Composition and method for manufacturing dust preventing material and construction material

Abstract

To provide a composition and a method for manufacturing a dust preventing material and a construction material containing high-embodied energy in place of concrete containing low-embodied energy.SOLUTION: There are provided a composition and a method for manufacturing a construction material and a dust preventing material using enzyme-producing cells, a certain amount of a nitrogen source such as urea, and a certain amount of calcium such as calcium chloride. Calcium contributes to the formation of calcium carbonate that forms a solid structure, a layer, or a shield. The composition according to the present invention can be blown to a surface of an object for controlling erosion thereof, supporting the foundation, preventing a sinkhole from being formed, or for other applications, or a different method can be applied in using the composition, Ammonia, water and other by-products of the process can be recycled or reused for the same purpose or for a different purpose including, for example, production of a fertilizer or an energy source, or can separately be produced using a selectively cultured microbe.SELECTED DRAWING: None

Description

[Technical field]

[0001] REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Application No. 62 / 200,288, filed August 3, 2015, U.S. Provisional Application No. 62 / 188,556, filed July 3, 2015, and U.S. Provisional Application No. 62 / 130,854, filed March 10, 2015, each of which is specifically incorporated by reference in its entirety.

[0002] 1. Field of the Invention The present invention relates to kits, compositions, tools and methods for building material production and for dust suppression. More specifically, the present invention relates to materials and methods for the production of bricks and masonry blocks and for dust suppression using isolated enzymes, enzyme-producing bacteria or cells, or spore producing enzyme-producing microorganisms. [Background technology]

[0003] 2. Background The built environment is primarily constructed using a limited set of traditional materials: clay, concrete, glass, steel, and wood. Masonry, widely used throughout history, still constitutes a large part of the built environment, both in load-bearing and veneer construction. According to Chaisson, traditional clay brick manufacturing industries collectively produce over 1.23 trillion units per year, which are highly dependent on non-renewable natural resources. Each unit of clay brick produced in coal kilns produces approximately 1.3 pounds of carbon dioxide. According to Burke, brick manufacturing collectively produces over 800 million tons of manmade CO2 annually, and this is just one of the materials currently used in building construction.

[0004] Fired clay bricks can be produced in 3 to 20 days, depending on the equipment and processes used. This range represents modern automated factories that can process bricks without manual labor, to the clamp method used in many developing countries, where bricks are piled around a roaring fire.

[0005] As an alternative to load-bearing fired clay stone, concrete masonry units [CMUs] are widely used because they are more economical, faster to manufacture, and can be used as a structural typology throughout a building. These units, made of concrete, are manufactured using Portland cement, coarse aggregate, and sand filler materials. According to Hanley of the U.S. Environmental Protection Agency, total carbon dioxide (CO2) emissions from cement production were approximately 829 million metric tons of CO2 in 2000.

[0006] These traditional materials contain high embodied energy, and the components of concrete and steel are mined from non-renewable sources. Approximately 40 percent of total carbon dioxide is associated with the building industry, primarily due to the production and disposal of materials. Biologically grown materials, when produced as part of a local ecosystem, can be pollutant-free and contain low embodied energy.

[0007] Natural cements are produced through chemical processes associated with chemical deposition and weathering and can be found in various locations in the Earth's crust. The formation of natural sandstone is primarily due to the precipitation of calcite cement. As an alternative to natural deposition, the formation of natural cement has been carried out using urease-producing Sporosarcina Pasteurii, a non-pathogenic common soil bacterium that has the ability to induce the production of calcite through chemical reactions. The result is a hardened material that forms in a process termed microbially induced calcite precipitation [MICP] by Stocks-Fischer. Applications include environmental stabilization of contaminated soils and encapsulation of hazardous materials and other contaminants in natural soils and acid mine tailings. Ramachandran and Jonkers proposed the use of microorganisms to repair cracks in concrete structures and for the repair of cracks in monuments. According to DeJong and Whiffin, civil engineering researchers in the United States, Australia and the Netherlands have advocated the use of MICP for soil stabilization and erosion control.

[0008] There is a need for a process for producing building materials that uses readily available materials and is economical and environmentally safe, without the high energy consumption associated with the production of clay bricks and other traditional stone substitutes. Summary of the Invention

[0009] The present invention overcomes the problems and shortcomings associated with current strategies and designs and provides new tools, compositions and methods for the production of building materials.

[0010] One embodiment of the present invention relates to a composition comprising a support material to which urease producing cells or urease producing cell spores are bound and a transport medium and optionally a nutrient mixture. Preferably, the support material is organic or inorganic and includes rock, glass (e.g., Poraver), wood, paper, metal, plastic, polymer, mineral or combinations thereof. Preferably, the composition is a liquid, gel, sludge, pumpable slurry, dry powder or crystals and the support material is in the form of beads, grains, rods, strands, fibers, flakes, crushed or crushed stone, crystals, granules or combinations thereof. Preferably, the support material is sand, glass, wood (e.g., residues, pulp, sawdust, lignin), metal, polymer, fines (e.g., microcellulose), waste (e.g., ash, scrubber waste, residues), co-cultured microorganisms or combinations thereof, and the urease producing cells or urease producing cell spores include yeast, algae, bacterial or eukaryotic cells, cell spores, anaerobic cells or facultative anaerobic cells. Preferred bacteria are Sporosarcina pasteurii, Sporosarcina ureae, Proteus vulgaris, Bacillus sphaericus, Myxococcus xanthus, Proteus mirabilis, Helicobacter pylori, or variants, serotypes, mutants or combinations thereof, and preferred yeast, algae, bacteria or eukaryotic cells or cell spores are genetically engineered. The support and cells are preferably bound by hydrophobic, hydrophilic, ionic, non-ionic, covalent bonds, van der Waals forces or combinations thereof, and / or the support is at least partially or completely covered by a film that promotes binding of the urease producing cells. Preferred films include a polymer or cell nutrients, and preferably the composition includes a colorant which may be red, blue, green, yellow or any combination or shade thereof.Preferably, the composition comprises an identifying agent or detectable marker, such as a microscopic tag, a color, a nucleic acid or peptide, an enzyme or another substance.

[0011] Another aspect of the present invention relates to a kit for producing solid moldings, the kit comprising the composition of the present invention; a second composition comprising nutrients for the growth of urease producing cells and / or the germination of cell spores; a plurality of formwork sets, each set encircling the shape of at least one solid molding and comprising one or more porous panels; and a third composition comprising a calcium source (e.g., CaCl2), a nitrogen source (e.g., urea) or both a calcium source and a nitrogen source. Preferably, the kit is for making solid moldings, such as rectangular, square, circular, oval or irregular shapes. Preferred solid moldings include, but are not limited to, blocks, boards, bricks, paving stones, panels, tiles or veneers. Preferably, the kit of the present invention is for producing blocks, such as concrete masonry blocks, cinder blocks, foundation blocks, breeze blocks, hollow blocks, solid blocks, besser blocks, clinker blocks, high or low density blocks, or cellular blocks. Preferably, the nutrients comprise amino acids, proteins, polysaccharides, fatty acids, vitamins and minerals.

[0012] Another aspect of the present invention relates to a method for producing a solid molding, the method comprising the steps of: mixing the composition of the present invention with aggregate material and water to form a mixture, the aggregate material being composed mainly of particles with a diameter of 5 mm or more or particles with a diameter of 5 mm or less (e.g., granules); optionally distributing the mixture into a plurality of moulds, each mould comprising at least one porous panel; adding a second composition to the mixture, the second composition comprising nutrients that promote the growth of urease-producing cells; adding a third composition to the mixture, the third composition being a liquid comprising calcium; incubating the mixture for a period of time to allow covalent bonds to form between the particles; and removing the solid molding from the mould. Preferably, the aggregate material comprises rock, glass, wood, paper, metal, plastic, polymer, mineral or a combination thereof, and / or the mixing step comprises spraying the composition as a liquid onto the aggregate material. Preferably, the mould is substantially submerged during the incubating step, and air is bubbled into the submerged mould. Preferably, the third composition is repeatedly added to the mixture during the incubating step, which is drained through the bottom panel and optionally recycled. Preferably, the incubating step is carried out under ambient conditions, and the third composition comprises calcium chloride, calcium acetate, calcium phosphate, calcium carbonate, calcium lactate, calcium nitrate or a calcium salt. Preferably, the pH of the mixture is monitored during the incubating step. Preferably, the solid molding is a block, board, brick, thin brick, paving stone, panel, tile or veneer, stone (artificial stone, cultured stone, colored stone), and the mixture further comprises fibers or nanofibers, e.g. fibers or nanofibers of wood, glass, plastic, metal or polymer. Preferred fibers include, e.g., polypropylene, HDPE, carbon fibers, including high strength carbon fibers, rayon, and biodegradable and non-biodegradable fibers, e.g., polymers such as polylactic acid, cellulose, minerals, chitin, lignin and other plant materials. Preferably, additional nutrients are added during the incubating step, and the solid molding removed from the formwork is dried.

[0013] Another aspect of the invention includes a composition comprising urease producing cells or urease producing cell spores encapsulated or coated with a nutrient medium, such as a protein or polysaccharide, or a polymer, such as polylactic acid, which is water soluble. Preferably, the nutrient medium further comprises additional urease producing cells or urease producing cell spores.

[0014] The basic features and various aspects of the present invention are listed below. [1] A composition comprising a support material to which urease-producing cells or urease-producing cell spores have been applied and a transport medium. [2] The composition of [1], wherein the support material comprises organic or inorganic matter, rock, glass, wood, paper, metal, plastic, polymer, fiber, mineral, or combinations thereof. [3] The composition of [1], which is a liquid or a dry powder. [4] The composition of [1], wherein the support material is in the form of beads, grains, rods, strands, fibers, flakes, fibres, crushed or crushed stone, crystals or combinations thereof. [5] The composition of [1], wherein the support material is sand, glass, fiberglass, paper, ash, wood, metal, waste or a combination thereof. [6] The composition of [1], wherein the urease-producing cells or urease-producing cell spores comprise yeast, algae, bacteria or eukaryotic cells or cell spores. [7] The composition of claim 6, wherein the bacterium is Sporosarcina pasteurii, Sporosarcina ureae, Proteus vulgaris, Bacillus sphaericus, Myxococcus xanthus, Proteus mirabilis, Helicobacter pylori, or a variant, serotype, mutant or combination thereof. [8] The composition of claim 6, wherein the yeast, algae, bacteria or eukaryotic cells or cell spores are genetically engineered. [9] The composition of [1], wherein the support material and the cells are bound by hydrophobic bonds, hydrophilic bonds, ionic bonds, non-ionic bonds, covalent bonds, van der Waals forces, or combinations thereof.

[10] The composition of [1], wherein the support is at least partially covered by a film that promotes attachment of urease-producing cells.

[11] The composition of

[10] , wherein the film comprises a polymer or cell nutrients.

[12] The composition of [1], further comprising a colorant.

[13] The composition of [1], comprising an identifying agent or detectable marker.

[14] The composition of [1], wherein the transport medium comprises components for cell growth.

[15] The composition of

[14] , wherein the components for cell growth include one or more of water, nutrients, vitamins, minerals, amino acids, proteins, oils, fatty acids, sugars and polysaccharides.

[16] A kit for producing a solid molded product, comprising: [1] The composition of a second composition comprising nutrients for the growth of urease producing cells and / or the germination of cell spores, a plurality of form sets, each set enclosing at least one solid molding shape and including one or more porous panels; and a third composition comprising calcium, nitrogen, or both calcium and urea; Including the kit.

[17] The kit of

[16] , wherein the solid molding is rectangular, square, circular, elliptical or irregularly shaped.

[18] The kit of

[16] , wherein the solid molding is a block, board, brick, paving stone, panel, tile or veneer.

[19] The kit of

[18] , wherein the blocks are concrete masonry blocks, cinder blocks, foundation blocks, breeze blocks, hollow blocks, solid blocks, besser blocks, clinker blocks, high or low density blocks, or cellular blocks.

[20]

[16] kit, whose nutrients include amino acids, proteins, polysaccharides, fatty acids, vitamins and minerals. [twenty one] A method for producing a solid molded product, comprising the steps of: mixing the composition of [1] with aggregate material and water to form a mixture, the aggregate material being composed primarily of particles less than 50 mm in diameter; distributing the mixture into a plurality of forms, each form including at least one porous panel; adding a second composition to the mixture, the second composition comprising nutrients that promote the growth of urease producing cells; adding a third composition to the mixture, the third composition being a liquid comprising calcium; incubating the mixture for a period of time to allow covalent bonds to form between the particles; and Removing the solid molding from the mold A method comprising: [twenty two] The method of

[21] , wherein the aggregate material comprises rock, glass, wood, paper, metal, plastic, polymer, mineral or combinations thereof. [twenty three] The method of

[21] , wherein the mixing step includes spraying the composition as a liquid onto the aggregate material. [twenty four] The method of

[21] , wherein the mold is substantially immersed during the incubating step. [twenty five]

[24] , in which air is bubbled into a submerged formwork.

[26] The method of

[21] , wherein additional third composition is repeatedly added to the mixture during the incubating step, drained through a bottom panel, and optionally recycled.

[27] The method of

[21] , wherein the incubating step is carried out under ambient conditions.

[28] The method of

[21] , wherein the incubating step is carried out at a temperature between 5°C and 50°C.

[29] The method of

[21] , wherein the third composition comprises calcium chloride, calcium acetate, calcium phosphate, calcium carbonate, calcium lactate, calcium nitrate or a calcium salt.

[30] The method of

[21] , wherein the pH of the mixture is monitored during the incubating step.

[31] The method of

[21] , wherein the solid molding is a block, board, brick, paving stone, panel, tile or veneer.

[32] The method of

[21] , wherein the mixture further comprises fibers or nanofibers.

[33] The method of

[32] , wherein the fibers or nanofibers are composed of wood, glass, plastic, metal or polymer.

[34] The method of

[21] , wherein additional nutrients are added during the incubation step.

[35] The method of

[21] , wherein the solid molding removed from the mold is dried.

[36] A composition comprising urease producing cells or urease producing cell spores encapsulated, microencapsulated or coated with a nutrient, protein, polysaccharide, polymer or other medium.

[37] The composition of

[36] , wherein the nutrient medium further comprises additional urease-producing cells or urease-producing cell spores.

[38] A composition comprising urease and / or urease-producing cells, a nitrogen source, a calcium source and water.

[39] The composition of

[38] , further comprising an aggregate material having an average diameter of less than 0.05 mm.

[40] The composition of

[38] , further comprising an aggregate material having an average diameter of 1 nm to 40 nm.

[41] The composition of

[38] , further comprising an aggregate material having an average diameter of 1.0 mm to 50 mm.

[42] The composition of

[38] , wherein the nitrogen source is urea and the calcium source is calcium chloride.

[43] A composition according to

[38] , comprising a quantity of urease and / or urease producing cells, a quantity of a nitrogen source, a quantity of a calcium source and a quantity of water, which provides for solidification of the composition within a set time.

[44] The composition of

[38] comprising urease-producing cells and further comprising nutrients that promote cell growth.

[45] 11. A method for layering a solid object with calcium carbonate, comprising contacting the solid object with the composition of

[38] and promoting the formation of calcium carbonate.

[46] The method of

[45] , wherein the contacting step comprises spraying the composition onto a solid object and / or maintaining a desired vapor pressure.

[47] The method of

[45] , wherein the contacting step comprises immersing the solid object in the composition, spraying the solid object with the composition, misting the solid object with the composition, or exposing the solid object to steam along with the composition.

[48] A dust prevention method comprising the step of spraying the composition of

[38] onto a surface.

[49]

[48] ​​method, in which the blasted surface is more resistant to erosion compared to the non-blasted surface.

[50] The method of

[48] , wherein the surface is a sidewalk, pile, cliff or roadway. Other aspects and advantages of the invention will be set forth in part in the description that follows, and in part will be obvious from the description, or may be learned by practice of the invention. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0015] Description of the invention Traditional building materials, such as clay bricks and concrete, require huge amounts of energy during the manufacturing process. These processes are heavily dependent on the burning of natural resources, such as oil, coal, and wood. This dependency consumes huge amounts of energy sources and simultaneously generates large amounts of carbon dioxide, thus relying heavily on limited energy sources. Alternative methods that require much less energy during production have been reported, utilizing enzymes produced by microbial cells. Typically, the cells are aerobic and / or facultative anaerobic cells, including, for example, Sporosarcina pasteurii, Sporosarcina urea, Proteus vulgaris, Bacillus sphaericus, Myxococcus xanthus, Proteus mirabilis, Helicobacter pylori and other strains, serotypes, variants, mutants, and CRISPR modifiers (clustered regularly interspaced short palindromic repeats). The cells produce urease, an enzyme that forms calcite crystals in the presence of calcium and nitrogen sources. This process is commonly referred to as microbially induced calcite precipitation (MICP) and can be carried out using cells or purified enzymes. It requires little or no heating, making it energy-saving in both cost and efficiency.

[0016] Enzymes and / or enzyme producing cells that catalyze the production of ammonia and carbon dioxide and raise the pH level of the composition are dispersed in a composition that includes a nitrogen source and a calcium source, such as urea and calcium chloride, along with aggregate material. The increase in pH forms an inorganic precipitate with combined calcium and carbon dioxide. The cells or other particles act as nucleation sites and attract inorganic ions from the calcium to the particle surface to form calcium carbonate crystals, such as calcite crystals or other calcium carbonate polymorphs. This mineral growth fills the interstices between the aggregate particles and biocements or bonds the aggregate particles together to form a solid mass. The resulting material exhibits similar composition and physical properties to naturally formed sandstone, but its hardness can be predetermined based on at least the structure of the initial components and the desired pore size.

[0017] Surprisingly, new tools, compositions, techniques and methods have been discovered for the production of solid objects formed from aggregate materials using the MICP process. Surprisingly, it has been discovered that compositions can be made that include a support material to which urease producing cells or urease producing cell spores are attached, which may optionally include a nutrient composition. Contacting the cells or enzyme with the support material allows for commercial exploitation of the MICP process for mass production and manufacturing of building materials. Commercial sources of urease include, for example, jack beans. The enzyme may be maintained as a liquid, but is preferably lyophilized for ease of storage and transport, and rehydrated with water, buffered water or another hydrating agent that maintains enzyme activity prior to use. Preferably, the pure enzyme is encapsulated in microshells or spheres of sugars, lipids or other polymers. Encapsulation techniques include, for example, encapsulation with nano-organized microshells and encapsulation with xanthan-alginate spheres. The preferred enzyme concentration is 0.5-5 mg / ml in 0.1 M phosphate buffer at pH 7.6. Preferred enzyme concentrations are about 0.1-100 mg / ml, more preferably about 0.5-3.0 mg / ml, more preferably about 0.5-2.0 mg / ml and more preferably about 1.0 mg / ml. The enzyme may be further diluted to obtain a rate of 0.02-0.04 ΔA / min prior to use. Enzyme activity is determined by the following reaction, which relates ammonia production to the glutamate dehydrogenase reaction: Thus, one unit of enzyme will result in the oxidation of 1 micromole of NADH per minute at 25° C. and pH 7.6.

[0018] This method of producing building materials through the induction of cementation exhibits low embodied energy and can be carried out under ambient or higher or lower pressures and ambient or higher or lower temperatures. For example, preferred pressures are from about 10 psi to about 100 psi and all pressure values ​​therebetween, and preferably from about 14 psi to about 50 psi. Higher pressures can also be utilized, but are not usually required for the required energy expenditure. Preferred temperature ranges are at least -20°C to greater than 80°C, preferably from about 5°C to about 50°C, preferably from about 15°C to about 30°C, preferably from about 20°C to about 25°C. Preferably, the temperature range is below 30°C, below 40°C, below 50°C, below 60°C, or below 70°C. The ambient temperature and conditions as well as the available aggregate content may determine whether pure enzymes, lyophilized enzymes, spores, or live cells can be utilized as starting components. Live cells may be used under temperatures where mild weather conditions exist, while pure enzymes may be advantageous under more extreme conditions of low or high temperatures. Spores are used where immediate mineralization is not required and the spores are given sufficient time to germinate and express the enzyme.

[0019] The process also involves the production of significant amounts of by-products, such as ammonia, but not all of these are utilized in the calcite formation. The inclusion of a wastewater recovery system in conjunction with this manufacturing method is another aspect of the present invention. Recovery of ammonia from the wastewater converts it into ordinary water that can be reused or disposed of without the need for further decontamination procedures. Preferred ammonia recovery methods include, for example, ion exchange resins and commercially available processes such as ammonia electrolysis, zeolites, clinoptilolite, and combinations thereof. Preferably, the recovered ammonia can be utilized as a fertilizer, converted to nitrogen, utilized for energy production, or utilized in other applications.

[0020] One embodiment of the present invention relates to a composition comprising urease producing cells or urease producing cell spores, urease enzyme (e.g. crude extract or unpurified or purified enzyme) in a transport medium and optionally a nutrient medium. The transport medium includes, for example, a growth medium for the urease producing cells and / or other support cells, an enzyme stabilization medium, a reagent medium, a buffered solution and combinations thereof. The composition may include or be combined with a support material, which may be organic or inorganic, preferably solid or semi-solid, preferably containing holes or perforations and / or otherwise porous. Organic support materials include, for example, biomass, such as, preferably, moss, hay, straw, grass, branches, leaves, algae, dirt, ash, dust, granular matter, litter and combinations thereof. Inorganic support materials include, for example, minerals, supplemental cementitious materials (SCM), crushed or crushed rock, fines and combinations thereof. Fibrous materials include sheets or tarps of burlap, paper, wood (e.g., residues), cotton or other natural or synthetic fibers. Non-natural and man-made materials, such as sheets of plastic, glass, fiberglass, vinyl, rubber, synthetic fibers or combinations thereof, may also be used. To this solid support, urease producing cells, urease enzyme or simply other cells are applied or otherwise introduced. Preferably, these other cells are useful to support the growth of the urease producing cells or to enhance chemical processes involved in and not otherwise interfere with the MICP process, or to act as nucleation sites. Preferably, these other cells are natural or latent microorganisms in the local environment or provided with the mixture, are non-pathogenic, non-toxic and / or relatively harmless in the amounts used, and are readily available, present in, or provided in the local environment. Cells can be grown directly on the support for the production of building tools and products, and at the desired density or stage of growth, the organic material is uniformly distributed and / or thoroughly mixed into the aggregate material.Inorganic materials that may be used include, for example, rock (e.g., granules), sand, glass, wood, paper, metal, plastic, polymer, minerals, manufacturing or processing waste such as ash, carbon, or wood residues, any of which may be crushed or used in whole or in combination. Compositions may also be formed from waste that would otherwise be hazardous (e.g., radioactive materials, materials containing hazardous metals or toxic content, contaminated materials from scrubbers, or other hazardous materials) and formed into solid structures that can be stably stored or otherwise safely disposed of.

[0021] The compositions of the invention may be sprayed or otherwise applied to sheets or mats or natural or non-natural materials, and may be sprayed or otherwise applied to sheets used to prevent erosion, for example by forming a calcium carbonate crust on surfaces, piles, cliffs or other structures exposed to erosion. Using perforated or porous sheets or mats, the crust is formed over the support, adhering the material on which the sheet is placed. Nucleation sites for calcite formation may include, for example, polymers, fibers, granules, SCM, added Portland cement, powders, co-cultured microorganisms and combinations thereof. One or multiple layers of crusts may be formed at the site. In this case, erosion and dust prevention may be substantially reduced or eliminated in a defined area. Importantly, in this manner, the sheets may be easily replaced over time and / or fresh compositions of the invention may be reapplied to the surface as needed or desired. The mats provide the added benefit of "seeding" the site for rehabilitation after work is completed, thereby allowing restoration work to proceed at the site, for example, by returning the site to its natural state. This applies in particular to mining sites where mining operations have ceased.

[0022] In a preferred embodiment of the invention, the composition of the invention is applied to the surface area as a liquid, gel, slurry, sludge, semi-solid or dry powder. The spores and / or microorganisms of the composition of the invention produce enzymes that catalyze the formation of calcium carbonate exoskeleton in the presence of liquid, preferably water, buffered water or another aqueous substance. A nutrient mixture appropriate for the individual microorganism may be included with the cells. When the composition dries, the exoskeleton remains and the cells become dormant. If sufficient nutrient and / or substrate material is present, the cells will self-propagate and form a new exoskeleton whenever sufficient aqueous liquid is provided. In a preferred embodiment, the nutrient and / or substrate material may be dispersed on the surface area of ​​interest in a time- or sustained-release form, for example as a dry component with a predetermined dissolution rate. Reformation may simply be a matter of reapplication of water that dissolves the nutrient and / or substrate, thereby reactivating the microorganism. The reactivated microorganism produces enzymes that form the exoskeleton. This process may be repeated with or without reapplication of the microorganism, nutrient and / or substrate, or with only occasional additions. This process can be linked to meteorological phenomena such that rain provides a water source. By providing microorganisms and / or spores and providing a composition that includes slow-release nutrients and / or substrates, the exoskeleton can be reformed repeatedly in an area over an extended period of time.

[0023] Preferably, the composition of the invention, including all necessary components, such as microorganisms, spores and / or enzymes, nitrogen source, and calcium source, and optionally nucleation sites (e.g., powders, granules, co-cultured microorganisms and / or other materials), is applied to a surface, such as an unpaved road, or a structure, such as a hill or cliff. The microorganisms grow and produce enzymes that catalyze the formation of calcium carbonate crusts on the road surface. As vehicles pass over the road, the crust breaks down, and eventually the crust becomes dust. Periodically, the geographic area experiences rain or other forms of precipitation that dissolve the slow-release nutrient and / or substrate material, thereby promoting the growth of dormant microorganisms. The microorganisms produce enzymes that catalyze the formation of new crusts on the road. During periods of low precipitation, an aqueous solution, which may or may not contain additional nutrient and / or substrate material, is reapplied to the road to activate the microorganisms. Preferably, the composition is a liquid, gel, slurry, sludge or dry powder, and the support material can be in the form of beads, grains, rods, strands, fibers, flakes, dirt, biomass, sand, crushed or crushed stone, fines, supplementary cementitious materials (SCM), crystals, co-cultured microorganisms, or combinations thereof. The size of the fines is preferably 250 mesh or less, more preferably 200 mesh or less, more preferably 150 mesh or less, or more preferably 100 mesh or less (reference examples include beach sand mesh size=700, fine sand mesh size=250; Portland cement mesh size=74; silt mesh size=44; soot mesh size=2). The support material and aggregate material can be the same or different. Preferably, the support or aggregate material is sand, glass, metal, added Portland cement, SCM, granules, co-cultured microorganisms (e.g., natural, latent, local, added or genetically engineered microorganisms) or combinations thereof, and the urease-producing cells or urease-producing cell spores include yeast, algae, anaerobic cells, facultative anaerobic cells, bacterial or eukaryotic cells or cell spores.Preferred bacteria are Sporosarcina pasteurii, Sporosarcina urea, Proteus vulgaris, Bacillus sphaericus, Myxococcus xanthus, Proteus mirabilis, Helicobacter pylori, or variants, serotypes, mutants or combinations thereof, and preferred yeast, algae, bacteria or eukaryotic cells or cell spores are genetically engineered. Other enzyme-producing bacteria capable of biocementation include Sporosarcina urea, Proteus vulgaris, Bacillus sphaericus, Myxococcus xanthus, Proteus mirabilis and Helicobacter pylori, although appropriate caution should be exercised for pathogenic strains. Combinations of any of these strains as well as functional variants, mutants and genetically engineered strains may also be used. The support material and cells are preferably bound by hydrophobic, hydrophilic, ionic, non-ionic, covalent bonds, van der Waals forces or a combination thereof, and / or the support material is at least partially or completely covered by a film that promotes the binding of the urease producing cells. Preferred films include polymers or cell nutrients, and preferably the composition includes a colorant, which may be red, blue, green, yellow or any combination or dark color thereof. Preferably the composition includes an identifying agent or detectable marker, such as a microscopic tag, a color, an enzyme or another substance.

[0024] The support and / or aggregate materials may contain additional components that provide benefits to the building material. For example, they may contain enzymes, cofactors and / or other chemicals that generate useful and / or additional nucleation sites and / or additional cells (e.g., naturally occurring, local or latent bacteria, yeasts, eukaryotic cells, algae and recombinant variants thereof) that break down stains that occur in and / or on the final product. Stains include those from air pollution, soot, mold or animal waste. Alternatively, the chemicals or enzymes may impart color, texture or a desired function or appearance to the final product.

[0025] Another aspect of the present invention relates to a kit for producing solid moldings, the kit comprising the composition of the present invention, a second composition comprising nutrients for the growth of urease producing cells and / or the germination of cell spores; a plurality of formwork sets, each set encircling the shape of at least one solid molding and comprising one or more porous panels; and a third composition comprising a calcium source (e.g., CaCl2), a nitrogen source (e.g., urea) or both a calcium source and a nitrogen source. Preferably, the kit is for making solid moldings, such as rectangular, square, circular, oval or irregular shapes. Preferred solid moldings include, but are not limited to, blocks, boards, bricks, paving stones, panels, tiles, countertops or veneers. Preferably, the kit of the present invention is for producing blocks, such as concrete masonry blocks, cinder blocks, foundation blocks, breeze blocks, hollow blocks, solid blocks, besser blocks, clinker blocks, high or low density blocks, or cellular blocks, thin bricks, artificial stones, cultured stones or colored stones. The nutritional composition of the present invention may comprise a nutrient medium that maintains cells and / or allows cells to grow and proliferate well.Various types of nutrient media for the cells of the present invention, particularly bacterial cells, are known and commercially available, including at least the minimal medium (or transport medium) typically used for transport to maintain vitality without propagation, and yeast extract, molasses and corn steep liquor typically used for growth and propagation.Preferably, the nutrients comprise amino acids, proteins, polysaccharides, fatty acids, vitamins and minerals.

[0026] Another aspect of the present invention relates to a method for producing a solid molding, the method comprising the steps of: mixing a composition of the present invention with an aggregate material and water to form a mixture, the aggregate material being composed primarily of particles 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); distributing the mixture into a plurality of forms, each form comprising at least one porous panel; adding a second composition to the mixture, the second composition comprising nutrients that promote the growth of urease producing cells; adding a third composition to the mixture, the third composition being a liquid, powder or paste comprising calcium; incubating the mixture for a period of time to allow covalent bonds to form between the particles; and removing the solid molding from the form. Alternatively, the compositions may be combined and added en masse to the material in the form or combined with the material prior to addition to the form.

[0027] Another aspect of the invention relates to the structure and composition of the mold. The preferred mold comprises a thermoplastic material that can be molded or extruded into the desired shape. Preferred thermoplastics include, but are not limited to, plastics such as polypropylene, polystyrene, polyethylene, including HDPE (high density polyethylene), LPDE and recycled LDPE (low density polyethylene) and cross-linked polyethylene, glass and virtually any moldable polymer. Preferably, the polymeric material is provided as pellets or lenticular shapes of various thicknesses and uniformity. The pellets are filled into a porous mold and exposed to steam under pressure (the mold is not pressurized, pressure comes only from the steam). The resulting product provides material with a specified flow direction, and changes in gradient affect the flow direction, velocity and residual saturation.

[0028] Another aspect of the invention relates to compositions and structures that do not require a form (e.g., frameless manufacturing), where the structures are formed from a combination of the components of the invention and polymers and / or thermoplastics that are compacted by a compaction device and maintain the desired structure. Preferred compaction devices include hydraulic presses, with preferred pressures of 100 psi or more, 250 psi or more, 500 psi or more, 1000 psi or more, 2000 psi or more, 3000 psi or more, 4000 psi or more, 5000 psi or more. Preferred components of the invention include all components that form calcium carbonate structures in the form of a sludge or paste. The compaction device compacts these components under applied pressure into a molded object that is maintained and dried without significant change in the resulting shape. Preferred polymers and thermoplastics include, but are not limited to, plastics such as polypropylene, polystyrene, polyethylene including HDPE (high density polyethylene), LPDE and recycled LDPE (low density polyethylene) and cross-linked polyethylene, glass, sugars such as starch, lignin and almost any moldable polymer. Compacted moldings can be readily made from dilute slurries or sludges and maintain their shape during calcite formation. Preferably, calcite formation is accomplished in a steam chamber (e.g., at pressures higher than ambient pressure) with high steam pressure or sprayed or misted, the steam, mist or spray preferably containing nutrients or chemical substrates. Preferred moldings include blocks, bricks, thin bricks, artificial or cultured stones, paving stones or any useful structure.

[0029] Preferably, the multiple forms or compaction devices produce 5, 10, 50, 100, 500, 1,000, 10,000, 100,000, 1,000,000 or more moldings at a time. The number of forms or compaction devices that can be utilized simultaneously is limited only by the complexity of the equipment and the available space. These forms or devices can be stacked or provided in a single layer or on a pallet. The forms can have vertical walls that are connected together to form a space between them to receive the aggregate material. The forms can also have a floor, or the bottom of the form can be open if supported by a porous surface, such as soil, or the aggregate and composition can be mixed and pressed or extruded in a mold. Preferably, the vertical walls, at least the inner surface thereof, are made of a non-reactive, non-porous material or coating, such as cast or extruded acrylic resin. This allows the building material or brick to be easily removed from the form after it has hardened. Additionally, the vertical walls and floor of the form or pressing apparatus may have designs that create a surface texture (eg, lines, circles, waves, grooves, sketches, images, etc.) in the resulting brick or other structure.

[0030] Preferably, the aggregate material comprises rock, glass, glass fibre, wood (e.g. residues, pulp, sawdust, lignin), biomass, paper, metal, plastic, polymer, rubber, synthetic rubber, vinyl, mineral, co-cultured microorganisms, waste (e.g. ash, carbon, scrubber waste, radioactive pellets) or a combination thereof, and / or the mixing step comprises spraying the composition as a liquid onto the aggregate material. Preferably, the formwork is substantially submerged during the incubating step, and air is bubbled into the submerged formwork. Preferably, a third composition is repeatedly added to the mixture during the incubating step, which is drained through the bottom panel and optionally recycled. Preferably, the incubating step is carried out under ambient conditions, and the third composition comprises calcium chloride, calcium acetate, calcium phosphate, calcium carbonate, calcium lactate, calcium nitrate or a calcium salt. Preferably, the pH of the mixture is monitored during the incubating step. Preferably, the solid molding is a block, board, brick, paving stone, panel, tile or veneer, and the mixture further comprises fibers or nanofibers, e.g., wood, glass, plastic, metal or polymer fibers or nanofibers. The solid molding may be partially or uniformly porous, comprising a network of holes or gaps. The holes may be of a predefined size and / or structure, e.g., at least 5 microns, at least 10 microns, at least 20 microns or at least 50 microns in diameter, etc. Alternatively, the solid molding may be manufactured using a material that provides substantially no or only few holes. For example, the addition of a non-porous material to the aggregate mixture may generate complex and extended pathways that render the molding fluid impermeable.

[0031] Another aspect of the invention includes a composition comprising urease producing cells or urease producing cell spores coated with a nutrient medium. Preferably, the nutrient medium further comprises nutrients to promote the growth of additional urease producing cells or urease producing cell spores, and / or additional cells added to the aggregate that are beneficial to the end product.

[0032] Another aspect of the present invention relates to compositions, methods and systems for the treatment of aggregate material composed of particles with a composition comprising one or more of a nitrogen source, such as urea, a calcium source (e.g., calcium ions), and urease or urease-producing cells. Preferably, the particles have a diameter (e.g., actual diameter, average diameter 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 one preferred embodiment, the aggregate material may also be about 1 mm or less, preferably about 0.5 mm or less, more preferably about 0.1 mm or less, more preferably about 50 μm or less. Particularly preferred particle sizes include about 10 μm to about 1 mm, about 100 μm to about 0.5 mm, about 200 μm to about 1 mm, about 1 μm to about 200 μm, about 10 mm to about 1 μm, and about 10 nm to about 40 nm, as well as various combinations thereof. The particles include, for example, spores, coal dust, dust or soot from cement or brick manufacturing, cement block manufacturing, foundry operations, limestone grinding, sand beneficiation, mining, smelting, dye manufacturing, and other manufacturing process by-products, such as slag. The particles can be obtained and recovered from available or implemented dust control procedures. The particles can be mixed sizes, including, but not limited to, sizes above the preferred size, sizes below the preferred size, and combinations and mixtures of the preferred sizes. The particles that are aggregates and larger particles can include recycled and / or recyclable materials. The nitrogen source of the composition can be a single chemical, such as urea of ​​any grade and purity, preferably commercially available. Calcium ions are preferably obtained from commercially available sources, such as calcium chloride, etc. Urease enzyme or urease-producing bacteria may be included in the composition.Urease producing bacteria include, but are not limited to, the bacteria Sporosarcina pasteurii, Sporosarcina urea, Proteus vulgaris, Bacillus sphaericus, Myxococcus xanthus, Proteus mirabilis, Helicobacter pylori and combinations thereof. Urease producing cells include non-viable cells containing the enzyme, such as micelles, cells composed of lipids or fatty acids, and cells containing urease. Urease and / or urease producing cells may produce or release a predefined amount of enzyme for a defined period of time. Preferably, the amount of urease released per cell is fast enough to achieve rapid production of calcium carbonate in the presence of nitrogen and calcium ions.

[0033] Preferably, the particles are combined with a nitrogen source (e.g., urea), urease and / or urease producing cells, calcium ions, and preferably water to form a homogeneous slurry. The slurry can be painted or sprayed onto objects and / or surfaces, thereby forming a layer or shell, or molded into a molding that solidifies into an object that may be fully or partially solid, or pooled for dipping or submersion of the object to be coated with the slurry material, which also forms a layer or shell on the object surface. The object may include one or more layers, as desired, which may be permeable or impermeable to water or may improve resistance to weather conditions, such as sun damage, abrasion from snow, ice, and rain. Slurries that provide improved resistance are preferably constructed with aggregate materials, particles less than 0.1 mm in diameter. When the liquid dries, calcium carbonate bonds are formed between the particles and / or between the particles and the object. As a result, an object or molded structure can be obtained that includes a hardened calcium carbonate shell. Objects that may be produced in accordance with the present invention and / or layered with a shell or coating of the present invention include, but are not limited to, bricks, cement blocks, paving stones, countertops, glass, fiberglass, polymeric and acrylic structures, siding, walls, garden ornaments, slate and rock structures, tiles, paving stones, stairs, roofing, gutters, cement walls and planks, patios, balconies, fences and combinations thereof.

[0034] Another preferred embodiment of the present invention involves the production of ammonia and / or other compounds (e.g., ammonia, organic acids, alcohols, phenols, sulfides) by fermentation of microorganisms (e.g., microorganisms producing ammonia monooxygenase, hydroxylamine oxidoreductase, nitrifying bacteria). Preferably, the microorganisms are selectively cultured to maximize production of the desired enzymes. Hyper ammonia-producing microorganisms include, for example, ruminant-derived microorganisms, enteric microorganisms, Peptostreptococcus species, Clostridium species, Calliandra species, Atopobium species, Desulfomonas species, etc. The isolated ammonia can be reused or utilized in other processes, for example, in fertilizer and energy production.

[0035] Another aspect of the present invention includes spraying the slurry of the present invention onto natural or artificial geological surfaces, such as cliffs, sand dunes, aggregate piles, rock shelves, buttresses, ores, foundations, mining products, tailings, waste piles from manufacturing processes, or other structures where additional support or structuring is desired. Such support is advantageous in terms of convenience and economy compared to providing additional support for the structure of interest by conventional building systems. Additionally and preferably, the slurry of the present invention can be applied to geological surfaces, such as the soil surrounding a building, to provide support for the building, erosion control, prevention and / or repair of sinkholes, or to form a foundation structure that provides firm support and / or stabilization for buildings and other structures, and combinations thereof.

[0036] Another aspect of the invention relates to compositions, methods and systems that include one or more slurries of water, nitrogen source (e.g., urea), calcium source (e.g., CaCl2) and urease or urease-producing cells, but do not include additional or any aggregate material, such as sand, soil, dust, silt or other particles as aggregate material. Preferably, the slurry includes at least water, nitrogen source (e.g., urea), calcium source (e.g., CaCl2) and urease or urease-producing microorganisms, and may include microbial nutrients, if appropriate. This liquid slurry is sprayed, spread or otherwise placed directly on or into the aggregate material, or cast in a mold of almost any shape or configuration that includes the aggregate material. The combination of aggregate and said slurry forms a solidified object, cover or layer (or layers), such as a building foundation, a molded object, a layer covering an object or another desired form, etc. One advantage of this technology is that the aggregate material does not need to be transported, which entails savings at the same time. Preferably, the aggregate material is readily available on-site or available locally within an acceptable distance. The addition of the slurry to already existing aggregate effectively produces a solid or harder form of structure in situations where it is difficult, inefficient or impractical to transport or otherwise move the aggregate material, such as, but not limited to, situations including the construction, repair or further support of building foundations and other repairs.

[0037] Another aspect of the invention relates to compositions and methods that include the slurry of the invention in combination with an aggregate material and further include a plurality of solid structures that are hollow or otherwise lighter than the aggregate material. The resulting structure that includes the additional objects produces a solid object that is lighter than an object composed of only the aggregate material and the slurry. Alternatively, it may be desirable to increase the weight of the object by adding objects that are heavier than the aggregate material. Such heavier objects include, but are not limited to, rebar or remesh, metal moldings, reinforcements, and other heavier materials. These additional objects include, but are not limited to, plastic, wood, steel, metal, polymer, rod, ball geometric structures that may be solid, perforated, or hollow. Alternatively, additional objects may be included that have aesthetic properties, such as predefined colors, materials, functions, properties, and designs. This is advantageous when the structure retains sufficient strength for its intended purpose, such as a particular desired compressive strength, tensile strength, yield strength, ultimate strength, Young's modulus, elastic modulus, resilient strength, stiffness, hardness, toughness, stress resistance, etc., and combinations thereof, and a lightweight object is desired.

[0038] Another aspect of the present invention relates to compositions, methods and systems including various substrates combined with the slurries of the present invention. The addition of sand, fines, silt or dust (which are lighter and have smaller particles than soil or other aggregates) to urea, urease, calcium and water produces a lighter structure with equivalent or nearly equivalent bearing strength. The advantages of a lighter structure include lower production costs and higher production efficiency, as well as other benefits, such as efficiency in the manufacture and formation of the structure. Preferably, urease enzyme is used to improve the solidification of the structure compared to the use of enzyme-producing cells. In addition, the enzyme will pass through the smaller pore size of aggregate materials, which have smaller molecular structures and smaller pore sizes than cells. Also, in addition, one or more chemicals or compounds may be included to increase the density and / or weight of the liquid composition so that the composition quickly settles or adheres well to a surface (e.g., gel, foam or semi-solid).

[0039] Another preferred embodiment of the present invention includes compositions, systems and methods for forming solid or porous structures according to the present invention that are lighter in weight compared to conventional structures composed of clay or cement. Preferably, the present invention includes forming spatial voids in the solid structure as the structure hardens during manufacture. The voids can be in the form of holes, tubes, bubbles, or any other three-dimensional shape. A preformed shape composed of the same aggregate material or materials or of a different, preferably lighter material, can be immersed in the wet unhardened slurry of the present invention, either with or without aggregate material. When the slurry is fully formed around its desired shape, the resulting object will be lighter in weight than a conventionally prepared object, such as a clay brick, cement block, paving stone, composite stone, or another solid structure composed of one or more aggregate materials. The resulting solid object has increased strength, new or improved aesthetic or performance features, additives, or a combination thereof.

[0040] Another preferred feature of the present invention includes compositions, systems and methods for forming a protective layer or cover for a solid structure. Preferably, the slurries of the present invention fill and seal pores in a solid structure (e.g., fabrics impregnated with one or more of microorganisms, nutrients, substrate materials, nucleation sites) to provide an effective barrier against liquids (e.g., water), gases (e.g., contaminants) or other substances that may permeate or contaminate the solid structure. Such compositions can be used for erosion control and structural support.

[0041] Another aspect of the invention includes compositions, systems and methods for dust control, for example, of sidewalks, piles, cliffs, roadways and other large surfaces. The slurries of the invention can replace oil and other dust control compositions currently used on dirt, gravel and other road surfaces to minimize the amount of dust generated by vehicles. The slurries of the invention can be sprayed or evaporated from trucks or other vehicles as liquids, or can be applied (e.g., sprayed) onto surfaces as dry compositions that are activated when wet and form a hardened crust on the road or other surface. The slurries, which contain nutrients in addition to substrate and live urease-producing microorganisms, cover the road surface with a self-regenerating crust. The initial application may contain the microorganisms, which may optionally be included in subsequent applications that may contain only substrate material. When a vehicle passes over the road, the crust may be damaged by the weight of the vehicle, but the crust is reformed and repaired by the presence of the live slurry. Preferably, the slurries of the invention for dust control contain no aggregates or only aggregates with a diameter of 0.5 mm or less.

[0042] Another embodiment of the present invention involves adding the slurry of the present invention, with or without aggregate, to conventional processes for the production of building materials such as clay bricks, cement blocks, paving stones and other materials. The slurry additive may be included as desired at 0.0001 percent to 99 percent of the dry weight of the resulting product, or may be empirically determined from the type of aggregate used. Preferably, the slurry additive is 1 to 50 percent, 2 to 75 percent, 30 to 60 percent, 25 to 80 percent, 10 to 25 percent, or any combination thereof, by dry weight.

[0043] Another aspect of the invention involves the formation of a slurry of the invention that will solidify in a predetermined time. Preferably, the slurry contains a predetermined amount of a nitrogen source and a calcium source as substrates and a predetermined amount of enzyme that will solidify within a desired time frame. The solidification conditions may include the temperature used, which may be included in the calculation to determine the solidification time, preferably experimentally or empirically.

[0044] The following examples illustrate aspects of the present invention and should not be construed as limiting the scope of the invention. EXAMPLES

[0045] Example 1 Surface mine dust abatement is required by MSHA (the mining equivalent of OSHA) regulations. Current methods used in the industry include the use of various polymers or chemicals, with continuous spray applications of water, oil and other dust suppression liquids being the most common. The objective of surface mine dust suppression is to make fine dust (a by-product of aggregate mining) heavier than air to prevent respiratory and visual hazards. In accordance with the present invention, microorganisms are applied with nutrients and / or transport materials or concomitantly with any conventional treatment for such dust suppression and / or surface cementation, including the production of calcite cement (CaCO3) in combination with urea (nitrogen / carbon) and a calcium source. Cells and / or nutrient materials are included in the initial application and optionally in subsequent or subsequent applications. Preferably, the application is a lightweight material that readily undergoes cementation using the same urease producing bacterial strains used to form bricks, paving stones and other solid moldings. Alternatively, cyanobacteria, which are photosynthetic microorganisms that fix nitrogen from the atmosphere, can be used to replace or in addition to the urease-producing bacteria, thereby reducing the need for nutrient inputs.

[0046] Example 2 The recovery system seeks to address (a) returning the wastewater to a living state for reuse as influent (water becomes a capital expense rather than a consumable material) and (b) extraction of commercially valuable by-products from the wastewater stream. Preferably, the biocementation process of the present invention is useful for the primary production of the by-product as an output, for example using urease producing microorganisms for the manufacture of ammonia / ammonium and / or free calcite. A by-product is a surplus material that can be reduced through optimization and / or taken into account in influent formulation. Ammonia as a recoverable by-product has commercial value in both fertilizer and alternative fuel applications.

[0047] There are at least two methods for ammonia extraction. First, clinoptilolite mineral aggregate, a granular zeolite, is used as an air filter for the extraction of ammonia gas and as a liquid filter for the extraction of ammonium from wastewater. Zeolites saturated with ammonia have potential applications as fertilizers, fertilizer additives and / or fertilizer ingredients. Second, an electrode-based system is used for ammonia / ammonium water conversion as a hydrogen fuel source for electricity production.

[0048] The effluent, either further treated or untreated, is a fuel source for other ammonia-based energy production technologies, as well as reuse technologies, including reuse of water, calcite and by-products.

[0049] Settling tanks, mesh filters, fabric and / or liquid centrifuges are used to remove the free calcite in solution, preferably leaving the microorganisms behind. This material is the inoculum for new biocement formation and fertilizer applications (calcium is utilized for plant cell wall formation and the microorganisms for soil denitrification).

[0050] Example 3 Biologically formed microcrystalline calcium carbonate was produced using a urease-producing microorganism (S. pasteurii) grown in a liquid fermentation medium containing urea. The medium was stirred to produce a uniform suspension. At the later growth stage of the culture, calcium ions were added in the form of calcium chloride solution to a molar equivalent of urea to saturation. The urease activity hydrolyzes urea (2NH2CO) to ammonium (NH4) and carbon (C). The carbon combines with calcium (Ca) to produce calcium carbonate (CaCO3). The calcium carbonate crystals formed were in the size range of 50 μm to 0.1 μm and were generally "regular" (e.g., spherical) in shape. The calcium carbonate was separated from the solution using one or more of centrifugation, settling tanks, liquid centrifuges or decanting. The method was carried out as a batch process and also as a continuous production line.

[0051] A variation of this method was used to increase particle size by forming agglomerates with fine aggregate material, which also improved liquid-solid separation. In this variation, fine aggregate of size 70 mesh scale was added to the solution during the fermentation process in an amount that did not exceed the stirring capacity to keep the fine aggregate in suspension. After the addition of calcium ions, the calcite bound to the fine aggregate, binding them together to produce larger, heavier particles.

[0052] Example 4 The method of Example 3 is carried out under co-culture of a second organism, Delaya venusta. The co-culture process is constructed by circulating the medium for one fermentation in one reactor, for alternating fermentations in one reactor, or between two separate fermentations in separate reactors, either liquid-state reactors (e.g., batch, fed-batch or continuous) or solid-state reactors, such as aggregate units (e.g., bricks).

[0053] Other aspects and uses of the present invention will be apparent to those skilled in the art from consideration of the specification and practice of the invention disclosed herein. All references cited herein, including all publications, U.S. and foreign patents and patent applications, are specifically and entirely incorporated by reference. The term comprising, wherever used, is intended to include the terms consisting of and consisting essentially of. Furthermore, the terms comprising, including and containing are not intended to be limiting. It is intended that the specification and examples be considered as exemplary only, with a true scope and spirit of the invention being indicated by the appended claims.

Claims

1. A composition comprising a support material to which urease-producing cells or urease-producing cell spores have been applied and a transport medium.

2. 10. The composition of claim 1, wherein the support material comprises organic or inorganic matter, rock, glass, wood, paper, metal, plastic, polymer, fiber, mineral, or a combination thereof.

3. 10. The composition of claim 1, which is a liquid or a dry powder.

4. 10. The composition of claim 1, wherein the support material is in the form of beads, grains, rods, strands, fibers, flakes, fibres, crushed or crushed stone, crystals or combinations thereof.

5. 10. The composition of claim 1, wherein the support material is sand, glass, fiberglass, paper, ash, wood, metal, waste material or a combination thereof.

6. 2. The composition of claim 1, wherein the urease-producing cells or urease-producing cell spores comprise yeast, algae, bacteria or eukaryotic cells or cell spores.

7. 7. The composition of claim 6, wherein the bacterium is Sporosarcina pasteurii, Sporosarcina ureae, Proteus vulgaris, Bacillus sphaericus, Myxococcus xanthus, Proteus mirabilis, Helicobacter pylori, or a variant, serotype, mutant or combination thereof.

8. 7. The composition of claim 6, wherein the yeast, algae, bacterial or eukaryotic cell or cell spore is genetically engineered.

9. 10. The composition of claim 1, wherein the support material and the cells are bound by hydrophobic bonds, hydrophilic bonds, ionic bonds, non-ionic bonds, covalent bonds, van der Waals forces, or combinations thereof.

10. 10. The composition of claim 1, wherein the support is at least partially covered by a film that promotes attachment of urease-producing cells.

11. 11. The composition of claim 10, wherein the film comprises a polymer or cell nutrients.

12. 10. The composition of claim 1, further comprising a colorant.

13. 10. The composition of claim 1, comprising a distinguishing agent or detectable marker.

14. 10. The composition of claim 1, wherein the transport medium comprises components for cell growth.

15. 15. The composition of claim 14, wherein the components for cell growth include one or more of water, nutrients, vitamins, minerals, amino acids, proteins, oils, fatty acids, sugars and polysaccharides.

16. A kit for producing a solid molded product, comprising: The composition of claim 1 . a second composition comprising nutrients for the growth of urease producing cells and / or the germination of cell spores, a plurality of form sets, each set enclosing at least one solid molding shape and including one or more porous panels; and a third composition comprising calcium, nitrogen, or both calcium and urea; Including the kit.

17. 17. The kit of claim 16, wherein the solid molding is rectangular, square, circular, oval or irregularly shaped.

18. 17. The kit of claim 16, wherein the solid molding is a block, board, brick, paving stone, panel, tile or veneer.

19. 20. The kit of claim 18, wherein the block is a concrete masonry block, a cinder block, a foundation block, a breeze block, a hollow block, a solid block, a besser block, a clinker block, a high or low density block, or a cellular block.

20. 17. The kit of claim 16, wherein the nutrients include amino acids, proteins, polysaccharides, fatty acids, vitamins and minerals.

21. A method for producing a solid molded product, comprising the steps of: mixing the composition of claim 1 with aggregate material and water to form a mixture, wherein the aggregate material is composed primarily of particles less than 50 mm in diameter; distributing the mixture into a plurality of forms, each form including at least one porous panel; adding a second composition to the mixture, the second composition comprising nutrients that promote the growth of urease producing cells; adding a third composition to the mixture, the third composition being a liquid containing calcium; incubating the mixture for a period of time to allow covalent bonds to form between the particles; and Removing the solid molding from the mold A method comprising:

22. 22. The method of claim 21, wherein the aggregate material comprises rock, glass, wood, paper, metal, plastic, polymer, mineral or combinations thereof.

23. 22. The method of claim 21, wherein the mixing step comprises spraying the composition as a liquid onto the aggregate material.

24. 22. The method of claim 21, wherein the mold is substantially immersed during the incubating step.

25. 25. The method of claim 24, wherein air is bubbled into the immersed form.

26. 22. The method of claim 21, wherein additional third composition is repeatedly added to the mixture during the incubating step, drained through a bottom panel, and optionally recycled.

27. 22. The method of claim 21, wherein the incubating step is carried out under ambient conditions.

28. 22. The method of claim 21, wherein the incubating step is carried out at between 5°C and 50°C.

29. 22. The method of claim 21, wherein the third composition comprises calcium chloride, calcium acetate, calcium phosphate, calcium carbonate, calcium lactate, calcium nitrate or a calcium salt.

30. 22. The method of claim 21, wherein the pH of the mixture is monitored during the incubating step.

31. 22. The method of claim 21, wherein the solid molding is a block, board, brick, paving stone, panel, tile or veneer.

32. 22. The method of claim 21, wherein the mixture further comprises fibers or nanofibers.

33. 33. The method of claim 32, wherein the fibers or nanofibers are comprised of wood, glass, plastic, metal or polymer.

34. 22. The method of claim 21, wherein additional nutrients are added during the incubating step.

35. 22. The method of claim 21, wherein the solid molding removed from the mold is dried.

36. A composition comprising urease producing cells or urease producing cell spores encapsulated, microencapsulated or coated with a nutrient, protein, polysaccharide, polymer or other medium.

37. 37. The composition of claim 36, wherein the nutrient medium further comprises additional urease-producing cells or urease-producing cell spores.

38. A composition comprising urease and / or urease-producing cells, a nitrogen source, a calcium source and water.

39. 40. The composition of claim 38, further comprising an aggregate material having an average diameter of less than 0.05 mm.

40. 40. The composition of claim 38, further comprising an aggregate material having an average diameter between 1 nm and 40 nm.

41. 40. The composition of claim 38, further comprising an aggregate material having an average diameter of 1.0 mm to 50 mm.

42. 39. The composition of claim 38, wherein the nitrogen source is urea and the calcium source is calcium chloride.

43. 40. The composition of claim 38, comprising an amount of urease and / or urease producing cells, an amount of nitrogen source, an amount of calcium source and an amount of water that provides for solidification of the composition within a set time.

44. 40. The composition of claim 38, comprising urease producing cells and further comprising nutrients that promote cell growth.

45. 40. A method of layering a solid object with calcium carbonate, comprising contacting the solid object with the composition of claim 38 and promoting the formation of calcium carbonate.

46. 46. ​​The method of claim 45, wherein the contacting step comprises spraying the composition onto a solid object and / or maintaining a desired vapor pressure.

47. 46. ​​The method of claim 45, wherein the contacting step comprises immersing the solid object in the composition, spraying the solid object with the composition, misting the solid object with the composition, or exposing the solid object to steam along with the composition.

48. 40. A method of dust control comprising spraying the composition of claim 38 onto a surface.

49. 49. The method of claim 48, wherein the sprayed surface is more resistant to erosion compared to an unsprayed surface.

50. 49. The method of claim 48, wherein the surface is a sidewalk, a pile, a cliff, or a roadway.

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