a gas source (e.g., an oxygen source, a carbon dioxide source, etc., e.g., fluidly coupled to the biologies activation and / or expansion tank and / or the feed system); a clean-in-place system (e.g., including a tank configured to store one or more CIP reagents, a system of pumps and pipes to connect to, e.g., the staging tank, the feed system, an effluent system, etc.); an effluent system (3008) (e.g., including one or more filters (e.g., microfilters), a biologies inactivation system, an effluent tank, an effluent drain valve, etc.); or a regeneration system (3009) (e.g., including one or more permeable barriers, pumps, mixers, etc.).

55. The system of claim 52, further comprising one or more of: a feed bath (3101); an impulse delivery system (3103) (e.g., including a piston, an actuator, a plow, a diaphragm, a screw, or a blade, and / or a mechanism for positionally translating the shaped structure, e.g., one or more components of the conveyor system (e.g., the in-bath conveyor system), e.g., a conveyor belt, carouse, or crane); an in-bath conveyor system (3102); an inter-system conveyor system (3101) (e.g., including a finger car, a conveyor belt, and / or a robotic arm); an impulse control application (195) in the computing system (101); or a regeneration control application (196) in the computing system (101).

56. A construction material comprising: c) 70-95 wt% of an aggregate blend having an unpressed (e.g., tapped) green pycnometric intergranular voids percentage or porosity of less than 55%; and d) 5-30 wt% of one or more metal carbonates; wherein at least 5 wt% (e.g., 5-30 wt%) of the construction material is metal carbonates deposited by an enzymatic process after the aggregate blend has been formed into a shape.

57. The construction material of claim 56, wherein:the unpressed green pycnometric intergranular voids percentage or porosity of less than 50%. 45%. 40%. 35%. 33%. 34%. 32%. 31%. 30%. 20%, 15%, or less than 10% etc.

58. The construction material of claim 56, wherein: the aggregate blend is well-graded.

59. A construction material, comprising: c) 70-95 wt% of an aggregate blend that is a gap-graded aggregate blend; and d) 5-30 wt% of one or more metal carbonates; wherein at least 5 wt% (e.g., 5-30 wt%) of the construction material is metal carbonates deposited by an enzymatic process after the aggregate blend has been formed into a shape.

60. A construction material, comprising: an aggregate blend of two or more aggregates having a maximum particle size, D, a gradation curve on a plot of % of particles passing through a sieve of a sieve size vs the sieve size, and having an area thereunder that is ±20% of an area under a gradation curve of pi = (di / D)°45(Equation A), or pi = (di / D)°5(Equation B); wherein pi is a % of particles passing through an i* sieve size, di is the opening size of the ithsieve size; and5-30 wt% of one or more metal carbonates; wherein at least 5 wt% (e.g., 5-30 wt%) of the construction material is metal carbonates deposited by an enzymatic process within a matrix of the aggregate.

61. A construction material, comprising: an aggregate blend of two or more aggregates having a maximum particle size, D, a gradation curve on a plot of % of particles passing through a sieve of a sieve size vs the sieve size of pi = (di / D)°45(Equation A), or pi = (di / D)°5(Equation B); or within a least squares error of 0.00001 to 0. 1 of the gradation curve of EquationA or the gradation curve of Equation B; wherein pi is a % of particles passing through an i* sieve size, di is the opening size of the ithsieve size; and5-30 wt% of one or more metal carbonates; wherein at least 5 wt% (e.g., 5-30 wt%) of the construction material is metal carbonates deposited by an enzymatic process within a matrix of the aggregate.

62. A construction material, comprising: an aggregate blend of two or more aggregates in proportions that correspond within 10% to a maximum theoretical packing density for the aggregate blend; and5-30 wt% of one or more metal carbonates; wherein at least 5 wt% (e.g., 5-30 wt%) of the construction material is metal carbonates deposited by, or in conjunction with, an enzymatic process within a matrix of the aggregate.

63. The construction material of claim 62, wherein: the aggregate blend is a binary aggregate blend; or the aggregate blend is a ternary aggregate blend; or the aggregate blend is a quaternary aggregate blend.

64. The construction material of claim 62, wherein: a ratio of aggregates corresponds to a void content that is within 10% of a theoretical minimum void content for the three aggregates when blended and packed together.

65. The construction material of claim 62, wherein: a maximum theoretical packing density for the aggregate blend is that according to a continuous packing model or a discrete packing model (for example according to the Modified Andreasen- Andersen model, the Fumas model, or the Funk-Dinger model).

66. The construction material of claim 62, wherein: one or more aggregates of the aggregate blend is a moderate angularity aggregate or a high angularity aggregate (e.g., a roundness of <0.4 (e.g., 0. 1-0.4, 0.2-0.4, or 0.3-0.4), and / or a compactness of <0.8, and / or aspect ratio of >1.5: 1 (e.g., 1.5: 1 to 5: 1); or one or more aggregates of the aggregate blend is a moderate to high roundnessaggregate (e.g., from 0.4 to 0.6 roundness, e.g., about 0.5 roundness); or all aggregates of the aggregate blend are of high roundness (e.g., 0.5-1 roundness, e.g., about 0.6, 0.7, 0.8, 0.9, or 1 roundness) (for example an aggregate blend with a particle size distribution that produces an extremely high density / low inter-granular void volume, but comprised primarily of high roundness (e.g., spheroid) aggregate particles so that a high degree of pore interconnectivity remains, allowing feed to access pore network despite low aggregate pore volume).

67. The construction material of claim 62, wherein one or more aggregates of the aggregate blend has an uncompacted void % of from 34% to 50% (e.g.. 34-40%, 35-40%, 35-45%, 36-39%. 37-40%, 38-42%, 40-45%. 40-50%, 42-46%, 43-48%, or 45-50%).

68. The construction material of claim 62, further comprising: fibers (e.g., polymer fibers (e.g.. polycarbonate, polyvinyl chloride, nylon, polyester, polyethylene, polypropylene, polyacrylate, cross-linked polyvinyl alcohol, polyoxymethylene, natural fibers (e.g., jute, flax, hemp, banana, etc.), bioplastic fibers (e.g., polylactic acid, polyhydroxyalkanoates), mineral fibers (e.g., fiberglass, basalt fibers, etc.); hollow microspheres (e.g., polymer microspheres (e.g., plastic microspheres, hydrogel microspheres, resin microspheres), glass microspheres, ceramic microspheres (e.g., cenospheres); entrained gas (e.g., air); textiles (e.g., woven or non-woven networks of fibers of, e.g., jute, hemp, polypropylene, polyamides (e.g., polyaramids (e.g., para-aramids or meta-aramids) or polyphthalamides or aliphatic polyamides (e.g., nylon 6,6)); or a combination thereof.

69. The construction material of claim 62, wherein: the construction material retains a flexural strength of at least 80% of its original flexural strength after 25 cycles of an EN 14617 freeze / thaw test (e.g., has a KMF25 of at least50%, 60%, 70%, 75%, 80%, 85%, 90%, or 95%); and / or wherein the construction material has a flexural strength of at least 4.0 MPa after 25freeze-thaw cycles.

70. The construction material of claim 62, further comprising: 0.1-50 wt% of biochar.

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