Compositions and methods for flocculation

Abstract

The present invention provides a safe, environmentally friendly composition and an efficient method for flocculation. More specifically, the present invention provides a composition for flocculation derived from microorganisms, which can be used to increase the rate of flocculation and / or the amount of agglomerated particles.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to U.S. Provisional Patent Application No. 63 / 350,213, filed on June 8, 2022, the contents of which are incorporated herein by reference in their entirety. Background Art

[0003] In order to effectively remove surface sediments and reduce pollution associated with various mining and quarrying activities, flocculants are often used. During the mining process, flocculants can be added to adsorb target minerals through electrostatic interactions, hydrogen bonding, or chemical bonding. Currently, starch and benzohydroxamic acid are used as flocculants in iron ore mining to flocculate hematite and other low-grade rare earth elements, minerals, or metals.

[0004] Mining or excavating rock, including quarrying, can also lead to water contamination. During mining or quarrying, rock can be exposed to the atmosphere, including wind and rain, which can cause the minerals to oxidize into toxic chemicals such as cyanide, sulfuric acid, and soluble iron. Water discharged from coal mine tailings and other sources, including iron sulfide and related sulfide minerals, is often referred to as "acid mine drainage" or "acid rock drainage." In addition to sulfuric acid and soluble iron, acid mine drainage can contain high concentrations of heavy metals such as molybdenum, tungsten, chromium, manganese, nickel, arsenic, and vanadium. In addition to wastewater from coal mines, silver, gold, and copper mines can also produce drainage containing toxic dissolved metals. The toxic water can contaminate groundwater and aquifers.

[0005] Several methods have been used to treat mine drainage, including by neutralizing or otherwise treating the drainage. For example, acid mine drainage can be treated with alkaline substances such as calcium carbonate, calcium oxide, calcium hydroxide, sodium hydroxide, sodium carbonate and ammonia. In addition to chemical treatment, acid mine drainage has also been treated using various biological treatment options. Biological mechanisms generally rely on the construction of containment ponds, etc., to subject the acid mine drainage to biological action for a longer period of time.

[0006] Currently, commonly used acid mine treatment methods include chemical precipitation of metals by increasing the pH and / or adding flocculants to the discharge water or wastewater to form metal hydroxides. This is achieved by adding flocculants, such as various forms of lime (such as calcium carbonate and calcium oxide), whereby the pH of the water is increased to cause the precipitation of metal hydroxides. However, some mining areas have low concentrations of iron or other compounds, so in order to flocculate and / or precipitate metal hydroxides, large amounts of lime and other compounds must be added to treat the mine drainage or wastewater. The use of excessive flocculants reduces the efficiency of water treatment and increases costs.

[0007] In addition to their use in mining, flocculants are also used in papermaking, beer brewing, and for treating sewage sludge from municipal wastewater or rainwater. In recent years, the amount of sludge produced has been increasing, and the properties of the sludge have become increasingly worse. However, the amount of sludge that can be treated using conventional flocculants is limited, and the treatment conditions are not satisfactory in terms of the moisture content of the dewatered filter cake, the recovery rate, and the removability of the filter cake from the filter cloth.

[0008] Therefore, there is a need for new and improved flocculation compositions and methods. Summary of the invention

[0009] The present invention generally relates to flocculant compositions and methods of using these compositions. More specifically, the present invention provides environmentally friendly flocculant compositions and methods for flocculation, such as methods for mining, wastewater treatment, papermaking, and beer brewing. In certain embodiments, existing methods can be combined with the compositions and methods of the present invention.

[0010] Advantageously, the compositions and methods of the present invention increase flocculation efficiency and can reduce the use of chemicals required for flocculation (including the use of chemical surfactants). Thus, the present invention can be used to reduce the time required for mining, water treatment (e.g., mining wastewater, municipal wastewater, rainwater, swimming pool water), or production of various products (including, for example, paper and beer).

[0011] In certain embodiments, the present invention provides compositions comprising components derived from microorganisms. In certain embodiments, the compositions comprise microbial biosurfactants. In certain embodiments, the compositions comprise one or more biosurfactants, and optionally other compounds, such as water; chemical surfactants; polymers; pH adjusters; water softeners; ion exchangers; corrosion inhibitors; algaecides; biocides; clarifiers; coagulants; inorganic salts, including aluminum, iron, magnesium and calcium salts; or any combination thereof.

[0012] In certain embodiments, the biosurfactant in the composition is used in a crude form. In addition to the biosurfactant, the crude form may also include fermentation broth of a microorganism that produces the biosurfactant, residual microbial cytoplasm or active or inactive microbial cells, residual nutrients and / or other microbial growth byproducts.

[0013] In some embodiments, the biosurfactant is used after extraction and optional purification from the fermentation broth.

[0014] The biosurfactant according to the present invention may be a glycolipid (such as sophorolipids, rhamnolipids, cellobiose lipids, mannoerythritol lipids and trehalose lipids), a lipopeptide (such as surfactin, iturin, fengycin, actin and lichenin), a flavonoid, a phospholipid (such as cardiolipin), a fatty acid ester compound, a fatty acid ether compound, and / or a high molecular weight polymer (such as lipoprotein, lipopolysaccharide-protein complex and polysaccharide-protein-fatty acid complex).

[0015] In certain specific embodiments, the biosurfactant is a sophorolipid (SLP), including linear SLP, lactone SLP, acetylated SLP, deacetylated SLP, salt-form SLP, esterified SLP derivatives, amino acid-SLP conjugates, and other SLP derivatives or isomers present in nature and / or synthetic production. In a preferred embodiment, the SLP is a linear SLP or a derivatized linear SLP.

[0016] In certain embodiments, the present invention provides a method for flocculation, comprising the steps of contacting the flocculation composition of the present invention with a liquid.

[0017] In certain embodiments, the method enhances or increases the rate of flocculation and / or the amount of flocculated particles, which may have a diameter of less than about 1 mm, about 500 μm, about 100 μm, about 10 μm, about 1 μm, about 100 nm, about 10 nm, or about 1 nm.

[0018] In certain embodiments, the method includes contacting a flocculation composition containing a biosurfactant and optionally other components (e.g., water, chemical surfactants, polymers, pH regulators, water softeners, ion exchangers, corrosion inhibitors, algaecides, biocides, clarifiers, coagulants or inorganic salts) with a liquid. In certain embodiments, the flocculation composition can be applied to the liquid for a period of time, and / or until a specific volume of the composition has been applied. This step can be repeated as many times as desired to achieve the desired amount of agglomerated particles or agglomeration rate.

[0019] In certain embodiments, the flocculating composition of the present invention is effective because it enhances and / or increases the rate or total amount of agglomerated particles in a liquid from a colloidal suspension containing the particles. For example, in certain embodiments, the sophorolipid forms micelles that contain, connect or neutralize the particles, wherein the size of the micelle is less than 500 μm, less than 100 μm, less than 10 μm, less than 1 μm, less than 100 nm, less than 50 nm, less than 25 nm, less than 15 nm or less than 10 nm.

[0020] In certain embodiments, the method of the present invention increases the amount of flocculated particles in the suspension by at least 25%, more preferably at least 50% after one treatment. In certain embodiments, the liquid may be treated multiple times to further increase the amount of flocculated particles.

[0021] Advantageously, in certain embodiments, the flocculation compositions of the present invention are able to effectively agglomerate toxic suspended particles. In addition, the methods of the present invention do not require complex equipment or high energy consumption, and the production of the compositions can be carried out on-site (eg, at a mine site or wastewater treatment facility). DETAILED DESCRIPTION

[0022] The present invention generally relates to the flocculation of suspended particles in liquids. More specifically, the present invention provides environmentally friendly compositions and methods for flocculation of liquids generated in mining areas, wastewater, and water derived from industrial activities. Therefore, the present invention is of great significance for improving the efficiency and effectiveness of flocculation methods. Advantageously, the compositions and methods of the present invention use safe, environmentally friendly compositions to improve the flocculation of particles.

[0023] Selected Definitions

[0024] As used herein, "administering" a composition or product means contacting it with a target or site so that the composition or product has an effect on the target or site. The effect may be caused by the growth of microorganisms and / or the action of biosurfactants or other byproducts of microbial growth, etc.

[0025] As used herein, "biofilm" is a complex collection of microorganisms (e.g., bacteria, yeast, or fungi) in which cells adhere to each other and / or to a surface via an extracellular 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.

[0026] As used herein, an "isolated" or "purified" nucleic acid molecule, polynucleotide, polypeptide, protein or small molecule, or other organic compound (e.g., an organic compound described below) is substantially free of other compounds associated with nature (e.g., cellular material). A purified or isolated polynucleotide (ribonucleic acid (RNA) or deoxyribonucleic acid (DNA)) is free of flanking genes or sequences in its naturally occurring state. A purified or isolated polypeptide is free of amino acids or sequences that flank it in its naturally occurring state. An isolated microbial strain refers to a strain that is removed from its naturally occurring environment. Thus, an isolated strain may exist as a biologically pure culture or as a spore (or other form of strain) associated with a vector.

[0027] In certain embodiments, the purified compound is at least 60% by weight of the compound of interest. Preferably, the preparation is at least 75% by weight of the compound of interest, more preferably at least 90%, and most preferably at least 98%. For example, the purified compound is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 98%, 99% or 100% (w / w) by weight of the desired compound. Purity is measured by any appropriate standard method such as column chromatography, thin layer chromatography or high performance liquid chromatography (HPLC) analytical methods.

[0028] "Metabolite" refers to any substance produced by metabolism or a substance necessary to participate in a specific metabolic process. A metabolite can be an organic compound that is a starting material, intermediate, or end product of metabolism. Examples of metabolites can include, but are not limited to, enzymes, acids, solvents, alcohols, proteins, vitamins, minerals, trace elements, amino acids, biopolymers, and biosurfactants.

[0029] The ranges provided herein are understood to be shorthand expressions of all numerical values ​​within the range. For example, a range of 1 to 20 may be understood to include any number, combination of numbers, or subranges in the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, and 20, as well as all decimal values ​​between the above integers, such as 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, and 1.9. With regard to subranges, "nested subranges" extending from either end of the range are particularly contemplated. For example, a nested subrange of an exemplary range of 1 to 50 may include 1 to 10, 1 to 20, 1 to 30, and 1 to 40 in one direction, or 50 to 40, 50 to 30, 50 to 20, and 50 to 10 in another direction.

[0030] As used herein, "decrease" refers to a negative change and "increase" refers to a positive change, wherein the negative change or the positive change is at least 0.001%, 0.01%, 0.1%, 0.5%, 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100%.

[0031] 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. "Biosurfactants" are surface-active substances produced by living cells and / or using natural extraction substrates.

[0032] Biosurfactants are a group of structurally diverse surface-active substances that consist of two parts: a polar (hydrophilic) portion and a nonpolar (hydrophobic) group. Due to their amphiphilic structure, biosurfactants can increase the surface area of ​​hydrophobic insoluble substances, improve the water bioavailability of these substances, and change the properties of bacterial cell surfaces, etc. Biosurfactants can also reduce the interfacial tension between water and oil, thereby reducing the hydrostatic pressure required to move the entrained liquid to overcome the capillary effect. Biosurfactants accumulate at the interface, thereby reducing the interfacial tension and causing the formation of agglomerated micellar structures in the solution. The formation of micelles provides a physical mechanism for moving oil in the aqueous phase, etc.

[0033] The ability of biosurfactants to reduce surface tension also allows them to be used as antibacterial, antifungal and hemolytic agents to control harmful organisms and / or microbial growth, among other things.

[0034] Typically, the hydrophilic group of a biosurfactant is a sugar (e.g., a monosaccharide, disaccharide, or polysaccharide) or a peptide, while the hydrophobic group is usually a fatty acid. Thus, there are countless variations of biosurfactant molecules, depending on factors such as the type of sugar, the number of sugars, the size of the peptide, the amino acids contained in the peptide, the length of the fatty acid, the degree of saturation of the fatty acid, additional acetylation, additional functional groups, esterification, polarity, and the charge of the molecule.

[0035] These changes have resulted in the formation of a group of molecules including a variety of categories, including, for example, glycolipids (such as sophorolipids, rhamnolipids, cellobiose lipids, mannoerythritol lipids and trehalose lipids, etc.), lipopeptides (such as surfactin, iturin, fengycin, sedative and / or lichenin, etc.), flavonoids, phospholipids (such as cardiolipin), fatty acid ester compounds and high molecular weight polymers (such as lipoproteins, lipopolysaccharide-protein complexes and polysaccharide-protein-fatty acid complexes, etc.). Each biosurfactant in each category may further include subtypes with further modified structures.

[0036] Like chemical surfactants, each biosurfactant molecule has an HLB value that is determined by its structure; however, unlike the production of chemical surfactants, which produces a single molecule with a single HLB value or range, a biosurfactant production cycle typically produces a mixture of biosurfactant molecules (e.g., subtypes and their isomers).

[0037] The phrases "biosurfactant" and "biosurfactant molecule" include all forms, analogs, homologs, isomers and natural and / or artificial modifications of any biosurfactant class (e.g., glycolipids) and / or its subtypes (e.g., sophorolipids).

[0038] As used herein, the term "sophorolipid", "sophorolipid molecule", "SLP" or "SLP molecule" includes all forms of SLP molecules and isomers thereof, including, for example, acidic (straight chain) SLP (ASL) and lactone SLP (LSL). The term "sophorolipid", "sophorolipid molecule", "SLP" or "SLP molecule" further includes monoacetylated SLP, diacetylated SLP, esterified SLP, SLP with different hydrophobic chain lengths, cationic and / or anionic SLPs linked to fatty acid-amino acid complexes, esterified SLPs, SLP metal complexes, SLP salt derivatives (e.g., sodium salts of straight chain SLPs) and other SLPs (including those described and / or not described in the present disclosure).

[0039] In a preferred embodiment, the SLP molecule according to the present invention is represented by the general formula (1) and / or the general formula (2), and comprises fatty acids with different chain lengths (R 3 ) and in some cases, in R 1 and / or R 2 Acetylation or protonation occurs at the site.

[0040]

[0041] In the general formula (1) or (2), R 0 It can be a hydrogen atom or a methyl group. 1 and R 2 are each independently a hydrogen atom or an acetyl group. 3 It is a saturated aliphatic hydrocarbon chain, or an unsaturated aliphatic hydrocarbon chain having at least one double bond, and may have one or more substituents.

[0042] Examples of the substituent include a halogen atom, a hydroxyl group, a lower (C1-C6) alkyl group, a halogenated lower (C1-C6) alkyl group, a hydroxy lower (C1-C6) alkyl group, a halogenated lower (C1-C6) alkoxy group, and the like. 3 Typically has 11 to 20 carbon atoms. In certain embodiments of the invention, R 3 It has 18 carbon atoms.

[0043] SLP is usually produced by yeast, such as Starmerella spp. yeasts and / or Candida, such as Starmerella (Candida) bombicola, Candida apicola, Candida batista, Candida floricola, Candida riodocensis, Candida stellate and / or Candida kuoi, etc. SLP has environmental compatibility, high biodegradability, low toxicity, high selectivity and specific activity under a wide range of temperature, pH and salinity conditions. In addition, in some embodiments, SLP has an advantage due to its smaller micelle size, which helps to promote the movement of micelles and compounds enclosed therein through nanoscale pores and spaces. In certain embodiments, the micelle size of the SLP is less than 100 nm, less than 50 nm, less than 20 nm, less than 15 nm, less than 10 nm, or less than 5 nm.

[0044] As used herein, "flocculation" refers to the process by which colloidal (ie, suspended) particles form clusters of larger sized particles by agglomerating in the form of flocs or flakes.

[0045] As used herein, "ore beneficiation" refers to the process of removing waste material from products of interest (eg, elements, compounds, minerals).

[0046] As used herein, "ore" refers to a naturally occurring solid material from which valuable substances, minerals and / or metals can be extracted. Ores are typically mined from a deposit consisting of ore minerals containing valuable substances. "Gangue" minerals refer to minerals present in a deposit but without valuable substances. Examples of deposits include hydrothermal deposits, magmatic deposits, laterite deposits, volcanogenic deposits, metamorphic alteration deposits, carbonate alkaline igneous rock-related deposits, placer deposits, residual ore deposits, sedimentary deposits, sedimentary hydrothermal deposits, and crater-related deposits. However, ore as defined herein may also include concentrates or tailings.

[0047] As used herein, "leaching" refers to the process of extracting metals from ores by aqueous solution, including ammonia leaching, alkaline leaching, acid leaching, cyanidation (i.e., cyanide leaching) or thiosulfate leaching, etc. As used herein, "cyanidation" refers to the process of converting gold in an ore into a water-soluble coordination complex using an aqueous cyanide solution (including sodium cyanide, potassium cyanide or calcium cyanide).

[0048] As used herein, "colloid" or "colloidal particles" refers to a mixture of one insoluble substance dispersed or suspended in another substance. The insoluble substance is usually dispersed in a liquid.

[0049] 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 "consisting of" excludes any element, step, or ingredient not specified in the claim. The transitional phrase "consisting essentially of" limits the scope of the claim to the specified materials or steps "and those that do not materially affect the basic, novel characteristic." Use of the term "comprising" refers to other embodiments consisting of "consisting of" or "consisting essentially of" the referenced components.

[0050] Unless otherwise specified or obvious from the context, the term "or" as used herein should be understood to be inclusive. Unless otherwise specified or obvious from the context, as used herein, the terms "a", "and" and "the" are understood to be singular or plural.

[0051] Unless otherwise specified or apparent from the context, as used herein, the term "about" is to be understood as being within the normal tolerance range in the art, such as within 2 standard deviations of the mean. "About" may be understood as being within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05%, or 0.01% of the specified value. Unless the context clearly indicates otherwise, all numerical values ​​provided herein are modified by the term "about".

[0052] The recitation of a list of chemical groups in any definition of a variable herein includes definitions of the variable as any single group or combination of the listed groups. The recitation of an embodiment for a variable or aspect herein includes the embodiment as any single embodiment or in combination with any other embodiment or portion thereof.

[0053] All references cited herein are hereby incorporated by reference in their entirety.

[0054] Flocculation composition

[0055] In certain embodiments, the present invention provides compositions comprising components derived from microorganisms. In certain embodiments, the compositions comprise microbial biosurfactants. In certain embodiments, the compositions comprise one or more biosurfactants, and optionally other compounds, such as water; chemical surfactants; polymers; pH adjusters; water softeners; ion exchangers; corrosion inhibitors; algaecides; biocides; clarifiers; coagulants; inorganic salts, including aluminum, iron, magnesium and calcium salts; or any combination thereof.

[0056] In certain embodiments, the chemical surfactant of the flocculating composition is a detergent, wetting agent, emulsifier, foaming agent, and / or dispersant.

[0057] In certain embodiments, the polymer may comprise a natural or synthetic polymer, a water-soluble polymer, a cationic polymer, an anionic polymer or a nonionic polymer. The polymer may be an anionic polyacrylamide, a modified polyacrylamide, a nonionic polyacrylamide, starch, guar gum, Moringa seed extract, Strychnos nux vomica seed extract, gelatin (e.g., isinglass), alginate (e.g., sodium alginate) or chitosan, etc.

[0058] In certain embodiments, the pH adjusting agent can be any compound that upon addition of the compound can produce a pH in the liquid of about 4 to about 10, about 5 to about 9, about 6 to about 8, about 6 to about 7.5, about 6.5 to about 7.5, or about 7. In certain embodiments, the compound is carbon dioxide, sulfuric acid, or sodium hydroxide.

[0059] In certain embodiments, the water softener is ammonia, borax, calcium hydroxide (hydrated lime), trisodium phosphate, or sodium carbonate (soda ash).

[0060] In certain embodiments, the corrosion inhibitor is a chromate, a nitrate, a molybdate, a tungstate, or the like.

[0061] In certain embodiments, the biocide is an herbicide, an insecticide, an algaecide, a molluscicide, a miticide, a piscicide, a rodenticide, a slimicide, a fungicide, an antibiotic, an antibacterial, an antiviral, an antifungal, an antiprotozoal, or an antiparasitic compound.

[0062] In certain embodiments, aluminum salts may include, for example, aluminum sulfate, aluminum chloride, sodium aluminate, alum (i.e., hydrated double sulfate of aluminum), aluminum chlorohydrate, polyaluminum chloride, polyaluminum chlorosulfate, polyaluminum chlorosilicate, and polyaluminum chloride with an organic polymer; iron salts may include ferric sulfate, ferrous sulfate, ferric chloride, ferric sulfate chloride, polyferric sulfate, and iron salts with polymers; magnesium salts may be magnesium carbonate or magnesium sulfate. In certain embodiments, the salt may be a polymerized aluminum-iron blend. In certain embodiments, the coagulant may be lime (e.g., calcium carbonate, calcium oxide) or slaked lime (i.e., calcium hydroxide).

[0063] In certain embodiments, the flocculation composition comprises a microbial-based product including a biosurfactant used in a crude form. In addition to the biosurfactant, the crude form may also include a fermentation broth of a microorganism that is cultured to produce the biosurfactant, residual microbial cytoplasm or active or inactive microbial cells, residual nutrients and / or other microbial growth byproducts. The product may be at least 1%, 5%, 10%, 25%, 50%, 75% or 100% of the fermentation broth by weight. The biomass in the product may be 0% to 100% (including all percentages between 0% and 100%) by weight.

[0064] In some embodiments, the biosurfactant is used after extraction and optional purification from the fermentation broth.

[0065] The biosurfactant according to the present invention may be a glycolipid (such as sophorolipids, rhamnolipids, cellobiose lipids, mannoerythritol lipids and trehalose lipids), a lipopeptide (such as surfactin, iturin, fengycin, actin and lichenin), a flavonoid, a phospholipid (such as cardiolipin), a fatty acid ester compound, a fatty acid ether compound, and / or a high molecular weight polymer (such as lipoprotein, lipopolysaccharide-protein complex and polysaccharide-protein-fatty acid complex).

[0066] In certain specific embodiments, the biosurfactant is a sophorolipid (SLP), including linear SLP, lactone SLP, acetylated SLP, deacetylated SLP, salt-form SLP, esterified SLP derivatives, amino acid-SLP conjugates, and other SLP derivatives or isomers present in nature and / or synthetic production. In a preferred embodiment, the SLP is a linear SLP or a derivatized linear SLP. In certain embodiments, the composition of the present invention may comprise a lactone SLP and a linear SLP, wherein at least about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90% or about 95% of the SLP comprises a linear form, and the remainder comprises a lactone form.

[0067] In some embodiments, the biosurfactant may be included in the composition in a ratio of 0.01% to 99.9%, 0.1% to 90%, 0.5% to 80%, 0.75% to 70%, 1.0% to 50%, 1.5% to 25%, or 2.0% to 15% by weight relative to the total amount of the flocculation composition.

[0068] In another embodiment, the purified biosurfactant may be added with an acceptable carrier, as the biosurfactant may be present in a concentration of 0.001% to 50% (v / v), preferably 0.01% to 20% (v / v), more preferably 0.02% to 5% (v / v).

[0069] In some embodiments, the biosurfactant can be included in the composition at a concentration of 0.01 ppm to 100,000 ppm, 0.05 ppm to 10,000 ppm, 0.1 ppm to 1,000 ppm, 0.5 ppm to 750 ppm, 1.0 ppm to 500 ppm, 2.0 ppm to 250 ppm, or 3.0 ppm to 100 ppm, etc., relative to the amount of liquid being treated.

[0070] In certain embodiments, the chemical surfactant of the flocculation composition is a detergent, a wetting agent, an emulsifier, a foaming agent and / or a dispersant. In some embodiments, the chemical surfactant can be included in the composition in a proportion of 0.01% to 99.9%, 0.1% to 90%, 0.5% to 80%, 0.75% to 70%, 1.0% to 50%, 1.5% to 25% or 2.0% to 15% by weight relative to the total amount of the flocculation composition.

[0071] The flocculation composition may further include other additives, such as carriers, other microbial-based compositions, additional biosurfactants, enzymes, catalysts, solvents, salts, buffers, chelating agents, acids, emulsifiers, lubricants, solubility regulators, preservatives, stabilizers, UV inhibitors, viscosity regulators, preservatives, tracers, antimicrobials and other microorganisms and other ingredients specific to the intended use.

[0072] In certain embodiments, the chelating agent may be, but is not limited to, ethylenediaminetetraacetic acid (EDTA), nitrilotriacetic acid (NTA), phosphonates, succinic acid (DMSA), diethylenetriaminepentaacetate (DTPA), N-acetylcysteine, N-hydroxyethylethylenediaminetriacetic acid (HEDTA), organic acids with multiple coordinating groups (e.g., erythromycin), STPP (sodium tripolyphosphate, Na 5 P 3 O 10 ), trisodium phosphate (TSP), water, carbohydrates, organic acids with more than one coordinating group (e.g., citric acid), lipids, steroids, amino acids or related compounds of the above (e.g., glutathione), peptides, phosphates, nucleotides, tetrapyrroles, ferroamines, ionophores, or phenols, sodium citrate, sodium gluconate, ethylenediamine disuccinic acid (EDDS), iminodisuccinic acid (IDS), L-glutamic acid diacetic acid (GLDA), GLDA Na 4, methylglycine diacetic acid (MGDA), polyaspartic acid (PASA), hemoglobin, chlorophyll, lipophilic β-diketone and (14,16)-heteronedione, ethylenediamine-N,N'-diglutaric acid (EDDG), ethylenediamine-N,N'-dimalonic acid (EDDM), 3-hydroxy-2,2-iminodisuccinic acid (HIDS), 2-hydroxyethyliminodiacetic acid (HEIDA), pyridine-2,6-dicarboxylic acid (PDA), trimethylglycine (TMG), titanium reagent, or any combination thereof.

[0073] Flocculation method

[0074] In certain embodiments, the present invention provides a method for flocculating liquids from various sources including mining, quarrying, wastewater, agricultural, and industrial sources, among others.

[0075] In certain embodiments, the present invention provides a method for flocculating tailings of a mine. The method comprises adding a composition of the present invention to the tailings and removing the agglomerated tailings particles. By flocculating the tailings, the suspended particles can reach a higher concentration when agglomerated. In a preferred embodiment, the tailings are low-grade tailings, wherein the tailings account for less than about 50%, about 40%, about 35%, about 30% or about 25% of the product of interest (e.g., the metal, mineral, compound or element mined), and the remainder comprises gangue.

[0076] In certain embodiments, the mining site may be a coal mine, an iron ore mine, a copper mine, a copper-nickel mine, a tin mine, a nickel mine, a gold mine, a silver mine, a molybdenum mine, an aluminum mine (e.g., a bauxite mine, a kyanite mine), a lead-zinc mine, a tungsten mine, or a zinc mine. The mine may be an underground mine, an open-pit mine, a placer mine, or an in-situ mine. In certain embodiments, various toxic compounds may originate from mining activities. In certain embodiments, according to the method of the present invention, a method for removing the toxic compounds is provided by contacting the flocculated compounds with various water flows, pipes, pumps, water storage areas, or other aquatic environments. Toxic compounds may include cyanide, sulfur-containing minerals, soluble iron, and heavy metals (such as molybdenum, tungsten, chromium, manganese, nickel, arsenic, and vanadium, etc.).

[0077] In certain embodiments, the microorganism-containing and / or biosurfactant-containing compositions can enhance the agglomeration of particles or between particles and surfaces, such as agglomerating toxic byproducts of coal mining with each other.

[0078] In certain embodiments, the composition containing microorganisms and / or the composition containing biosurfactants can form a layer of agglomerated particles around and / or between particles suspended in a liquid. In certain embodiments, the composition containing microorganisms and / or the composition containing biosurfactants can neutralize charged particles.

[0079] In certain embodiments, the composition can reduce the effects of wind, rain or physical movement of rock, and is preferably used in a method of reducing toxic compounds in drainage. Examples of physical movement include tilling, mining, tunneling, digging, hauling, dumping, crushing, blasting, separating, pelletizing, cutting, casting, quarrying, dredging, fracturing, or any combination thereof.

[0080] These compositions can be applied to liquids at certain temperatures and in aquatic environments, such as streams, rivers, waterways, oceans, seas, lakes, ponds, runoff areas, containment ponds, or wastewater treatment / storage tanks.

[0081] The flocculating composition may be applied to a liquid and optionally mixed by adding, pouring or combining.

[0082] In certain embodiments, the flocculating composition may be in contact with the liquid for a period of about 1 second to about 1 year, about 1 minute to about 1 year, about 1 minute to about 6 months, about 1 minute to about 1 month, about 1 minute to about 1 week, about 1 minute to about 48 hours, about 30 minutes to 40 hours, or preferably about 12 hours to 24 hours. In certain embodiments, the method includes applying the liquid form of the flocculating composition to the liquid during the period of time during which the liquid containing suspended particles is produced, or until the amount of suspended particles is reduced to an amount determined to be satisfactory or safe, which can be easily determined by a person skilled in the art. The amount of suspended particles can be considered to be an acceptable and / or safe amount depending on the circumstances. For example, in a mining site where personal protective equipment is required, the amount of suspended particles may be higher than the amount of suspended particles in the treated water of a municipal wastewater treatment facility, which injects the treated water into an aquifer or other waterway.

[0083] In certain embodiments, the amount of flocculating composition applied is about 0.00001 to 15%, about 0.00001 to 10%, about 0.0001 to 5%, about 0.001 to 3%, about 0.01%, or about 1 vol% based on the amount of liquid treated.

[0084] In certain embodiments, the method of the present invention increases the amount of flocculated particles in the suspension by at least 25%, more preferably at least 50% after one treatment. In certain embodiments, the liquid may be treated multiple times to further increase the amount of flocculated particles.

[0085] In certain embodiments, the flocculation compositions according to the present invention are effective due to the amphiphilic mediated adhesion of the suspended particles. In some embodiments, the sophorolipids or other biosurfactants are used as carriers to promote the agglomeration of particulate matter and / or the adhesion of particulate matter to surfaces and / or objects. For example, in some embodiments, the sophorolipids will form micelles containing particles, wherein the size of the micelles is less than 1 mm, 100 μm, 50 μm, 20 μm, 10 μm, 1 μm, 100 nm, less than 50 nm, less than 25 nm, less than 15 nm or less than 10 nm. The small size and amphiphilic nature of the micelles enhance the adhesion of the particles, so that larger particle agglomerations can occur.

[0086] In certain embodiments, the flocculating composition can be used in methods of processing ore, ore slurry, or other products obtained by mining. In certain embodiments, the flocculating composition can be used for dewatering before grinding, concentrate dewatering, tailings dewatering, tailings filling, middlings dewatering, or any combination thereof.

[0087] In certain embodiments, the flocculation composition can be used in the mineral processing process, especially in low-grade ores containing low concentrations of elements or other products of interest (e.g., gold or silver). In order to extract the elements or compounds of interest, it may be necessary to crush and grind the ore and pre-concentrate or separate the elements or products of interest from the ore by flotation or gravity sorting. In certain embodiments, by adding the flocculation composition in the mineral processing process, the sedimentation rate of the concentrate can be accelerated and the dewatering efficiency can be improved.

[0088] In certain embodiments, the flocculation composition can be used in leaching methods, such as cyanidation gold extraction. The process of extraction by leaching includes leaching (e.g., leaching cyanide), washing and filtering of the leached slurry, extracting metals from the leachate or leached slurry, and smelting of the finished product. In certain embodiments, the flocculation composition can be used in methods for washing and filtering leached slurry, wherein the flocculation composition increases the agglomeration rate of impurities. In certain embodiments, the flocculation composition can agglomerate heavy metals and metalloids (including As, Cd, Co, Cu, Hg, Mn, Ni, U and Zn) in mining tailings, etc., which may bring significant potential ecological risks and human health risks associated with exposure to metals and metalloids in contaminated soils around mining tailings storage sites.

[0089] In certain embodiments, the flocculation composition can be used in methods for treating industrial sewage and sewage containing water-soluble organic matter. The purification process uses activated sludge treatment to remove soluble organic matter. In certain embodiments, the activated sludge treatment uses the growth of microorganisms for treatment, so the dewatering treatment of sludge is generally used to reduce the volume of sludge. In certain embodiments, sludge dewatering can use flocculants. In certain embodiments, the flocculation composition of the present invention can replace synthetic polymer flocculants or be used in combination with polymer flocculants to agglomerate and dehydrate the sludge. Cationic flocculants can neutralize the negative charge on the surface of colloidal particles in the sludge and can bridge between particles to form large and strong floccules for easy dewatering. Sludge containing floccules can be dehydrated by sludge dewatering equipment, separated into solids called sludge cakes, and disposed of by landfill, incineration or composting. In certain embodiments, the flocculation method can reduce the amount of water in the sludge by at least about 50%, about 60%, about 70% or about 80%.

[0090] In certain embodiments, the flocculating compositions can be used in various industrial processes such as the manufacture or processing of food, beverages, oil sands and paper. In product manufacturing processes, the flocculating compositions are primarily used to treat wastewater (i.e., to agglomerate sludge), where suspended particles (e.g., sludge, phosphate-containing particles, and residual oil sands in oil sand tailings) are agglomerated and then removed from the liquid before discharging or storing the water. In certain embodiments, the flocculating compositions can also be used in production processes (e.g., in the fermentation step of beer brewing), where yeast cells and lysed yeast cells can be agglomerated and then removed from the liquid.

[0091] Advantageously, in certain embodiments, the flocculation compositions according to the present invention provide enhanced or improved particle agglomeration efficiency while having limited negative impact on the environment. In addition, the methods of the present invention do not require complex equipment or high energy consumption, and the production of the flocculation compositions can be carried out on site (including mines or industrial sites, etc.). In certain embodiments, the flocculation compositions of the present invention can reduce the use of chemical surfactants, synthetic flocculants, or other potentially harmful chemicals used for flocculation.

[0092] Production of microbial-based products

[0093] In certain embodiments, the present invention provides methods for cultivating microorganisms and generating microbial metabolites and / or other byproducts of microbial growth. The present invention further utilizes culture methods suitable for cultivating microorganisms and generating microbial metabolites on a desired scale. These culture processes include, but are not limited to, liquid culture / fermentation, solid state fermentation (SSF) and modifications, hybrids and / or combinations thereof.

[0094] Microorganisms can be bacteria, yeast and / or fungi, etc. These microorganisms can be natural microorganisms or genetically modified microorganisms. For example, microorganisms can be transformed with specific genes to show specific characteristics. Microorganisms can be mutants of desired strains. As used herein, "mutant" refers to a strain, genetic variant or subtype of a standard bacterium, wherein the mutant has one or more genetic variation forms (e.g., point mutation, missense mutation, nonsense mutation, deletion, duplication, frameshift mutation or amplification mutation) compared to a reference microorganism. The process of preparing mutants is well known in the field of microbiology. For example, UV mutagenesis and nitrosoguanidine are widely used for this purpose.

[0095] In certain embodiments, the microorganism is capable of producing amphiphilic molecules, enzymes, proteins and / or biopolymers. Microbial biosurfactants are particularly produced by a variety of microorganisms, such as bacteria, fungi and yeasts, including the following microbial species, etc.: Agrobacterium spp. (e.g., A. radiobacter); Arthrobacter spp.; Aspergillus spp.; Aureobasidium spp. (e.g., A. pullulans); Azotobacter (e.g., A. vinelandii, A. chroococcum); Azospirillum spp. (e.g., A. brasiliensis); Bacillus spp. (e.g., Bacillus subtilis B.subtili, Bacillus amyloliquefaciens B.amyloliquefaciens, Bacillus pumillus B.cereus, Bacillus licheniformis B.firmus, Bacillus laterosporus B.megaterium); Blakeslea; Candida spp. (e.g., C.albicans, C.rugosa, C.tropicalis, C.lipolytica, C.torulopsis); Clostridium (e.g., C.butyricum, C.tyrobutyricum, C.acetobutylicum, C.beijerinckii); Campylobacter spp.; Cornybacterium spp.; Cryptococcus spp.; Debaryomyces spp. (e.g. D. hansenii); Entomophthora spp.; Flavobacterium spp.; Gordonia spp.; Hansenula spp.; Hanseniaspora spp. (e.g. H. uvarum); Issatchenkia spp.; Kluyveromyces spp.; Meyerozyma spp. (e.g., M. guilliermondii); Mortierella spp.; Mycorrhiza spp.; Mycobacterium spp.; Nocardia spp.; Pichia spp. (e.g., Pichia anomala, Pichia guilliermondii, Pichia occidentalis, Pichia kudriavzevii); Phycomyces spp.; Phythium spp.; Pseudomonas spp. (e.g., Pseudomonas aeruginosa, Pseudomonas chlororaphis, Pseudomonas putida, Pseudomonas florescens, Pseudomonas fragi, Pseudomonas syringae); Pseudozyma spp. (e.g., Pseudomonas aeruginosa, Pseudomonas chlororaphis, Pseudomonas putida, Pseudomonas florescens, Pseudomonas fragi, Pseudomonas syringae); Pseudozyma spp. (e.g., Pseudomonas aphidis); Ralslonia spp. (e.g., Ralslonia eulropha); Rhodococcus spp. (e.g., Rhodococcus erythropolis); Rhodospirillum spp. (e.g., Rhodospirillum rubrum); Rhizobium spp.; Rhizopus spp.; Saccharomyces spp. (e.g., S. cerevisiae, S. boulardiisequela, S. torula); Sphingomonas spp. (e.g., S. paucimobilis); Starmerella spp. (e.g., S. bombicola); Thraustochytrium spp.; Torulopsis spp.; Ustilago spp. (e.g., U. maydis); Wickerhamomyces spp. (e.g., W. anomalus); Williopsis spp.; and / or Zygosaccharomyces spp.(e.g. Z.bailii).

[0096] In a preferred embodiment, the microorganism is Starmerella and / or Candida, such as Starmerella (Candida) bombicola, Candida apicola, Candida batista, Candida floricola, Candida riodocensis, Candida stellate and / or Candidakuoi, etc. In a specific embodiment, the microorganism is Starmerella (Candida) bombicola, such as strain ATCC 22214.

[0097] As used herein, "fermentation" refers to the cultivation or growth of cells under controlled conditions. Growth can be aerobic or anaerobic. In a preferred embodiment, the microorganisms are grown using SSF and / or a modified form thereof.

[0098] In one embodiment, the invention provides materials and methods for producing biomass (e.g., living cellular material), extracellular metabolites (e.g., small molecules and excreted proteins), residual nutrients, and / or intracellular components (e.g., enzymes and other proteins).

[0099] The microbial growth vessel used according to the present invention can be any fermenter or culture reactor for industrial use. In one embodiment, the vessel can have functional controls / sensors or can be connected to functional controls / sensors to measure important factors in the culture process, such as pH, oxygen, pressure, temperature, humidity, microbial density and / or metabolite concentration, etc.

[0100] In another embodiment, the vessel can also monitor the growth of microorganisms in the container (e.g., measuring the number of cells and the stage of growth). Alternatively, daily samples can be taken from the vessel and counted by techniques known in the art (e.g., dilution plate technique). Dilution plate method is a simple technique for estimating the number of organisms in a sample. This technique can also provide an indicator by which different environments or treatments can be compared.

[0101] In one embodiment, the method comprises supplementing the culture with a nitrogen source. The nitrogen source may be, 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 of two or more.

[0102] The method can provide oxygenation for the growing culture. One embodiment utilizes the slow movement of air to remove air low in oxygen content and introduce oxygenated air. In the case of submerged fermentation, the oxygenated air can be ambient air that is replenished daily by a mechanism including an impeller for mechanically agitating the liquid and an air distributor for supplying gas bubbles to the liquid to dissolve oxygen into the liquid.

[0103] 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, rapeseed oil, rice bran oil, olive oil, corn oil, sesame oil and / or linseed oil. These carbon sources may be used alone or in combination of two or more.

[0104] In one embodiment, the growth factors and micronutrients of the microorganisms are included in the culture medium. This is particularly preferred when the growing microorganisms cannot generate all the vitamins they need. Inorganic nutrients may also be included in the culture medium, and the inorganic nutrients include trace elements such as iron, zinc, copper, manganese, molybdenum and / or cobalt. In addition, the source of vitamins, essential amino acids and trace elements can be included in the culture medium in the form of fine powder or coarse powder (for example, corn flour) or in the form of extracts (for example, yeast extract, potato extract, beef extract, soybean extract, banana peel extract, etc.) or in purified form. Amino acids (for example, amino acids that can be used for protein biosynthesis) may also be included.

[0105] In one embodiment, the culture medium may also include inorganic salts. Available inorganic salts may be potassium dihydrogen phosphate, dipotassium hydrogen phosphate, disodium hydrogen phosphate, magnesium sulfate, magnesium chloride, iron sulfate, iron chloride, manganese sulfate, manganese chloride, zinc sulfate, lead chloride, copper sulfate, calcium chloride, sodium chloride, calcium carbonate and / or sodium carbonate. These inorganic salts may be used alone or in a combination of two or more.

[0106] In some embodiments, the culturing method may further include adding additional acid and / or antimicrobial agents to the culture medium before and / or during the culturing process.Antimicrobial agents or antibiotics are used to protect the culture from contamination.

[0107] Additionally, antifoaming agents may be added to prevent the formation and / or accumulation of foam during submerged cultivation.

[0108] The pH of the mixture should be appropriate for the microorganism of interest. Buffers and pH adjusters (e.g., carbonates and phosphates) can be used to stabilize the pH near the preferred value. When metal ions are present in high concentrations, it may be necessary to use a chelating agent in the culture medium.

[0109] The microorganisms can grow in a planktonic form or in a biofilm. If a biofilm is present, the vessel can contain a substrate on which the microorganisms can grow in a biofilm state. The system can also have the ability to apply stimuli (e.g., shear stress) that encourage and / or improve biofilm growth characteristics.

[0110] In one embodiment, the culture method of the microorganism is carried out at about 5° to about 100° C., preferably 15° C. to 60° C., more preferably 25° C. to 50° C. In another embodiment, the culture can be carried out continuously at a constant temperature. In another embodiment, the culture can be subjected to temperature changes.

[0111] In one embodiment, the equipment used in the method and the culture process is a sterile device. Culture equipment (e.g., reactor / vessel) can be separated from a sterilizing device (e.g., autoclave), but connected thereto. Culture equipment can also have a sterilizing device that sterilizes on the spot before starting inoculation. Air can be sterilized by methods known in the art. For example, ambient air can pass through at least one filter before being introduced into the vessel. In other embodiments, culture medium can be pasteurized, or alternatively not heated at all, but low water activity and low pH value can be utilized to control the growth of undesirable bacteria.

[0112] In one embodiment, the present invention further provides a method for producing microbial metabolites by culturing the microbial strain of the present invention under conditions suitable for growth and metabolite production, and optionally purifying the metabolites, the microbial metabolites being biosurfactants, enzymes, proteins, ethanol, lactic acid, β-glucan, peptides, metabolic intermediates, polyunsaturated fatty acids and lipids, etc. The content of the metabolites produced by the method can be at least 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90%, etc.

[0113] Microbial growth byproducts produced by the microorganism of interest can be retained in the microorganism or secreted into the growth medium. The medium can contain compounds that stabilize the activity of the microbial growth byproducts.

[0114] The biomass content of the fermentation medium may be from 5 g / l to 180 g / l or more, or from 10 g / l to 150 g / l, etc.

[0115] The cell concentration can be at least 1×10 6 Up to 1×10 12 , 1×10 7 Up to 1×1011 , 1×10 8 Up to 1×10 10 , or 1×10 9 CFU / ml, etc.

[0116] The methods and apparatus for culturing microorganisms and producing microbial byproducts can be performed in a batch, quasi-continuous process, or a continuous process.

[0117] In one embodiment, all of the microbial culture composition is removed when the culture is complete (eg, when a desired cell density or density of a particular metabolite is reached). In this batch process, a completely new batch is started when the first batch is harvested.

[0118] In another embodiment, only a portion of the fermentation product is removed at any time. In this embodiment, the biomass with living cells, spores, conidia, hyphae and / or mycelium remains in the vessel as an inoculant for a new culture batch. The removed composition can be a cell-free culture medium or contain cells, spores or other reproductive propagules, and / or a combination thereof. A quasi-continuous system is created in this way.

[0119] Advantageously, the method does not require complex equipment or high energy consumption. The microorganisms of interest can be cultured and utilized on-site on a small or large scale, even while still mixed with their culture medium.

[0120] In certain embodiments, the present invention provides a "microorganism-based composition", which means a composition comprising components produced by the growth of microorganisms or other cell cultures. Therefore, a microorganism-based composition may contain the microorganism itself and / or microorganism growth byproducts. The microorganism may be in an asexual state, spore form, hyphae form, any other form of propagule, or a mixture of these. The microorganism may be in the form of a planktonic microorganism or a biofilm, or a mixture of the two. The byproducts of growth may be metabolites, cell membrane components, expressed proteins and / or other cell components, etc. The microorganism may be a complete microorganism or a lysed microorganism. The microorganism may be present in the composition or removed from the composition. The microorganism may be present in the microorganism-based composition together with the fermentation broth in which they are cultured. For example, the cells may be at least 1×10 per milliliter of the composition. 3 , 1×10 4 , 1×10 5 , 1×10 6 , 1×10 7 , 1×10 8 , 1×10 9 , 1×10 10 , 1×10 11 , 1×10 12 , 1×10 13 or more CFUs are present.

[0121] The present invention further provides "microorganism-based products" as products that are applied in practice to achieve the desired effect. Microorganism-based products can be only microorganism-based compositions obtained from microbial culture processes. Alternatively, microorganism-based products can contain other ingredients that have been added. For example, these other ingredients can include nutrients such as stabilizers, acids, buffers, carriers (e.g., water, saline solutions or any other appropriate carriers), nutrients added to support further growth of microorganisms, non-nutritional growth promoters and / or agents that help track microorganisms and / or compositions in their application environment. Microorganism-based products can also include mixtures based on microorganism-based compositions. Microorganism-based products can also include one or more components based on microorganism-based compositions, which have been treated in some way, such as but not limited to filtration, centrifugation, lysis, drying, purification, etc.

[0122] A microbial-based product of the present invention contains only a fermentation medium containing microorganisms and / or microbial metabolites produced by the microorganisms and / or any residual nutrients. The fermented product can be used directly without extraction or purification. If necessary, extraction and purification can be easily achieved using standard extraction and / or purification methods or techniques described in the literature.

[0123] The microorganisms in the microbial-based products can be in active or inactive form, and can also be vegetative cells, reproductive spores, conidia, mycelium, hyphae or any other form of microbial propagation bodies. The microbial-based products can also contain any combination of these forms of microorganisms.

[0124] In one embodiment, different strains of microorganisms are grown separately and then mixed together to produce a microbial-based product. The microorganisms may optionally be blended with the medium in which they were grown and dried prior to mixing.

[0125] The microbial-based products can be used without further stabilization, preservation and storage. Advantageously, the direct use of these microbial-based products maintains high viability of the microorganisms, reduces the possibility of contamination by foreign agents and undesirable microorganisms, and maintains the activity of byproducts of microbial growth.

[0126] After the microbial composition is harvested from the growth vessel, other components may be added when the harvested product is placed in a container or otherwise transported for use. For example, the additives may be buffers, carriers, other microbial compositions produced in the same facility or in a different facility, viscosity modifiers, preservatives, nutrients for microbial growth, surfactants, emulsifiers, lubricants, solubility modifiers, tracking agents, solvents, bactericides, antibiotics, pH adjusters, chelating agents, stabilizers, UV inhibitors, other microorganisms, and other appropriate additives commonly used in such preparations.

[0127] Alternatively, the product may be stored prior to use. The storage time is preferably short. 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 a cool temperature, for example, below 20°C, 15°C, 10°C or 5°C. In another aspect, the biosurfactant composition can generally be stored at ambient temperature.

Claims

1. A flocculation method comprising contacting a flocculation composition comprising a biosurfactant with a liquid containing suspended particles to produce agglomerated particles.

2. The method of claim 1, further comprising removing the agglomerated particles from the liquid. The method according to claim 1 , wherein the agglomerated particles include toxic substances or gangue.

4. The method of claim 3, wherein the toxic substance is cyanide, sulfur-containing minerals, soluble iron, molybdenum, tungsten, chromium, manganese, nickel, arsenic or vanadium.

5. The method according to claim 1, wherein the liquid is waste water or ore slurry.

6. The method according to claim 5, wherein the wastewater is municipal wastewater, rainwater, mining wastewater, quarrying wastewater or industrial wastewater.

7. The method according to claim 5, wherein the wastewater or the ore slurry is from a coal mine, an iron ore mine, a copper mine, a copper-nickel mine, a tin mine, a nickel mine, a gold mine, a silver mine, a molybdenum mine, an aluminum mine, a lead-zinc mine, a tungsten mine or a zinc mine.

8. The method according to claim 7, wherein the aluminum ore is kyanite ore or bauxite.

9. The method of claim 1, wherein the flocculating composition further comprises a chemical surfactant, a polymer, a pH adjuster, a water softener, an ion exchanger, a corrosion inhibitor, a biocide, a clarifier, a coagulant, an inorganic salt, or any combination thereof.

10. The method of claim 1, wherein the flocculating composition is in liquid form, and wherein the contacting step comprises mixing the composition with the liquid for a period of time of about 1 second to about 1 year.

11. The method according to claim 1, wherein the biosurfactant is a sophorolipid and / or a yeast culture comprising a sophorolipid.

12. The method according to claim 11, wherein the yeast culture is of the genus Stamosaccharomyces and / or Candida.

13. The method of claim 11, wherein the yeast is in a vegetative state.

14. The method of claim 11, wherein the yeast is in the form of spores.

15. The method of claim 1, wherein the flocculation comprises agglomerating colloidal particles by one or a combination of: a. Adhere particles to surfaces and / or objects; b. Agglomerate the particles together; c. Settled particles; or d Neutralizing particles.

16. A flocculation composition comprising a sophorolipid and / or a yeast culture comprising a sophorolipid and one or more traditional flocculation components.

17. The composition according to claim 16, wherein the yeast culture is Stamosaccharomyces and / or Candida.

18. The composition of claim 16, wherein the yeast is in a vegetative state.

19. The composition of claim 16, wherein the yeast is in spore form.

20. The composition of claim 16, wherein the traditional flocculation components are selected from one or a combination of the following: a chemical surfactant; b polymer; c pH adjuster; d. Water softener; e ion exchanger; f Corrosion inhibitors; gBiocides; h clarifier; i Coagulant; or jInorganic salts.

21. The composition of claim 20, wherein the inorganic salt is aluminum sulfate, aluminum chloride, sodium aluminate, alum, aluminum chlorohydrate, polyaluminum chloride, polyaluminum chlorosulfate, polyaluminum chlorosilicate, polyaluminum chloride containing organic polymers, ferrous sulfate, ferric chloride, ferric chloride sulfate, polyferric sulfate, iron salt containing polymers, magnesium carbonate, magnesium sulfate, polyaluminum iron blends, lime, or any combination thereof.

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