Improved concrete composition and method for producing the same

Abstract

The present invention relates to a composition for improving concrete using microorganisms producing biochemical substances and / or byproducts synthesized by microorganisms. The present invention also relates to a method for enhancing the performance of concrete using the microbial strains and / or their byproducts.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to U.S. Provisional Patent Application No. 62,979,161, filed on February 20, 2020, the entire contents of which are incorporated herein by reference. Background Art

[0003] Concrete is one of the most common building materials, far more common than steel, plastic or wood. Concrete is currently produced by mixing cement, sand and water with added chemicals that change the concrete's properties. For example, the additives can be chemical surfactants that act as plasticizers. Plasticizers are particularly useful in producing high-strength concrete and fiber-reinforced concrete.

[0004] Increasing the water content in a concrete mix reduces concrete strength. For every 1% increase in water content, the ultimate strength of concrete decreases by 5%. However, if there is not enough water in the concrete mix, the concrete will not flow or be processed into the desired form.

[0005] Concrete plasticizers are added to concrete to reduce the amount of water required for processing. Plasticizers are usually added at concentrations of 0.1% to 0.4%. With these concentrations, the amount of water required to maintain concrete plasticity is reduced by 5% to 15%.

[0006] The cultivation of microorganisms (such as bacteria, yeast or fungi) is of great significance for the production of a variety of useful biological agents. Microorganisms play a vital role in, for example, the food industry, pharmaceutical industry, agriculture, mining, environmental remediation and waste management. In many industrial fields, the application of microorganisms has great potential.

[0007] The interest in microbial biosurfactants has steadily increased due to their diversity, environmental friendliness, selectivity, performance under extreme conditions, and potential applications in environmental protection. Biosurfactants are chemical substances of microbial origin that can reduce the interfacial tension between phases. Sophorolipids (SLPs) are biosurfactants belonging to the class of glycolipids produced by non-pathogenic yeasts. SLPs can be used in agriculture, food preservation, biomedicine, cosmetics, and other industries. Summary of the invention

[0008] The present invention provides unique and advantageous uses of microorganisms and their growth byproducts (such as biosurfactants). In a specific embodiment, the present invention provides a microbial-based product and its use in an improved concrete composition.

[0009] Specifically, in preferred embodiments, the present invention provides cost-effective, environmentally friendly methods for improving various properties of concrete. Advantageously, these methods can be implemented in a wide range of environmental conditions, including underwater and in areas with significant temperature changes.

[0010] In certain embodiments, the present invention provides compositions for improving various properties of concrete by using microorganisms and / or their growth byproducts. Microorganisms and / or their growth byproducts can be added to water, cement and aggregates and other desired ingredients for concrete products during the initial mixing of concrete. Alternatively, microorganisms and / or growth byproducts can be used at any time after the concrete begins to be poured, such as after the concrete is fully set.

[0011] In certain embodiments, the present invention utilizes yeast strains and / or their growth byproducts. In one embodiment, the microorganism used in the compositions and methods of the present invention is a yeast that produces a biosurfactant. For example, the present invention provides a microorganism-based product comprising cultured Bombus spp. Stamosaccharomyces ATCC22214 and / or the growth products of the microorganism. In addition, the present invention provides a microorganism-based product comprising cultured abnormal Wickham yeast and / or its growth byproducts. The microorganism can be in different growth phases, including a vegetative phase or a spore form.

[0012] In certain embodiments, the present invention further comprises one or more additional ingredients, for example, accelerators, binders, corrosion inhibitors, air entraining agents, crystallization admixtures, pigments, plasticizers, superplasticizers, pumping aids, retarders, water reducers, shrinkage reducers, hydration control admixtures, alkali-silica reaction inhibitors, moisture-proof admixtures, permeation-reducing admixtures, gas-generating admixtures, anti-dispersion admixtures, foaming admixtures and / or workability-enhancing admixtures.

[0013] In certain embodiments, the present invention provides methods for improving the properties of concrete, wherein a composition comprising a microbial culture and / or microbial growth byproducts according to the present invention is added to a concrete mix and / or applied to the surface of curing or cured concrete. The properties of concrete can be improved by, for example, reducing the amount of water in the concrete composition, increasing the plasticity of the concrete, reducing the porosity of the concrete, changing the curing time of the concrete, and / or increasing the tolerance to biotic and / or abiotic stresses. DETAILED DESCRIPTION

[0014] The present invention provides advantageous uses of microorganisms and their growth byproducts (such as biosurfactants). In certain embodiments, the present invention provides a microbial-based product and its use in improved concrete compositions.

[0015] In specific embodiments, the methods and compositions described herein utilize microorganisms and / or microbial growth byproducts as admixtures to concrete. In one embodiment, the microorganism used in the methods of the present invention is a yeast that produces a biosurfactant.

[0016] In certain embodiments, the present invention utilizes yeast strains and / or growth byproducts thereof. The present invention provides, for example, microbial-based products comprising, for example, cultured abnormal Wickham yeast and / or growth byproducts thereof. In addition, the present invention provides microbial-based products comprising cultured Starmerella branch yeast, preferably Bombus spp. Starmerella (Candida), and / or growth byproducts thereof.

[0017] In a preferred embodiment, the present invention provides a method for improving one or more properties of concrete by applying one or more microorganisms and / or microbially derived biosurfactants to the concrete. In a specific embodiment, the method comprises applying one or more biosurfactants derived from a yeast strain such as Bombyx mossima or Wickhamia anomala to the concrete.

[0018] In certain embodiments, the method may further include using the microorganisms themselves and / or their byproducts together with one or more admixtures conventionally used in concrete.

[0019] In specific embodiments, the methods and compositions described herein utilize microbial growth byproducts as use products during or after concrete curing. The microorganisms and / or byproducts may penetrate into the concrete or remain on the surface of the concrete.

[0020] In one embodiment, the composition according to the present invention is obtained by a culture process ranging from small scale to large scale. These culture methods include, but are not limited to, submerged culture / fermentation, surface culture, solid state fermentation (SSF) and combinations thereof.

[0021] Advantageously, the present invention may not release large amounts of inorganic compounds into the environment during use. In addition, the compositions and methods utilize biodegradable and safe, non-toxic ingredients. Therefore, the present invention may be used as a "green" treatment for all possible coatings (and other applications).

[0022] Selected Definitions

[0023] As used herein, a "microbial-based composition" refers to a composition that includes components produced by the growth of a microorganism or other cell culture. Thus, a microbial-based composition may include the microorganism itself and / or microbial growth byproducts. The cells may be in the vegetative stage or in spore form, or a mixture of the two. The cells may be planktonic or in biofilm form, or a mixture of the two. Growth byproducts may be, for example, metabolites, cell membrane components, expressed proteins, and / or other cell components. The cells may be intact or lysed cells. In some embodiments, the cells are present in a microbial-based composition together with the liquid culture medium in which they are grown. The cells may be present in a composition at, for example, at least 1×10 4 , 1×10 5 , 1×10 6 , 1×10 7 , 1×10 8 , 1×10 9 , 1×10 10 or 1×10 11 or more cells are present.

[0024] The present invention further provides "microorganism-based products", which can be used in practice to obtain expected results. Microorganism-based products can be microorganism-based compositions harvested from microorganism culture processes. Alternatively, microorganism-based products can contain other ingredients that have been added. These additional ingredients can include, for example, buffers, suitable carriers (such as water), added nutrients for supporting further growth of microorganisms, and / or reagents that promote tracking of microorganisms and / or compositions in the environment in which they are used. Microorganism-based products can also include mixtures of microorganism-based compositions. Microorganism-based products can also include one or more components of microorganism-based compositions that have been processed in some way, such as but not limited to filtration, centrifugation, lysis, drying, purification, etc.

[0025] As used herein, an "isolated" or "purified" biosurfactant or other microbial-derived biochemical is substantially free of other compounds, such as cellular material, with which it is associated in nature. As used herein, "isolated" means that the strain is removed from the environment in which it naturally occurs. Thus, an isolated strain may exist, for example, as a biologically pure culture or spore (or other form of strain).

[0026] "Metabolite" refers to any substance produced by metabolism (e.g., a growth byproduct) or a substance necessary to participate in a specific metabolic process. Examples of metabolites include, but are not limited to, biosurfactants, biopolymers, enzymes, acids, solvents, alcohols, proteins, vitamins, minerals, trace elements, and amino acids.

[0027] "Modulate" means to change (increase or decrease). Such changes are detected by standard art-known methods, such as those described herein.

[0028] The scope provided herein is understood to represent all values ​​in the range. For example, the scope of 1-50 is understood to include all decimal values ​​centered between any number, combination of numbers or subranges and the above-mentioned integers selected from 1,2,3,4,5,6,7,8,9,10,11,12,13,14,15,16,17,18,19,20,21,22,23,24,25,26,27,28,29,30,31,32,33,34,35,36,37,38,39,40,41,42,43,44,45,46,47,48,49 or 50, such as 1.1,1.2,1.3,1.4,1.5,1.6,1.7,1.8 and 1.9. Special consideration is given to " nested subranges " starting from any end point of scope in the subrange. For example, nested sub-ranges of the exemplary range of 1-50 may include 1-10, 1-20, 1-30, and 1-40 in one direction, or 50-40, 50-30, 50-20, and 50-10 in the other direction.

[0029] "Decrease" refers to a negative change of at least 1%, 5%, 10%, 25%, 50%, 75% or 100%.

[0030] An "increase" is a positive change of at least 1%, 5%, 10%, 25%, 50%, 75% or 100%.

[0031] "Reference" refers to a standard or control condition.

[0032] As used herein, "biofilm" is a complex aggregate of microorganisms (e.g., bacteria) in which cells adhere to each other and / or to a surface using an extracellular polysaccharide matrix. Cells in a biofilm are physiologically distinct from planktonic cells of the same organism, which are single cells that can float or swim in a liquid medium.

[0033] As used herein, "surfactant" refers to a compound that reduces the surface tension (or interfacial tension) between two liquids or between a liquid and a solid. Surfactants are used as detergents, wetting agents, emulsifiers, foaming agents and / or dispersants, etc. Surfactants produced by microorganisms are called "biosurfactants."

[0034] As used herein, "biotic stress" is stress on a composition and / or object caused by living organisms, such as bacteria, fungi, or barnacles. "Biological stressors" are different from "abiotic stressors" because "abiotic stressors" are the result of non-living factors, such as ultraviolet radiation, freeze-thaw cycles, or salt water.

[0035] As used herein, "accelerators", "cement quick-setting agents", "accelerators" and "concrete quick-setting agents" are ingredients used in concrete, cement, mortar, plaster or screed that accelerate the setting time of the concrete composition. Accelerators can be used to eliminate the effects of cold weather (whether it is raining or not) that delay the setting time or damage the concrete. Accelerators include, but are not limited to, calcium nitrate, calcium nitrite, calcium formate, calcium chloride, triethanolamine and sodium thiocyanate.

[0036] As used herein, "binders," "adhesives," "resins," and "epoxies" are ingredients used in concrete that bond concrete to existing structures, such as concrete, stucco, and gypsum. Without a binder, new concrete cannot adhere to an already cured concrete structure. Binders include, but are not limited to, polyvinyl chloride, polyvinyl acetate, acrylics, and butadiene-styrene copolymers.

[0037] As used herein, "corrosion inhibitors" are ingredients used in concrete that can prevent premature failure of reinforced concrete by limiting the reaction of chlorides and atmospheric CO2 with cement. Chlorides from sources such as deicing salts penetrate buried steel reinforcement, causing rust. Corrosion inhibitors include, but are not limited to, amino alcohols, calcium nitrite, and sodium monofluorophosphate.

[0038] As used herein, "air entraining agents" are compounds used in concrete to produce microscopic bubbles in the composition. Air entraining agents promote the formation of air pockets in concrete by allowing air to expand and condense during temperature cycles, thereby increasing durability, improving the workability and cohesion of concrete, and reducing segregation and bleeding of concrete. Air entraining agents include salts of wood resins, synthetic detergents, salts of sulfonated lignin, salts of petroleum acids, salts of proteinaceous substances, fatty acids and resin acids and their salts, alkylbenzene sulfonates, and salts of sulfonated hydrocarbons.

[0039] As used herein, a "crystalline admixture" or "permeability reducer" reduces the permeability of concrete to which it is added. In some cases, with the addition of an appropriate amount of a crystalline admixture, concrete may become virtually impermeable. Crystalline admixtures include, but are not limited to, latex and calcium stearate.

[0040] As used herein, "plasticizers," "superplasticizers," "workability enhancing admixtures," and "water reducers" are added to concrete to reduce the amount of water required for working, increase fluidity, and / or reduce the water-cement ratio. Chemical plasticizers include, but are not limited to, calcium lignin sulfonates, polyglucose esters, carbohydrates, and hydroxylated carboxylic acids. There are also superplasticizers, including modified lignin sulfates, sulfonated naphthaldehydes, which are superior to plasticizers in their ability to allow concrete workability, with up to 30% less water.

[0041] As used herein, "pumping aids" act as lubricants that allow concrete to flow easily through pipes, channels, barrels, or other objects used for concrete transportation or holding rooms. Pumping aids include, but are not limited to, organic and synthetic polymers, organic flocculants, paraffin organic emulsions, coal tar pitch, acrylic resins, bentonite and pyrogenic silica, and slaked lime.

[0042] As used herein, a "retarder" is a substance that reduces the time it takes for concrete to set. Retarders include, but are not limited to, lignin, borax, sugar, tartaric acid, and salt. Similarly, a "hydration control admixture" can also reduce the time it takes for concrete to set, but hydration control admixtures are generally more forgiving after the evaporation process begins. A hydration control admixture can be likened to a time-release capsule.

[0043] According to the present invention, the "shrinkage reducing agent" limits the shrinkage of the concrete when the concrete is set. The shrinkage reducing agent includes but is not limited to polyoxyethylene alkyl ether and propylene glycol.

[0044] As used herein, "alkali-silica reaction inhibitors" inhibit expansion that may occur over the life of the concrete by inhibiting the reaction between alkaline cement paste and silica. Sodium silicate is produced in the reaction, which in turn produces a hygroscopic gel that increases in volume when exposed to water. Alkali-silica reaction inhibitors include, but are not limited to, barium salts, lithium nitrate, lithium carbonate, and lithium hydroxide.

[0045] As used herein, "moisture-proofing" agents prevent or stop moisture from penetrating into concrete. Moisture-proofing is generally used to prevent moisture from the surrounding environment from penetrating into concrete. Such sources of moisture may be soil or sand surrounding basement walls, concrete slabs, or sidewalk concrete. Moisture-proofing is generally applied as an exterior coating to concrete, rather than as an additive mixed into the overall concrete composition. Some materials that can be used to moisture-proof concrete include calcium oleate or ammonium stearate soaps, calcium oleate or ammonium oleate soaps, butyl stearate, and various petroleum products.

[0046] As used herein, a "waterproofing" agent is similar to a moisture barrier admixture, but is intended to prevent liquid water penetration, especially in environments with high groundwater levels, frequent flooding, or in pools. Waterproofing materials are also used on the exterior surfaces of concrete. Some materials that can be used to waterproof concrete include rubberized asphalt coatings, bentonite, rubber coatings, and polyurethane coatings.

[0047] As used herein, "gassing" agents are substances used to expand concrete before it is cured. The most common gassing admixtures are aluminum powder, activated carbon, and hydrogen peroxide. These added chemicals form hydrogen bubbles in the concrete. In addition to preventing settling on the concrete, the presence of hydrogen prevents bleeding in the concrete. Bleeding in concrete is when water is pushed upward and the denser cement particles sink. Gassing admixtures can also create lightweight concrete.

[0048] As used herein, "anti-dispersion" agents are typically added to concrete intended for underwater use to mitigate the effects of water pressure and friction caused by the movement of water. Anti-dispersion admixtures can improve the cohesion of concrete and prevent the loss of cement paste. The concrete can be self-compacting concrete. Cellulose and acrylic polymers can be used in anti-dispersion compositions.

[0049] As used herein, a "foaming" agent is a substance that is produced by adding surfactants or other chemicals, such as hydrolyzed proteins, to the concrete composition to produce a lighter, less dense concrete. Concrete containing a foaming mixture is typically used for void filling, as opposed to structural support.

[0050] As used herein, "aggregate" or "construction aggregate" constitutes the three basic ingredients for making concrete with water and cement. Aggregate is a granular material. Aggregate usually constitutes the main part of the concrete composition, including sand, stone, gravel, glass, silica, blast furnace slag, recycled concrete, rock or other related ingredients.

[0051] The transitional term "comprising," which is synonymous with "including" or "containing," is inclusive or open-ended and does not exclude additional, unrecited elements or method steps. In contrast, the transitional phrase "comprising" excludes other elements, steps, or ingredients not specified in the claim. The transitional phrase "consisting essentially of" limits the scope of a claim to the specified materials or steps "and those that do not materially affect the basic and novel characteristics of the claimed invention." Use of the term "comprising" contemplates embodiments "consisting of" or "consisting essentially of" the recited elements.

[0052] Unless specifically stated or obvious from the context, the term "or" as used herein should be understood to be inclusive. Unless specifically stated or obvious from the context, the terms "a", "an" and "the" as used herein should be understood to be singular or plural.

[0053] Unless otherwise specified or apparent from the context, the term "about" as used herein is understood to be within the normal tolerance range in the art, such as within 2 standard deviations of the mean. About can be understood to be within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05% or 0.01% of the stated value.

[0054] The description of a list of chemical groups in any definition of a variable herein includes definitions of that variable as any single group or combination of listed groups. The description of an embodiment of a variable or aspect herein includes that embodiment as any single embodiment or in combination with any other embodiment or portion thereof.

[0055] Yeast strain according to the invention

[0056] In a preferred embodiment, the present invention utilizes yeast that produces biochemical substances. Yeast can be a natural or genetically modified microorganism. For example, yeast can be transformed with a specific gene to display a specific characteristic. Yeast can also be a mutant of a desired strain. As used herein, "mutant" refers to a strain, genetic variant or subtype of a standard bacterium, wherein the mutant has one or more genetic variations (e.g., point mutations, missense mutations, nonsense mutations, deletions, duplications, frameshift mutations or repeated amplifications) compared to the standard bacterium. Methods for preparing mutants are well known in the microbiology field. For example, ultraviolet mutagenesis and nitrosoguanidine are widely used for this purpose.

[0057] Examples of yeast and fungal species suitable for use in the present invention include, but are not limited to, Aspergillus, Aspergillus, Mortierella (e.g., Black Saccharomyces), Blakeslea, Candida (e.g., Candida albicans, Candida albicans), Cryptococcus, Debaryomyces (e.g., Debaryomyces hansenii), Entomophthora, Fusarium, Hansenula (e.g., Hansenula viticola), Hansenula, Issatchenko, Kluyveromyces, Mortierella, Mucor (e.g., Mucor piriformis), Meyer yeast (e.g., Meyer yeast guillemot), Penicillium, Pythium, Phycomyces, Pichia (e.g., Pichia anomala, Pichia guillemot, Pichia occidentalis, Pichia kudrida), yeast (e.g., Pseudomonas aphidomyces), Rhizopus, yeast (Saccharomyces cerevisiae, Saccharomyces boulardii sequela), Torulella cerevisiae), Starmosaccharomyces (e.g., Starmosaccharomyces bumblebee), Torulopsis, Chytridiomyces, Trichoderma (e.g., Trichoderma reesei, Trichoderma harzianum, Trichoderma viride), Ustilago (e.g., Ustilago maydis), Wickhamia (e.g., Wickhamia anomala), Pseudomonas, and Zygosaccharomyces (e.g., Zygosaccharomyces bailii).

[0058] In some embodiments, yeast is "killer yeast", i.e., a yeast strain characterized by secreting toxic proteins or glycoproteins, which has immunity itself. Exotoxins secreted by killer yeast can kill other strains such as yeast, fungi or bacteria. Killer yeast can include but is not limited to multiple species in the genus Wickham yeast (abnormal Wickham yeast), Pichia, Hansenula, yeast, Hansenula (e.g., Hansenula spores in grape juice), corn smut, Hansen Debaryomyces, multiple species in the genus Candida (white Candida, wrinkled Candida, tropical Candida, Candida lipolytica, white Torulopsis), multiple species in Starmoyces (bombus-born Starmoyces), Cryptococcus, Kluyveromyces, Torulopsis, Ustilago, Willems, Zygosaccharomyces and others.

[0059] In certain embodiments, the microorganism is a variety of yeasts in Stamosaccharomyces and / or a variety of yeasts in the genus Candida, Candida apicola, Candida batistae, Candida floricola, Candida riodocensis, Candida stellate and / or Candida kuoi. In a specific embodiment, the microorganism is Stamosaccharomyces bumblebee, such as strain ATCC 22214.

[0060] In certain preferred embodiments, the microorganism is Wickhamella anomala.

[0061] According to the present invention, other microbial strains may be used, including other fungal strains such as those that are capable of accumulating large amounts of glycolipid biosurfactants or other useful metabolites such as carbohydrates, polyols, lipids, esters and / or proteins. Other metabolites and / or microbial components useful according to the present invention include mannoproteins, β-glucans and other substances with bioemulsification and surface / interfacial tension reducing properties.

[0062] Growth of the yeast according to the invention

[0063] The present invention utilizes methods of culturing microorganisms and producing microbial metabolites and / or other microbial growth byproducts. Microbial culture systems typically use submerged culture fermentation, but surface culture and mixing systems may also be used.

[0064] As used herein, "fermentation" refers to the growth of cells under controlled conditions. Growth can be carried out in an aerobic environment or an anaerobic environment. Unless the context requires otherwise, the term is intended to include both the growth phase and the product biosynthesis phase of the process.

[0065] As used herein, "liquid medium", "culture liquid medium" or "fermentation broth" refers to a medium containing at least nutrients. If a liquid medium is mentioned after a fermentation process, the liquid medium may also contain microbial growth byproducts and / or microbial cells.

[0066] In one embodiment, the invention provides materials and methods for producing biomass (e.g., living cell material), extracellular metabolites (e.g., small molecules and secreted proteins), residual nutrients, and / or intracellular components (e.g., enzymes and other proteins).

[0067] The microbial growth container used according to the present invention can be any fermentation tank or culture reactor for industrial use. The terms "reactor", "bioreactor" or "fermentation reactor" used herein include a fermentation unit composed of one or more containers and / or towers or pipeline arrangements. Reactors include but are not limited to continuous stirred tank reactors (CSTRs), immobilized cell reactors (ICRs), trickle bed reactors (TBRs), bubble tower reactors, airlift fermenters, static mixers or other containers or other devices suitable for gas-liquid contact. In certain embodiments, a bioreactor may include a first growth reactor and a second fermentation reactor. Therefore, when referring to adding a substrate to a bioreactor or a fermentation reactor, it should be understood to include adding a substrate to any one or two suitable reactors.

[0068] In one embodiment, the container may have, or may be connected to, functional controls / sensors to measure important factors in the culture process, such as pH, oxygen, pressure, temperature, agitator shaft power, humidity, viscosity and / or microbial density and / or metabolite concentration.

[0069] In another embodiment, the container can also monitor the growth of microorganisms in the container (e.g., measuring the number of cells and the growth period). Alternatively, samples can be removed from the container daily and counted by techniques such as the dilution plate technique known in the art. The dilution plate method is a simple method for estimating the number of cells in a sample. The technique can also provide an index by which different environments or treatments can be compared.

[0070] In one embodiment, the method includes supplementing the culture with a nitrogen source. The nitrogen source may include, for example, potassium nitrate, ammonium nitrate, ammonium sulfate, ammonium phosphate, ammonia, urea and / or ammonium chloride. These nitrogen sources may be used alone or in combination.

[0071] The culture method can be to perform oxygenation treatment for the growing culture. In one embodiment, the slow movement of air is used to remove the low oxygen content air and introduce oxygenated air. The oxygenated air can be air replenished daily by a mechanical device, which includes an impeller for mechanical agitation of the liquid and an air sparger for providing bubbles to the liquid to dissolve oxygen into the liquid.

[0072] The method may further include supplementing the culture with a carbon source. The carbon source is typically a carbohydrate, such as glucose, sucrose, lactose, fructose, trehalose, mannose, mannitol and / or maltose; an organic acid, such as acetic acid, fumaric acid, citric acid, propionic acid, malic acid, malonic acid and / or pyruvic acid; an alcohol, such as ethanol, propanol, butanol, pentanol, hexanol, isobutanol and / or glycerol; a fat and oil, such as soybean oil, rice bran oil, olive oil, corn oil, sesame oil and / or linseed oil. These carbon sources may be used alone or in combination.

[0073] In one embodiment, the culture medium includes growth factors and micronutrients of microorganisms. In one embodiment, inorganic salts may also be included. In certain embodiments, the culture method may further include: adding additional acid and / or antimicrobial agents in the liquid culture medium before and / or during the culture process. Antimicrobial agents or antibiotics are used to protect the culture from contamination. In addition, defoamers may also be added to prevent the formation and / or accumulation of foam during gas generation during the culture process.

[0074] The pH of the mixture should be appropriate for the microorganism being studied. Buffers and pH adjusters, such as carbonates and phosphates, can be used to stabilize the pH around the preferred value. When metal ion concentrations are high, it may be necessary to use chelating agents in the liquid culture medium.

[0075] The microorganisms can grow in a planktonic form or in a biofilm form. In the case of a biofilm, the container may have a substrate on which the microorganisms can grow in a biofilm state. The system may also have the ability to apply stimuli (e.g., shear stress) to promote and / or improve biofilm growth characteristics.

[0076] In one embodiment, the method of culturing microorganisms is carried out at a temperature of about 5°C-100°C, preferably 15°C-60°C, and more preferably 25°C-50°C. In another embodiment, the culturing can be carried out continuously at a constant temperature. In another embodiment, the culturing can be carried out under temperature variation conditions.

[0077] In one embodiment, the equipment used in the method and the culture process is a sterile equipment. Culture equipment such as reactors / containers can be separate and yet connected to a disinfection unit such as an autoclave. Culture equipment can also have a disinfection unit for on-site disinfection before inoculation. Air can be disinfected by methods known in the art. For example, ambient air can pass through at least one filter before being introduced into the container. In other embodiments, the culture medium can be pasteurized, or can be selected not to be sterilized by heating, and low water activity and low pH can be used to control bacterial growth.

[0078] The biomass content of the fermentation broth may be 5 g / l-180 g / l or higher. In one embodiment, the solid content of the liquid culture medium is 10 g / l-150 g / l.

[0079] In one embodiment, the present invention further provides a method for producing microbial metabolites, including but not limited to ethanol, polyols, esters, lactic acid, β-glucan, proteins, peptides, metabolic intermediates, polyunsaturated fatty acids, biosurfactants and lipids. The metabolite content produced by the method can reach at least 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90%.

[0080] The microbial growth byproducts produced by the microorganism under study can be retained in the microorganism or secreted into the liquid culture medium. In another embodiment, the method for producing microbial growth byproducts can further include the steps of concentrating and purifying the microbial growth byproducts under study. In another embodiment, the liquid culture medium can contain compounds that stabilize the activity of the microbial growth byproducts.

[0081] In a preferred embodiment, the microbial growth byproduct is a biosurfactant. Specific biosurfactants according to the present invention include, for example, low molecular weight glycolipids (GL), lipopeptides (LP), flavin lipids (FL), phospholipids and high molecular weight polymers, such as lipoproteins, lipopolysaccharide-protein complexes and polysaccharide-protein-fatty acid complexes.

[0082] In one embodiment, the microbial biosurfactant is a glycolipid, such as a rhamnolipid (RLP), a sophorolipid (SLP), a trehalose lipid or a mannoerythritol lipid (MEL). In one embodiment, the microbial biosurfactant is a lipopeptide, such as iturin, fengycin or surfactin.

[0083] In one embodiment, the yeast-based composition comprises any mixture of these biosurfactants. Preferably, the mixture comprises sophorolipids, and optionally, one or more mannoerythritol lipids, surfactins, iturin and / or rhamnolipids.

[0084] In certain embodiments, microbial growth byproducts include carbohydrates, polyols, lipids, glycolipids, esters and / or proteins. These components may be waste byproducts produced by fermentation. Growth byproducts may also be chemicals remaining after yeast cells die.

[0085] In certain embodiments, the purified compound is at least 60wt% of the compound under investigation. Preferably, the preparation is at least 75wt% of the compound under investigation, more preferably 90wt%, and most preferably 99wt%. For example, the purified compound is at least 90wt%, 91wt%, 92wt%, 93wt%, 94wt%, 95wt%, 98wt%, 99wt% or 100wt% (w / w) of the desired compound. Purity is measured by any suitable standard method, for example, column chromatography, thin layer chromatography or high performance liquid chromatography (HPLC) analysis.

[0086] The methods and apparatus for culturing microorganisms and producing microbial byproducts can be performed in a batch, quasi-continuous, or continuous process.

[0087] In one embodiment, when the culture is completed (e.g., when the desired cell density or density of a specific metabolite in the liquid culture medium is reached), all of the microbial culture composition is removed. In the batch process, after the first batch is completed, a new batch of culture is started.

[0088] In another embodiment, only a portion of the fermentation product is removed at any one time. In this embodiment, the biomass with living cells is retained in the container as an inoculum for a new culture batch. The removed composition may or may not contain a liquid culture medium with cells. In this way, a quasi-continuous system is established.

[0089] Advantageously, the method does not require complex equipment or high energy consumption. The microorganisms under study can be cultivated and used on site at different scales, even while remaining mixed with the culture medium. Similarly, microbial metabolites can be produced in large quantities when needed.

[0090] Advantageously, microbial-based products can be produced in remote locations. Microbial growth facilities can be operated off-grid by utilizing power such as solar, wind, and / or hydroelectric power.

[0091] Preparation of yeast-based products

[0092] A yeast-based product of the present invention is simply a fermentation broth containing yeast and / or microbial metabolites produced by the yeast and / or any residual nutrients. The fermentation product can be used directly without extraction or purification. If necessary, extraction and purification can also be easily achieved using standard extraction and / or purification methods or techniques in the literature.

[0093] The yeast in the yeast-based products can be in active or inactive form, or a combination of both. Yeast-based products can be used without further stabilization, preservation, and storage. Advantageously, direct use of these yeast-based products ensures high viability of the microorganisms, reduces the likelihood of contamination by foreign matter and undesirable microorganisms, and maintains the activity of microbial growth byproducts.

[0094] In one embodiment, the first yeast fermentation product named "Star 3+" can be obtained by using cultured yeast (e.g., abnormal Wickham yeast) in a modified solid-state fermentation form. Abnormal Wickham yeast is often associated with food and grain production, and is also effectively used to produce various solvents, enzymes, toxins, triglycerides, and biosurfactants (e.g., phospholipids). The culture can be grown on a substrate with sufficient surface area, and the yeast can attach to and multiply on the substrate, such as rice, soybeans, chickpeas, pasta, oatmeal, or beans. The entire fermentation medium containing all grown yeast cells and any growth byproducts thereof (e.g., enzymes, solvents, and / or biosurfactants) can be obtained after culturing at 25-30° C. for 3-5 days. The culture can be mixed with the substrate, ground and / or micronized, and optionally dried. Among them are Star 3+ products. The composition can contain, for example, 1×10 10 -10 12 The cells can be diluted 10-fold, 50-fold, 100-fold, 500-fold or 1000-fold, etc. before mixing with other components.

[0095] In one embodiment, the yeast fermentation product can be obtained by submerged culture of abnormal Wickham yeast. The fermentation broth after culturing at 25°C-30°C for 7 days can contain a yeast cell suspension and, for example, at least 1g / L-100g / L, 2g / L-80g / L, 3g / L-60g / L, 4g / L-40g / L, 5g / L-20g / L or 6g / L-10g / L of a biosurfactant.

[0096] In one embodiment, the yeast fermentation product can also be obtained by culturing the biosurfactant-producing yeast Bombus stamina. This strain can be effectively used to produce glycolipid biosurfactants, such as SLP. The fermentation broth after culturing for 5 days at 25°C can contain a yeast cell suspension and, for example, at least 1 g / L-150 g / L, 2 g / L-120 g / L, 3 g / L-100 g / L, 4 g / L-80 g / L, 5 g / L-60 g / L or 6 g / L-50 g / L of glycolipid biosurfactant.

[0097] In specific embodiments, the biosurfactant of the subject composition comprises one or more glycolipid biosurfactants. In specific preferred embodiments, the glycolipid is a sophorolipid.

[0098] Sophorolipids are glycolipid biosurfactants produced by yeasts such as the Starmosaccharomyces branch, and in some embodiments, by abnormal Wickham yeast. SLP consists of a disaccharide sophorose connected to a long-chain hydroxy fatty acid. It may contain a partially acetylated 2-O-β-D-pyranose glucopyranose unit, connected to 17-L-hydroxyoctadecanoic acid or 17-L-hydroxy-Δ9-octadecenoic acid with a β-glycosidic bond. Hydroxy fatty acids typically contain 16 or 18 carbon atoms and may contain one or more unsaturated bonds. In addition, the sophorose residue may be acetylated at the 6-position and / or 6'-position. The fatty acid carboxyl group may be in free form (acidic or linear form (Formula 1)) or esterified internally at the 4"-position (lactone form (Formula 2)). Bombus spp. produces a special enzyme, called Bombus spp. lactone esterase, which catalyzes the esterification reaction of linear SLP to produce lactone-type SLP.

[0099]

[0100] In the formula, R 1 and R 1’ Respectively represent a saturated hydrocarbon chain or a single or multiple (especially single) unsaturated hydrocarbon chain having 8 to 20 (specifically 12 to 18, more preferably 14 to 18) carbon atoms, which may be straight or branched and may contain one or more hydroxyl groups, R 2 and R 2’ represents a hydrogen atom or a saturated alkyl functional group or a single or multiple (especially single) unsaturated alkyl functional groups having 1 to 9 (more preferably 1 to 4) carbon atoms, which may be linear or branched and may contain one or more hydroxyl groups, and R 3 , R 3’ , R 4 and R 4’ represent a hydrogen atom or an acetyl group, respectively.

[0101] Sophorolipids can be present in the form of pure substances or mixtures of fermentation products. Sophorolipids can be added to the concrete composition at a concentration of 0.001wt%-90wt% (wt%), preferably 0.01wt%-50wt%, more preferably 0.1wt%-20wt%. In another embodiment, pure SLP can be combined with an acceptable carrier, wherein the concentration of SLP can be 0.001%-50% (v / v), preferably 0.01%-20% (v / v), more preferably 0.02%-5% (v / v).

[0102] Yeast and / or liquid medium produced by the yeast growth may be removed from the growth vessel and transferred, for example, by piping, for immediate use.

[0103] The yeast fermentation product may include yeast cells and fermentation broth, or may include fermentation broth separated from yeast cells.In one embodiment, the biosurfactant or other growth byproducts in the liquid culture medium are further separated and purified from the liquid culture medium.

[0104] In other embodiments, the composition (yeast, liquid culture medium, or yeast and liquid culture medium) can be placed in a container of appropriate size, taking into account factors such as the intended use, the planned method of use, the size of the fermenter, and the available means of transportation from the microbial growth facility to the point of use. Based on the above considerations, the container holding the yeast-based composition can be 1 gallon to 2000 gallons or larger. In other embodiments, the container is 2 gallons, 5 gallons, 25 gallons, 250 gallons or larger.

[0105] In certain embodiments, the compositions of the invention have advantages over, for example, biosurfactants alone, including one or more of the following: a high concentration of mannoproteins as part of the outer surface of the yeast cell wall (mannoproteins are efficient bioemulsifiers); the presence of the biopolymer β-glucan (an emulsifier) ​​in the yeast cell wall; and the presence of biosurfactants, metabolites, and solvents (e.g., lactic acid, ethanol, ethyl acetate, etc.) in the culture.

[0106] Other chemical substances useful according to the present invention include mannoproteins, beta-glucans, ethanol, lactic acid and other metabolites having properties such as bioemulsification and surface / interfacial tension reduction.

[0107] In certain embodiments, microbial cultures and / or growth byproduct compositions can be added to concrete. These compositions can be added during the initial mixing of the concrete ingredients, or the microbial cultures and / or growth byproduct compositions can be added after the initial concrete use, i.e., during or after the curing process.

[0108] Concrete has three basic ingredients: water, cement, and aggregates. Aggregates, such as sand, stone, rock, gravel, recycled concrete, glass, or blast furnace slag, make up the bulk of concrete and provide the basic structure of concrete. In some embodiments, aggregates can include a single ingredient, such as sand, or a combination of different aggregates, such as sand, recycled concrete, and glass. Cement is used as a binder to bind the aggregates to each other. Water is added to make the mixture a uniform, workable composition. This concrete mix can be made into a variety of shapes and compositions depending on the application.

[0109] Various admixtures are added to the concrete mixture, including accelerators, binders, corrosion inhibitors, air entraining agents, crystallizing admixtures, pigments, plasticizers, superplasticizers, pumping aids, retarders, water reducers, shrinkage reducers, hydration control admixtures, alkali-silica reaction inhibitors, moisture-proof admixtures, seepage-reducing admixtures, gas-generating admixtures, anti-dispersion admixtures, foaming admixtures and / or workability-enhancing admixtures.

[0110] In certain embodiments, an accelerator is used in or in combination with the microbial culture and / or growth byproduct composition. Calcium chloride, calcium nitrate, calcium nitrite, calcium formate, calcium chloride, triethanolamine, and sodium thiocyanate can be used in concrete compositions, but typically cause corrosion problems, thus requiring the use of corrosion inhibitors.

[0111] In certain embodiments, corrosion inhibitors are used in or in combination with the microbial culture and / or growth byproduct composition. Corrosion inhibitors include, but are not limited to, amino alcohols such as 2-aminomethylpropanol; benzotriazoles such as dimethylethanolamine benzotriazole and methylbenzotriazole; calcium nitrite; and sodium monofluorophosphate.

[0112] Microbial cultures and / or growth byproducts may be used in conjunction with compositions that promote adhesion of the microbial cultures and / or growth byproducts to the surface to be treated. Adhesion promoting substances may be a component of the microbial cultures and / or growth byproducts, or may be used in sequence with yeast-based products. Adhesives, binders, resins, and epoxies are ingredients used in concrete to bond concrete to existing structures or to bond additional chemicals to concrete surfaces. Adhesives help new concrete adhere to already cured concrete structures. Adhesives that may be used in the present invention include, but are not limited to, polyvinyl chloride, polyvinyl acetate, acrylic resins, and butadiene-styrene copolymers.

[0113] In certain embodiments, air-entraining agents are used in or in combination with the microbial culture and / or growth byproduct composition. Air-entraining agents include salts of wood resins, synthetic detergents, salts of sulfonated lignin, salts of petroleum acids, salts of proteinaceous materials, fatty acids and resin acids and their salts, alkylbenzene sulfonates, and salts of sulfonated hydrocarbons.

[0114] In certain embodiments, crystallization admixtures are used in or in conjunction with the microbial culture and / or growth byproduct compositions, including, but not limited to, latex and calcium stearate.

[0115] In certain embodiments, the microbial culture and / or growth byproduct compositions of the present invention include pigments or dyes, which can provide the color of paint or other coatings, but can additionally protect surfaces or objects from, for example, ultraviolet radiation. The pigments or dyes can be natural, synthetic, inorganic or organic. The pigments or dyes can be selected from, for example, titanium dioxide, zinc oxide, zinc yellow, yellow dyes, benzidine yellow, chromium oxide green, phthalocyanine green, phthalocyanine blue, ultramarine blue, vermilion, pigment brown 6, red 170, dioxazine purple, carbon black, ferrous oxide, quartz sand (SiO ), talc, barite (BaSO ), kaolin and limestone (CaCO ).

[0116] In certain embodiments, plasticizers, superplasticizers, workability-enhancing admixtures, and water reducers may be added to the microbial culture and / or growth byproduct composition, or used in combination therewith. Chemical plasticizers include, but are not limited to, calcium lignin sulfonate, polyglucose esters, carbohydrates, and hydroxylated carboxylic acids. There are also superplasticizers, including, for example, modified lignin sulfates, sulfonated naphthalene formaldehydes, which are superior to plasticizers in their ability to allow for increased workability of concrete.

[0117] In certain embodiments, pumping aids may be added to or used in conjunction with the microbial culture and / or growth byproduct composition. Pumping aids include, but are not limited to, organic and synthetic polymers, organic flocculants, paraffin organic emulsions, coal tar pitch, acrylic resins, bentonite and pyrogenic silica and slaked lime.

[0118] In certain embodiments, retarders and hydration control admixtures may be added to or used in conjunction with the microbial culture and / or the growth byproduct composition. Retarders include, but are not limited to, lignin, borax, sugars, tartaric acid, and salts.

[0119] In certain embodiments, shrinkage reducing agents may be added to or used in conjunction with the microbial culture and / or growth byproduct composition, including but not limited to polyoxyethylene alkyl ethers and propylene glycol.

[0120] Alkali-silica reaction inhibitors may be used in or in combination with microbial cultures and / or growth byproduct compositions. Alkali-silica reaction inhibitors include, but are not limited to, barium salts, lithium nitrate, lithium carbonate, and lithium hydroxide.

[0121] Moisture-proofing admixtures may be used in or in combination with microbial cultures and / or growth byproduct compositions. Some chemicals that can be used to waterproof concrete include, for example, calcium oleate or ammonium stearate soaps, calcium oleate or ammonium oleate soaps, butyl stearate, and various petroleum products. Waterproofing materials are also used on the exterior surface of concrete. Some materials that can be used to waterproof concrete include rubberized asphalt coatings, bentonite, rubber coatings, and polyurethane coatings.

[0122] In certain embodiments, a gas generating admixture may be added to or used in conjunction with the microbial culture and / or the growth byproduct composition. The most common gas generating admixtures include, but are not limited to, aluminum powder, activated carbon, and hydrogen peroxide.

[0123] In certain embodiments, an anti-dispersion admixture may be added to or used in conjunction with the microbial culture and / or the growth byproduct composition. Cellulose and acrylic polymers are exemplary materials that may be used as anti-dispersion admixtures.

[0124] In certain embodiments, foaming admixtures may be added to or used in combination with the microbial culture and / or growth byproduct composition, including, but not limited to, surfactants and hydrolyzed proteins. In certain embodiments, the concrete composition comprises microbial growth byproducts at a concentration of about 0.001% to about 50% (by weight or volume), about 0.01% to about 10%, about 0.05% to about 1%, or about 0.1% to about 0.5%.

[0125] In certain embodiments, the composition for use on a concrete surface contains microbial growth byproducts at a concentration of about 0.001% to about 50% (by weight or volume of the composition), about 0.01% to about 10%, about 0.01% to about 1%, or about 0.1% to about 0.5%.

[0126] In certain embodiments, the microbial cells may be present in a concentration of, for example, at least 1×10 4 , 1×10 5 , 1×10 6 , 1×10 7 , 1×10 8 , 1×10 9 , 1×10 10 or 1×10 11 or more cells are present.

[0127] Advantageously, according to the present invention, the microbial culture and / or growth byproducts may include a liquid culture medium in which the microorganisms grow. The product may be at least 1 wt%, 5 wt%, 10 wt%, 25 wt%, 50 wt%, 75 wt% or 100 wt% of the liquid culture medium. The amount of biomass in the product may be any value of 0 wt%-100 wt%, including all percentages therebetween.

[0128] Optionally, the product can be stored before use. Shorter storage times are preferred. Thus, the storage time may be less than 60 days, 45 days, 30 days, 20 days, 15 days, 10 days, 7 days, 5 days, 3 days, 2 days, 1 day or 12 hours. In a preferred embodiment, if living cells are present in the product, the product is stored at low temperatures, such as below 20° C., 15° C., 10° C. or 5° C. On the other hand, the biosurfactant composition can generally be stored at ambient temperature.

[0129] Any composition or method provided herein can be combined with one or more of any other compositions and methods provided herein.

[0130] Other features and advantages of the invention will become apparent from the following description of the preferred embodiments, and from the claims.All references cited herein are incorporated herein by reference.

[0131] Use of microorganisms and their growth byproducts in concrete

[0132] The use of concrete comprising the composition according to the present invention will result in a number of advantages. When used in concrete mixtures, the microbial culture and / or growth byproducts can be added during the initial mixing of the various ingredients of the concrete, including water, cement, aggregates and optional admixtures.

[0133] Both acidic and lactone type sophorolipids can be used. The mixture between these two types of sophorolipids can be adjusted according to the desired characteristics of each product use. The ratio between sophorolipids and other ingredients including yeast cells can be adjusted according to each use. Sophorolipids can be purified from organisms that produce chemicals and / or growth media. Alternatively, sophorolipids can be used in an unpurified form. The organisms used to produce biosurfactants can be removed from the composition, while other components of the growth medium and other by-products produced by the organisms can remain in the mixture to be added to the concrete composition.

[0134] As used herein, "using" a composition or product means contacting it with a target, location, or material so that the composition or product can have an effect on the target, location, or material. The effect can be, for example, microbial growth and / or the action of biosurfactants or other growth byproducts. For example, a microbial-based composition or product can be added to a concrete mix in liquid or dry form by pouring, spraying, mixing, spreading, and / or casting at a concrete mixing plant and / or at a site of use of the concrete product.

[0135] When the microbial culture and / or growth byproduct composition is applied to concrete during or after curing, it can be sprayed using, for example, a spray bottle or a pressurized spray device. The composition can also be applied to the surface using a cloth or a brush, spread or brushed onto the surface. In addition, the composition can be applied to the surface by dipping, soaking or immersing the surface into a container containing the composition.

[0136] In one embodiment, the material and / or surface can be soaked in the composition thereon. For example, the soaking time can be at least 5 seconds, 30 seconds, 1 minute, 30 minutes, 60 minutes, 12 hours, 24 hours, 36 hours, 48 ​​hours or 72 hours or longer, as desired.

[0137] In certain embodiments, microbial cultures and / or growth byproducts may improve concrete by increasing the strength of concrete, reducing the amount of water used in concrete compositions, increasing the workability of concrete, reducing the porosity of concrete, changing the speed at which concrete sets, and / or increasing the concrete's resistance to biotic and abiotic stresses.

[0138] Concrete composition

[0139] Microbial cultures and / or growth byproduct compositions can be used in common types of concrete / cement for construction, such as Portland cement. In addition, microbial cultures and / or growth byproduct compositions can be used in asphalt concrete commonly used in roads, parking lots, airports, and dams. Permeable concrete, nanoconcrete, microbial concrete, and polymer concrete can all use microbial cultures and / or growth byproducts as a mixture component.

[0140] Additionally, biosurfactants can be used in concrete coatings. Typically, concrete is coated to seal against penetration of water, salts, and / or air, thereby preventing surface damage, corrosion, and contamination. In certain embodiments, the microbial culture and / or growth byproducts simply coat the surface, while in other embodiments, the microbial culture and / or growth byproducts can penetrate into the concrete and remain on the surface.

[0141] Increase the strength of concrete

[0142] The strength of concrete is generally inversely proportional to the amount of water in the concrete mix. For every 1% reduction in the water concentration in concrete, the strength of the concrete increases by approximately 5%. However, in some cases, reducing the water content can reduce the workability of the concrete and shorten the curing time.

[0143] According to the present invention, the addition of microbial cultures and / or growth byproducts, preferably sophorolipids and Wickhamella anomala, can increase the workability of concrete without compromising strength. Plasticizers and superplasticizers are currently used in concrete mixes to maintain the workability of concrete while increasing concrete strength.

[0144] The strength of concrete can be increased by a combination of methods or by a single method. These methods include, but are not limited to, reducing the amount of water in the concrete composition and reducing the porosity of the concrete. When water evaporates from the concrete during the setting process, voids exist in the concrete; therefore, the smaller the void volume, the stronger the concrete. These resulting voids determine the porosity of the concrete. Using microbial cultures and / or growth byproducts to reduce the amount of water in the initial concrete mix can also reduce the porosity of the concrete. The porosity of the concrete can be determined by the concentration of microbial cultures and / or growth byproducts added to the concrete, and the concrete strength is increased while maintaining workability compared to a concrete mix without the use of microbial cultures and / or growth byproducts.

[0145] In certain embodiments, adding a sophorolipid at a concentration of about 0.01%-0.5% and a yeast cell fermentation waste at a concentration of about 0.1%-0.5% to a concrete mixture can significantly increase the strength of the concrete. The yeast cell fermentation waste can be a product of live yeast cells or dead yeast cells. Therefore, yeast cells can be added at a concentration of about 0.1g / L-100g / L. With the addition of yeast cells, sophorolipids and / or other growth byproducts, water consumption can also be reduced by about 1%-50%, 10%-25%, or 15%-20%. This optional reduction in water can change the curing time of the concrete.

[0146] Changing the curing time of concrete

[0147] By adding microbial cultures and / or growth byproduct compositions to concrete mixes, the curing time can be varied. The water concentration of the concrete determines the length of the curing time (all other variables affecting evaporation being equal). When the water concentration is low, evaporation is faster and the curing time is shorter, while when the water concentration is high, the water evaporates more slowly and the curing time is longer. Advantageously, in certain embodiments, the addition of the compositions of the present invention to the concrete mix can reduce the amount of water required to maintain workability of the concrete. Thus, in certain embodiments, the curing time of the concrete can be shortened.

[0148] Alternatively, in certain embodiments, by adding microbial cultures and / or growth byproduct compositions to concrete mixes, concrete strength can be maintained or even improved while extending curing time. Concrete has a higher plasticity for a longer period of time.

[0149] Improving concrete's resistance to biotic and abiotic stresses

[0150] In certain embodiments, microbial cultures and / or growth byproduct compositions may be added to concrete mixes or applied to the surface of set or curing concrete to increase resistance to environmental stresses, including biotic stresses, such as bacteria and barnacles, and abiotic stresses, including high and low temperatures, freeze-thaw cycles, salt, high or low pressure, wind, precipitation, waves, water, ultraviolet radiation, impurities in the concrete mix or concrete coating, hydrostatic pressure, abrasives (e.g., sandstorms), and movement of concrete, such as when transporting set concrete panels by truck.

[0151] In certain embodiments, by adding microbial cultures and / or growth byproducts, concrete can be made frost resistant. Frost resistance refers to the ability of concrete to maintain its intended use at temperatures below 0°C. To achieve frost resistance, concrete is prevented from cracking by preventing water or moisture from penetrating the concrete, thereby preventing ice from forming within the concrete. In addition, because the water concentration in the composition is lower and / or the amount of air entrained in the concrete is increased, the microbial culture and / or growth byproducts can make the concrete resistant to concrete cracking caused by temperatures below freezing, including below about -30°C, -15°C, or -5°C.

[0152] In certain embodiments, the microbial culture and / or growth byproduct composition extend the life of the concrete by preventing scaling of living or non-living organisms.The present invention can be used to prevent the occurrence of deposits.

[0153] In certain embodiments, the microbial culture and / or growth byproduct composition seals the concrete from water, salt and / or air penetration. The biosurfactant composition can remain on the surface of the concrete, or the composition can penetrate into the concrete. The penetration depth can reach about 0.1%-1%, 0.5%-5%, 1%-10%, 5%-50% or 100% of the total depth of the concrete.

[0154] In certain embodiments, the microbial culture and / or growth byproduct composition can protect the concrete from direct contact with abrasive elements such as waves and wind.

[0155] Evaluating concrete

[0156] In order to identify and evaluate the various properties of a concrete composition or concrete coating, various tests can be performed. These tests can be used to verify whether the concrete meets the structural requirements. Alternatively, these tests can be used to determine the optimal concentration of various admixtures to produce the desired concrete properties.

[0157] Absorption test

[0158] Testing the moisture content in concrete is a common method of determining the water resistance of concrete samples. A common absorption test is available in BS1881-122:2011.

[0159] The British Standard (BS) 1881-122:2011 and British Standard 1881-122:1983 tests measure the amount of water that penetrates a concrete sample when submerged. These tests and test variables are well known to those skilled in the art. In both tests, different items may be required, including: using a balance to weigh the sample; a core drill to cut a soil core of approximately 75 mm in diameter; using a drying oven at a temperature of approximately 105°C; a water tank at least 125 mm deep filled with clean water maintained at a temperature of approximately 20°C; and a dry, sealed container that can be used as a desiccator and has a capacity sufficient to hold three samples to be tested. Each sample is processed according to these art-recognized water permeability tests.

[0160] Rapid chloride permeation test

[0161] The rapid chloride penetration test evaluates the concrete's resistance to chloride penetration by monitoring the amount of charge that passes through a cylindrical specimen. An example of this test can be found in the current standard test method (ASTM) C1202. In this test, an electrical charge is passed through the concrete over a period of six hours. The total charge that passes through the concrete is related to the specimen's resistance to chloride penetration. The lower the level of charge that passes, the higher the resistance. The chloride penetration test is well known to those skilled in the art.

[0162] Water permeability test

[0163] The water permeability test determines the depth to which water will penetrate a concrete sample under hydrostatic pressure. Two examples of water permeability tests can be found in BS (European Standard) EN 12390-8 and DIN 1048 Part 5, which test the depth of water penetration when a concrete sample is subjected to a hydrostatic pressure of 0.5 MPa for 3 days. The concrete sample is cast and cured for 28 days before being placed in a test fixture. The sample holder is open at both ends and one end is subjected to hydrostatic pressure. After 3 days, the sample is removed from the test fixture, split vertically in half, and the maximum depth of water penetration is measured.

[0164] Considering that this test measures the extent to which water penetrates concrete under hydrostatic pressure, it can be used to test concrete used in structures such as basements, tunnels, dams and reservoirs. If the concrete structure will not be subjected to hydrostatic pressure during its service life, this water permeability test can be combined with the absorption test.

[0165] Compressive strength test

[0166] Compressive strength tests check the ability of concrete to perform adequately under static or dynamic loads. Some standard tests include ASTM C31, ASTM C39, ASTM C192, AASHTO T-22, AASHTO T-23, AASHTO T-126, EN1290-1, EN 12390-2, EN 12390-3, EN 12390-4, and EN 12504-1.

[0167] The test can be used to determine the size or structural components required to accommodate the required loads, quality control, approval and acceptability of designed mixes. The test can be carried out at the site where the concrete is used or where it is produced.

[0168] To measure compressive strength, a cylindrical or cubic concrete sample is placed in a compression testing machine, loaded at a constant rate, and the breaking load (the load at which the concrete breaks) is measured. Strength is calculated by dividing the breaking load by the cross-sectional area of ​​the specimen perpendicular to the direction of loading. Strength can be expressed in units of psi, kg / cm 2 Or MPa, and other pressure units.

[0169] The final test result is the average strength of several specimens cast from the same sample and tested at the same curing age, usually specified as full design capacity at 28 days (28-day strength). Three consecutive test results must meet or exceed the specified strength, and no test may be less than a certain percentage of the specified strength.

[0170] In addition to the compression tester, molds, mixers, testing equipment and sample curing equipment are also required.

[0171] Tensile strength test

[0172] Tensile strength testing determines the concrete's ability to resist breaking when tension is applied. Some of the standards for this testing include ASTM C31, ASTM C78, ​​ASTM C192, AASHTO T-23, AASHTO T-97, AASHTO T-126, EN 12390-5, EN 1339, EN 1340, and EN 1521.

[0173] Typically, concrete is not designed to resist direct tensile loads because the tensile strength of concrete may only be 10%-15% of its compressive strength. There are several types of tensile strength tests that can be used to determine the properties of concrete, including direct tension use, flexure, indirect split cylinder testing, and rarely uniaxial tension.

[0174] The flexure test is a common test that involves applying bending loads to an unreinforced concrete prism or beam using symmetrical two-point or three-point loading. The concrete beam will bend under the load, and when the beam fails, the loads at the failure points are used to calculate the tensile strength and / or modulus of rupture. In the split cylinder test, a cylindrical specimen is placed with its long axis horizontally between the platens of a compression machine, and loads are applied incrementally to the sides of the specimen until the cylinder is split along its axis.

[0175] A mold, mixer, test equipment and specimen curing equipment are required. For bending tests, a bending beam frame or bending platen is mounted on a compression machine. For split cylinder tests, a test fixture and wood bars are required to apply the splitting load.

[0176] Density test

[0177] The density of concrete can be determined by different tests, primarily by the buoyancy balance method, in which a known volume is weighed both dry and submerged, and the yield barrel method, in which a sample of fresh concrete is placed in a metal container of known volume. The fresh concrete is compacted into the container, filled to the top, scraped flat on the top, and then weighed. The density is determined by a simple weight / volume calculation. Some of the standard tests that can be used are ASTM C29, ASTM C138, AASHTO T-19, AASHTO T-121, EN 12390-7, and EN 1097-3.

[0178] Moulds, stirrers, buoyancy balances and frames, balances, tamping rods, scraper plates and sample curing equipment are required.

[0179] High-density concrete can be used to cast pre-tensioned reinforcement elements such as tension cables or rebar. Concrete is poured over pre-tensioned cables to provide additional bond strength between the rebar and concrete. Once the concrete hardens, the rebar is released, causing compression of the concrete element. This concrete is used for high-strength elements such as bridges and pre-tensioned slabs. Low-density concrete and aerated concrete are used in ground slabs to improve performance in adverse weather conditions. Concrete density varies depending on the density of its ingredients, including biosurfactants, aggregates and cement, as well as the air content.

[0180] Slump test

[0181] The concrete slump test is used to measure the consistency, workability, and flowability of fresh concrete. Generally, the test shows the ratio of water to cement, with higher water contents showing higher slump values. Slump is an indicator of the compressive strength of hardened concrete. Generally, for a standard weight of concrete, the higher the water content, the lower the strength. However, admixtures including biosurfactants can affect the slump value by changing the ratio of water to cement required for the necessary flowability. Some of the standard tests that can be used are ASTM C143, AASHTO T-119, and EN 12350-2.

[0182] The test is conducted with a standard sized cone-shaped mold called a slump cone or Abrams cone, which is filled with fresh concrete. When the cone is removed, the fresh concrete settles vertically and the slump value is a measure of the vertical settlement or slump from the original height. A metal slump cone, tamper, slump cone base, measuring tape, mold, agitator, test equipment, and sample curing equipment are required.

[0183] High flow concrete is used for pouring concrete within heavy reinforced forms to ensure adequate and uniform distribution of the concrete mix throughout the reinforced concrete member. Therefore, the main purpose of measuring slump is to achieve acceptable workability.

[0184] Air content test

[0185] The air content test is used to determine the amount of air in concrete. The test is usually performed using a pressure method. Concrete is placed in a container of known volume and drained from the top. The method is based on Boyle's law, where the column of air in the void is proportional to the applied pressure. Pressure is applied to the sealed test container by connecting a separate air chamber equipped with a pump. When the valve is closed, the air chamber is pressurized to the calibrated working pressure and the pressure gauge is calibrated. When the valve is opened, the air in the concrete expands into the test chamber and a gauge provides a reading in units of air content. An air delivery meter, scraper, rubber mallet, mixer, test equipment, and specimen curing equipment are required. Some of the standard tests that can be used are ASTM C231, AASHTO T-152, and EN12350-7.

[0186] In areas where concrete is exposed to freeze-thaw cycles, it is often necessary to introduce higher levels of air into the concrete. Water expands when it freezes, which can create internal forces that exceed the bond or tensile strength of the concrete member, causing cracking. Air in the form of very small bubbles provides void spaces within the concrete that act as reservoirs where water can settle and expand, relieving internal pressure during freezing cycles and providing protection to the concrete. Mixing blades are used to evenly distribute the air throughout the concrete mix. Additives are used during the mixing process to stabilize the entrained air bubbles so that they are retained once the concrete hardens.

[0187] Concrete testing hammer

[0188] Test hammers are used to determine the strength of concrete in situ. Concrete test hammers are also called rebound hammers or Schmidt hammers. Test hammers use a spring-driven mass that is released to impact the surface of the concrete sample with a designed amount of energy. The rebound distance after impact is then measured. The hammer is perpendicular to the surface being tested and the rebound varies depending on the hardness of the sample point. The rebound measurement is then converted to compressive strength using conversion tables. Different conversion tables are generated to compensate for instrument orientation. These tables are developed by rebound testing the concrete sample before it is crushed in compression. In addition to the compression tester, molds, mixers, test equipment, and sample curing equipment are also required.

[0189] As mentioned previously, this test is not a standard test for determining compressive strength. However, the number of concrete specimens tested in compression is usually too small to be considered higher than a random test. In many cases, it is also important to determine the compressive strength, modeling, and analysis of aged concrete elements for structural retrofitting.

[0190] Ultrasonic pulse velocity test

[0191] Ultrasonic concrete testers can measure the uniformity of concrete, determine the presence of voids, cracks and other defects, whether the concrete has deteriorated due to ageing or fire frost or chemical attack, measure layer thickness and elastic modulus, and determine concrete strength. To test these parameters, ultrasonic pulses are passed through the concrete and the propagation time is measured.

[0192] This test can be performed on finished concrete to assess the quality of the concrete. When high-speed ultrasonic pulses are observed, the concrete is uniform and homogeneous throughout, in terms of density or the presence of defects. Problems both on the surface and underground can be detected in this way.

[0193] Example

[0194] The present invention and its many advantages can be better understood through the following examples. The following examples illustrate some methods, applications, embodiments and variations of the present invention. The examples should not be considered as limitations of the present invention.

[0195] Example 1 – Increasing the compressive strength of concrete using sophorolipids and Wickham's yeast

[0196] The combination of biosurfactants and yeast cultures improves the strength of concrete. The strength of the samples was evaluated using a compressive strength test. Each sample contained Portland cement, sand and water. Some samples also contained anomalous Wickham yeast and lactones or acid sophorolipids.

[0197] Under the following experimental conditions, the yeast Wickham's yeast and sophorolipids were added to a mixture containing water and In conventional concrete mixes, each kg Add 150ml water, without any admixtures:

[0198] 1. Control: 0g / L abnormal Wickham yeast and 0ml / l sophorolipids

[0199] 2. Group 2: 10 g / L of abnormal Wickham yeast and 0.05 ml / L of lactone-type sophorolipids

[0200] 3. Group 3: 10 g / L abnormal Wickham yeast and 0.1 ml / L lactone-type sophorolipid

[0201] 4. Group 4: 10 g / L abnormal Wickham yeast and 0.3 ml / L acidic sophorolipids

[0202] 5. Group 5: 10 g / L abnormal Wickham yeast and 0.1 ml / L acidic sophorolipid

[0203] 6. Group 6: 10 g / L of abnormal Wickham yeast and 0.05 ml / L of acidic sophorolipids

[0204] The concrete mixes with or without yeast and biosurfactant were poured into cylinders of 50 mm diameter and 100 mm height. The concrete samples were dried at room temperature (22°C) for 27 days.

[0205] To determine the compressive strength of each concrete sample, a TORBAL FB high-resolution precision dynamometer was used. Each concrete sample cylinder was placed vertically on a stand where the gage was leveled and tared. Pressure was applied until the cylinder broke completely and no further force could be measured. The maximum force developed is recorded in Table 1. Each test condition was repeated three times. For each test condition, the average maximum pressure developed was determined.

[0206]

[0207]

[0208] Based on the results shown in Table 1, the compressive strength of the concrete mix containing 10 g / l of Wickham's yeast and 0.05 ml / l of the lactone sophorolipid was 21.4% higher than the control concrete containing no yeast cells or sophorolipid.

Claims

1. A composition for improving concrete, said composition essentially consisting of 10 g / L of an inactive Abnormal Wickham Yeast The method is composed of a yeast culture of 0.05 ml / l of a lactone-type sophorolipid biosurfactant, a liquid culture medium for culturing yeast and / or sophorolipid, and a concrete mixture.

2. The composition according to claim 1, wherein The concrete mixture comprises water, aggregate and cement, wherein the aggregate is selected from one or a combination of the following: sand, stone, blast furnace slag, glass and recycled concrete.

3. The composition according to claim 1, wherein The yeast culture further comprises one or a combination of the following: carbohydrates, polyols, lipids, esters or proteins.

4. The composition according to claim 1, further comprising a concrete admixture.

5. The composition according to claim 4, wherein The concrete admixture comprises at least one of the following: a) accelerators; b) adhesives; c) corrosion inhibitors; d) air-entraining agent; e) Crystallization admixtures; f) Pigments; g) Plasticizers; h) superplasticizers; i) Pumping aids; j) retarders; k) water reducing agent; l) Shrinkage reducing agents; m) hydration control admixtures; n) Alkali-silica reaction inhibitor; o) Moisture-proof admixtures; p) Seepage reducing admixtures; q) Gas generating admixtures; r) Anti-dispersion admixtures; s) foaming admixtures; and t) Workability-enhancing admixtures.

6. A method for improving the compressive strength of concrete, the method comprising adding 10 g / L of an inactive Wickham's yeast The method is composed of a yeast culture of 0.05 ml / l of a lactone-type sophorolipid biosurfactant and a liquid culture medium for culturing yeast and / or sophorolipid.

7. The method according to claim 6, wherein: The concrete contains water, aggregates and cement.

8. The method of claim 7, further comprising a concrete admixture.

9. The method according to claim 8, wherein: The concrete admixture is selected from one or a combination of the following: a) accelerator; b) adhesives; c) corrosion inhibitors; d) air-entraining agent; e) Crystallization admixtures; f) Pigments; g) Plasticizers; h) superplasticizers; i) Pumping aids; j) retarders; k) water reducing agent; l) Shrinkage reducing agents; m) hydration control admixtures; n) Alkali-silica reaction inhibitor; o) Moisture-proof admixtures; p) Seepage reducing admixtures; q) Gas generating admixtures; r) anti-dispersion admixtures; s) foaming admixtures; and t) Workability-enhancing admixtures.

10. The method according to claim 7, wherein: The aggregate is selected from one or a combination of the following: sand, stone, blast furnace slag, glass and recycled concrete.

11. The method according to claim 6, wherein: The yeast culture further comprises one or a combination of the following: carbohydrates, polyols, lipids, esters, and proteins.

12. The method according to claim 6, wherein: Prior to using the concrete, the concrete, the sophorolipid and / or the yeast culture are mixed together.

13. The method according to claim 6, wherein: When the yeast culture and / or the sophorolipid is applied to the concrete, the concrete is curing.

14. The method according to claim 6, wherein: When the yeast culture and / or the sophorolipid is applied to the concrete, the concrete is fully cured.

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