Classifications

• • C—CHEMISTRY; METALLURGY
  • C22—METALLURGY; FERROUS OR NON-FERROUS ALLOYS; TREATMENT OF ALLOYS OR NON-FERROUS METALS
  • C22B—PRODUCTION AND REFINING OF METALS; PRETREATMENT OF RAW MATERIALS
  • C22B3/00—Extraction of metal compounds from ores or concentrates by wet processes
  • C22B3/18—Extraction of metal compounds from ores or concentrates by wet processes with the aid of microorganisms or enzymes, e.g. bacteria or algae
• • C—CHEMISTRY; METALLURGY
  • C22—METALLURGY; FERROUS OR NON-FERROUS ALLOYS; TREATMENT OF ALLOYS OR NON-FERROUS METALS
  • C22B—PRODUCTION AND REFINING OF METALS; PRETREATMENT OF RAW MATERIALS
  • C22B1/00—Preliminary treatment of ores or scrap
• • C—CHEMISTRY; METALLURGY
  • C22—METALLURGY; FERROUS OR NON-FERROUS ALLOYS; TREATMENT OF ALLOYS OR NON-FERROUS METALS
  • C22B—PRODUCTION AND REFINING OF METALS; PRETREATMENT OF RAW MATERIALS
  • C22B3/00—Extraction of metal compounds from ores or concentrates by wet processes
  • C22B3/20—Treatment or purification of solutions, e.g. obtained by leaching
  • C22B3/26—Treatment or purification of solutions, e.g. obtained by leaching by liquid-liquid extraction using organic compounds
• • C—CHEMISTRY; METALLURGY
  • C22—METALLURGY; FERROUS OR NON-FERROUS ALLOYS; TREATMENT OF ALLOYS OR NON-FERROUS METALS
  • C22B—PRODUCTION AND REFINING OF METALS; PRETREATMENT OF RAW MATERIALS
  • C22B3/00—Extraction of metal compounds from ores or concentrates by wet processes
  • C22B3/20—Treatment or purification of solutions, e.g. obtained by leaching
  • C22B3/26—Treatment or purification of solutions, e.g. obtained by leaching by liquid-liquid extraction using organic compounds
  • C22B3/41—Treatment or purification of solutions, e.g. obtained by leaching by liquid-liquid extraction using organic compounds using a solution of normally solid organic compounds, e.g. dissolved polymers, sugars, or the like
• • C—CHEMISTRY; METALLURGY
  • C22—METALLURGY; FERROUS OR NON-FERROUS ALLOYS; TREATMENT OF ALLOYS OR NON-FERROUS METALS
  • C22B—PRODUCTION AND REFINING OF METALS; PRETREATMENT OF RAW MATERIALS
  • C22B3/00—Extraction of metal compounds from ores or concentrates by wet processes
  • C22B3/20—Treatment or purification of solutions, e.g. obtained by leaching
  • C22B3/44—Treatment or purification of solutions, e.g. obtained by leaching by chemical processes
• • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
  • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
  • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
  • Y02P10/00—Technologies related to metal processing
  • Y02P10/20—Recycling

Definitions

• Low-grade or low-value deposits can be mined from the earth using various mining and quarrying practices.
• solvent extractants can be added to separate certain cations or anions and sequester them from an aqueous phase into an organic phase, thus separating them from other unwanted ionic species.
• cheating agents are used in connection with leaching or lixiviation methods to progressively concentrate a given element, ore, or mineral from a dilute solution containing numerous other ions through a series of chelating and elution stages.
• Extractants used for oxide copper are oximes and modified aldoximes, typically ACORGA® (Solvay) and LIX® (BASF) products. Extractants used for uranium processing are tertiary amines exemplified by the Alamine® reagents made by BASF.
• the third general type of extractants are phosphatederived, such as Di-(2-Ethyl Hexyl) phosphoric acid (D2EHPA), trioctylphosphine oxide (TOPO), and tributyl phosphate (TBP), which are widely commercially available. This third group makes up the largest volume of extractants and are so widely available that they are considered generic.
• phosphine extractants such as Solvay’s CYANEX® and Clariant’ s HostarexTM. This group of chemicals is used for the separation of rare earth oxides, cobalt, platinum group metals, and others. They are used in relatively small quantities and are the most expensive.
• Solvent extractants are used primarily in the hydrometallurgical treatment of copper and uranium ores. Usage in copper mining, in which solvent extraction is particularly attractive for lower-grade ores and workover of tailings material. The hydrometallurgical extraction of copper is particularly attractive today because it is recognized as the most likely technology to yield low operating costs.
• the subject invention relates generally to solvent extraction compositions and methods of using said compositions. More specifically, the subject invention provides environmentally-friendly solvent extraction compositions and methods for solvent extraction, such as, for example, during mining and beneficiation processes. In certain embodiments, existing methods can incorporate the subject compositions and methods.
• compositions and methods of the subject invention increase the efficiency of solvent extraction and can decrease the chemical usage, including chemical surfactant or organic solvent usage, required for solvent extraction. Accordingly, the subject invention can be useful for reducing the time needed for mining and the ensuing beneficiation processes.
• the subject invention provides compositions comprising components that are derived from microorganisms.
• the composition comprises a microbial biosurfactant.
• the composition comprises one or more biosurfactants, and, optionally, other compounds, such as, for example, water; chemical surfactants; organic solvents; oximes; modified aldoximes; amines, including, for example, second and tertiary amines; phosphates; phosphines; chelating agents, including, for example, EDTA; acids; diluents; polymers; or any combination thereof.
• the biosurfactant of the composition is utilized in crude form.
• the crude form can comprise, in addition to the biosurfactant, the fermentation broth in which a biosurfactant-producing microorganism was cultivated, residual microbial cell matter or live or inactive microbial cells, residual nutrients, and/or other microbial growth by-products.
• the biosurfactant is utilized after being extracted from a fermentation broth and, optionally, purified.
• the biosurfactant according to the subject invention can be a glycolipid (e.g., sophorolipid, rhamnolipid, cellobiose lipid, mannosylerythritol lipid or trehalose lipid), lipopeptide (e.g., surfactin, iturin, fengycin, arthrofactin, or lichenysin), flavolipid, phospholipid (e.g., cardiolipins), fatty acid ester compound, fatty acid ether compound, and/or high molecular weight polymers such as lipoproteins, lipopolysaccharide-protein complexes, and polysaccharide-protein-fatty acid complexes.
• a glycolipid e.g., sophorolipid, rhamnolipid, cellobiose lipid, mannosylerythritol lipid or trehalose lipid
• lipopeptide e.g., surfactin, iturin, fengycin
• the subject invention provides a method for solvent extraction, wherein the method comprises the following steps: a) contacting a solvent extraction composition comprising a biosurfactant with a liquid containing an element, metal, mineral, or other substance of interest; and b) recovering the element, metal, mineral, or other substance of interest from the solvent extraction composition.
• the solvent extraction composition according to the subject invention is effective due to improving phase transfer times, reducing crud formation, and/or protecting against extractant losses due to nitration and oxidation.
• the methods of the subject invention result in at least a 25% increase in the recovery of elements, minerals, metals, or other substances of interest, preferably at least a 50% increase, after one treatment.
• the liquid composition can be treated multiple times to further increase the amount of recovered elements, metals, minerals, or other substances of interest.
• applying refers to contacting it with a target or site such that the composition or product can have an effect on that target or site.
• the effect can be due to, for example, microbial growth and/or the action of a biosurfactant or other microbial growth by-product.
• biofilm is a complex aggregate of microorganisms, such as bacteria, yeast, or fungi, wherein the cells adhere to each other and/or to a surface using an extracellular matrix.
• the cells in biofilms are physiologically distinct from planktonic cells of the same organism, which are single cells that can float or swim in liquid medium.
• an “isolated” or “purified” nucleic acid molecule, polynucleotide, polypeptide, protein or organic compound such as a small molecule (e.g., those described below), is substantially free of other compounds, such as cellular material, with which it is associated in nature.
• a purified or isolated polynucleotide ribonucleic acid (RNA) or deoxyribonucleic acid (DNA)
• RNA ribonucleic acid
• DNA deoxyribonucleic acid
• a purified or isolated polypeptide is free of the amino acids or sequences that flank it in its naturally-occurring state.
• An isolated microbial strain means that the strain is removed from the environment in which it exists in nature. Thus, the isolated strain may exist as, for example, a biologically pure culture, or as spores (or other forms of the strain) in association with a carrier.
• purified compounds are at least 60% by weight the compound of interest.
• the preparation is at least 75%, more preferably at least 90%, and most preferably at least 98%, by weight the compound of interest.
• a purified compound is one that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 98%, 99%, or 100% (w/w) of the desired compound by weight. Purity is measured by any appropriate standard method, for example, by column chromatography, thin layer chromatography, or high-performance liquid chromatography (HPLC) analysis.
• HPLC high-performance liquid chromatography
• a “metabolite” refers to any substance produced by metabolism or a substance necerney for taking part in a particular metabolic process.
• a metabolite can be an organic compound that is a starting material, an intermediate in, or an end product of metabolism.
• metabolites include, but are not limited to, enzymes, acids, solvents, alcohols, proteins, vitamins, minerals, microelements, amino acids, biopolymers and biosurfactants.
• a “reduction” means a negative alteration
• an “increase” means a positive alteration, wherein the negative or positive alteration 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%.
• surfactant means a compound that lowers the surface tension (or interfacial tension) between two liquids or between a liquid and a solid. Surfactants act as, e.g., detergents, wetting agents, emulsifiers, foaming agents, and/or dispersants.
• a “biosurfactant” is a surface-active substance produced by a living cell and/or using naturally-derived substrates.
• Biosurfactants are a structurally diverse group of surface-active substances consisting of two parts: a polar (hydrophilic) moiety and non-polar (hydrophobic) group. Due to their amphiphilic structure, biosurfactants can, for example, increase the surface area of hydrophobic water-insoluble substances, increase the water bioavailability of such substances, and change the properties of bacterial cell surfaces. Biosurfactants can also reduce the interfacial tension between water and oil and, therefore, lower the hydrostatic pressure required to move entrapped liquid to overcome the capillary effect. Biosurfactants accumulate at interfaces, thus reducing interfacial tension and leading to the formation of aggregated micellar structures in solution. The formation of micelles provides a physical mechanism to mobilize, for example, oil in a moving aqueous phase.
• biosurfactants to reduce the surface tension also permits their use as antibacterial, antifungal, and hemolytic agents to, for example, control pests and/or microbial growth.
• the hydrophilic group of a biosurfactant is a sugar (e.g., a mono-, di-, or polysaccharide) or a peptide
• the hydrophobic group is typically a fatty acid.
• biosurfactant molecules based on, for example, type of sugar, number of sugars, size of peptides, which amino acids are present in the peptides, fatty acid length, saturation of fatty acids, additional acetylation, additional functional groups, esterification, polarity and charge of the molecule.
• glycolipids e.g., sophorolipids, rhamnolipids, cellobiose lipids, mannosyleiythritol lipids and trehalose lipids
• lipopeptides e.g., surfactin, iturin, fengycin, arthrofactin and lichenysin
• flavolipids e.g., phospholipids (e.g., cardiolipins)
• phospholipids e.g., cardiolipins
• fatty acid ester compounds e.g., and high molecular weight polymers such as lipoproteins, lipopolysaccharide-protein complexes, and polysaccharide-protein-fatty acid complexes.
• Each type of biosurfactant within each class can further comprise subtypes having further modified structures.
• each biosurfactant molecule has its own HLB value depending on its structure; however, unlike production of chemical surfactants, which results in a single molecule with a single HLB value or range, one cycle of biosurfactant production typically results in a mixture of biosurfactant molecules (e.g., subtypes and isomers thereof).
• biosurfactant and “biosurfactant molecule” include all forms, analogs, orthologs, isomers, and natural and/or anthropogenic modifications of any biosurfactant class (e.g., glycolipid) and/or subtype thereof (e.g., sophorolipid).
• biosurfactant class e.g., glycolipid
• subtype thereof e.g., sophorolipid
• SLP sephorolipid
• SLP molecule includes all forms, and isomers thereof, of SLP molecules, including, for example, acidic (linear) SLP (ASL) and lactonic SLP (LSL).
• ASL acidic (linear) SLP
• LSL lactonic SLP
• the glycolipid biosurfactant is a sophorolipid (SLP).
• Sophorolipids are glycolipid biosurfactants produced by, for example, various yeasts of the Starmerella clade when cultivated in the presence of a hydrocarbon-based source of one or more fatty acids.
• SLP typically consist of a disaccharide sophorose linked to long chain hydroxy fatty acids. They can comprise a partially acetylated 2-O-P-D-glucopyranosyl-D-glucopyranose unit attached p-gly cosid ically to 17-L- hydroxyoctadecanoic or 17-L-hydroxy-A9-octadecenoic acid.
• the SLP according to the subject invention are represented by General Formula (1) and/or General Formula (2), and are obtained as a collection of 30 or more types of structural homologs: where R 1 and R 1 ' independently represent saturated hydrocarbon chains or single or multiple, in particular single, unsaturated hydrocarbon chains having 8 to 20, in particular 12 to 18 carbon atoms, more preferably 14 to 18 carbon atoms, which can be linear or branched and can comprise one or more hydroxy groups, R 2 and R 2 independently represent a hydrogen atom or a saturated alkyl functional group or a single or multiple, in particular single, unsaturated alkyl functional group having 1 to 9 carbon atoms, more preferably 1 to 4 carbon atoms, which can be linear or branched and can comprise one or more hydroxy groups, and R 3 , R 3 , R 4 and R 4 independently represent a hydrogen atom or - COCH 3 .
• composition utilized according to the subject methods can comprises more than one form of SLP, including linear SLP and lactonic SLP.
• SLP can be non-acetylated, mono-acetylated and/or di-acetylated SLP.
• the composition comprises SLP according to General Formula (1) (linear SLP) wherein R 1 and/or R 2 are an acetyl group, and wherein R 3 is derived from a stearic, oleic and/or linoleic fatty acid.
• SLP according to General Formula (1) (linear SLP) wherein R 1 and/or R 2 are an acetyl group, and wherein R 3 is derived from a stearic, oleic and/or linoleic fatty acid.
• gangue materials are removed from the substance of interest (e.g., element, compound, mineral).
• the term “about” is understood as within a range of normal tolerance in the art, for example within 2 standard deviations of the mean. About can be understood as within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05%, or 0.01% of the stated value. Unless otherwise clear from context, all numerical values provided herein are modified by the term about.
• the subject invention provides compositions comprising components that are derived from microorganisms.
• the composition comprises a microbial biosurfactant.
• the composition comprises one or more biosurfactants, and, optionally, other compounds, such as, for example, water, chemical surfactants, extractants, acids, organic solvents, oximes, modified aldoximes, amines (e.g., second and tertiary amines), phosphates, phosphines, chelating agents (e.g., EDTA), diluents, polymers, or any combination thereof.
• the polymers can include natural or synthetic polymers, water soluble polymers, cationic polymers, anionic polymers, or non-ionic polymers.
• the polymers can be, for example, anionic polyacrylamide, modified polyacrylamide, nonionic polyacrylamide, starch, guar gum, Moringa oleifera seed extract, Strychnos potatorum seed extract, gelatin (e.g., isinglass), alginate (e.g., sodium alginate), or polymeric ferric sulfate.
• the acid is hydrochloric acid, ascorbic acid, oxalic acid, citric acid, sulfuric acid, or any combination thereof.
• the oxime or modified aldoxime is dimethylglyoxime, salicylaldoxime, 5-nonylsalicylaldoxime, 5-nonyl-2-hydroxyacetophenone oxime, 5- dodecylsalicylaldoxime, an amidoxime (e.g., polyaciylamidoxim), or any combination thereof.
• chelating agents can be, but are not limited to, ethylenediaminetetraacetic acid (EDTA), nitrilotriacetic acid (NTA), nitrilotris(methylene)triphosphonic acid (NTTA), trimethylenedinitrilotetraacetic acid (TMDTA), L-5- glutamyl-L-cysteinylglycine (GCG), calcium trisodium diethylenetriaminepentaacetic acid, sodium nitrilotriacetic acid, a phosphonate, succimer (DMSA), diethylenetriaminepentaacetate (DTP A), N- acetylcysteine, n-hydroxyethylethylenediaminetriacetic acid (HEDTA), organic acids with more than one coordination group (e.g., rubeanic acid), STPP (sodiumtripolyphosphate, NasPsOio), trisodium phosphate (TSP), water, carbohydrates, organic acids with more than one coordination group
• the biosurfactant is utilized after being extracted from a fermentation broth and, optionally, purified.
• the biosurfactant according to the subject invention can be a glycolipid (e.g., sophorolipids, rhamnolipids, cellobiose lipids, mannosylerythritol lipids and trehalose lipids), lipopeptide (e.g., surfactin, iturin, fengycin, arthrofactin and lichenysin), flavolipid, phospholipid (e.g., cardiolipins), fatty acid ester compound, fatty acid ether compound, and/or high molecular weight polymers such as lipoproteins, lipopolysaccharide-protein complexes, and polysaccharide-protein-fatty acid complexes.
• a glycolipid e.g., sophorolipids, rhamnolipids, cellobiose lipids, mannosylerythritol lipids and trehalose lipids
• lipopeptide e.g., surfactin, it
• the biosurfactant is a sophorolipid (SLP), including linear SLP, lactonic SLP, acetylated SLP, de-acetylated SLP, salt-form SLP derivatives, esterified SLP derivatives, amino acid-SLP conjugates, and other SLP derivatives or isomers that exist in nature and/or are produced synthetically.
• SLP sophorolipid
• the SLP is a linear SLP or a derivatized linear SLP.
• the subject compositions can comprise lactonic and linear SLP, with 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 comprising linear forms, and the remainder comprising lactonic forms.
• the biosurfactant can be included in the composition at 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, with respect to the total solvent extraction composition.
• a purified biosurfactant may be added in combination with an acceptable carrier, in that the biosurfactant may be presented at concentrations of 0.001 to 50% (v/v), preferably, 0.01 to 20% (v/v), more preferably, 0.02 to 5% (v/v).
• the biosurfactant can be included in the composition at, for example, 0.01 to 100,000 ppm, 0.05 to 10,000 ppm, 0.1 to 1,000 ppm, 0.5 to 750 ppm, 1.0 to 500 ppm, 2.0 to 250 ppm, or 3.0 to 100 ppm, with respect to the amount of liquid being treated.
• the solvent extraction composition can further comprise other additives such as, for example, carriers, other microbe-based compositions, additional biosurfactants, enzymes, catalysts, solvents, salts, buffers, emulsifying agents, lubricants, solubility controlling agents, preservatives, stabilizers, ultra-violet light resistant agents, viscosity modifiers, preservatives, tracking agents, and other microbes and other ingredients specific for an intended use.
• additives such as, for example, carriers, other microbe-based compositions, additional biosurfactants, enzymes, catalysts, solvents, salts, buffers, emulsifying agents, lubricants, solubility controlling agents, preservatives, stabilizers, ultra-violet light resistant agents, viscosity modifiers, preservatives, tracking agents, and other microbes and other ingredients specific for an intended use.
• the ores are low-grade ores, in which the ores comprise less than about 50%, about 40%, about 35%, about 30%, or about 25% of the substance of interest (e.g., metal, mineral, compound or element being mined), with the remainder comprising gangue.
• substance of interest e.g., metal, mineral, compound or element being mined
• the solvent extraction composition can be applied to a solution containing the element, mineral, metal, or other compound of interest and, optionally, mixed by adding, pouring, shaking or combining.
• the solution and solvent extraction composition are mixed using a mechanical shaker.
• the time period in which the solvent extraction composition can be contacted and/or mixed to a liquid containing the element, mineral, metal, or other compound of interest is for 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 1 hour to about 24 hours.
• the methods comprise applying a liquid or solid form of the solvent extraction composition to the liquid for the period of time in which liquid containing the metal, element, mineral, or other compound of interest is being produced or until the amount of metal, element, mineral, or other compound of interest has been recovered is determined to be satisfactory, which can be readily determined by one skilled in the art.
• the solvent extraction compositions can be used in beneficiation processes, particularly in low-grade ores containing low concentrations of the element, mineral, metal, or other substance of interest, such as, for example, copper, nickel, cobalt, or uranium.
• the element, mineral, metal, or compound of interest it can be necessary to crush and grind the ore and preconcentrate or separate the element or substance of interest from the ore by flotation or gravity separation (i.e., settling).
• the solvent extraction compositions can be used in methods of leaching or as alternatives to methods of leaching, such as, for example, gold cyanidation.
• the process of extraction by leaching includes leaching (e.g., cyanide leaching), washing and filtering of leaching pulp, extraction of the metal from the leaching solution or pulp, and smelting of finished products.
• the solvent extraction compositions can be used instead of cyanide in order to recover gold from gold-bearing ore.
• the processes according to the invention can be applied to any metal, element, mineral, or other compound of interest containing stream. It is advantageously applied to a metal, element, mineral, or other compound of interest containing stream resulting from an existing leaching operation where a metal, element, mineral, or other compound of interest is present in solution.
• the metal, element, mineral, or other compound of interest can be recovered without downstream impact on the leaching operation or other solvent extraction operations. Additionally, a metal, element, mineral, or other compound of interest can be economically recovered without further mining costs since it is already present in the solution.
• different acidic aqueous solutions may be used as the aqueous feed solution, such as leach solutions from existing solvent extraction operations, i.e. copper solvent extraction operations, scrub liquors from acid plants/smelting operations, leach solutions from the processing of flue dusts, filter cakes, metal oxide ores, reprocessing of spent catalysts, or other waste streams containing metals, elements, minerals, or other substances of interests such as, but not limited to, lubricant wastes. More than one source of aqueous solution containing metal can be used.
• the metal containing aqueous feed solution can be acidified (i.e., leached) prior to going to the extraction step in order to limit loading other impurities. While it is common practice in current solvent extraction processes to add an acid to the leach solution (after solvent extraction) to enhance metal dissolution, the subject invention provides adding the acid to the stream of an existing solvent extraction operation prior to, during, and/or after extracting the target metal, mineral, or element in order to enhance selectivity and recovery, and before the leach solution is returned to the primary metal extraction process.
• the solution containing the metal, element, mineral, or other compound of interest can have a pH of less than about 6, less than about 4, less than about 2.5, or less than about 1 .
• the solvent extraction composition according to the subject invention is effective due to improving phase transfer times (i.e., the transfer of the substance of interest from the aqueous phase to the organic phase), reducing crud formation, particularly during settling of the aqueous and organic phases, and/or protecting against extractant losses due to nitration and oxidation.
• phase transfer times i.e., the transfer of the substance of interest from the aqueous phase to the organic phase
• the solvent extraction composition according to the subject invention provides enhanced or increased efficiency of recovering metals, minerals, compounds, or other substances of interest with limited negative environmental impacts. Additionally, the methods of the subject invention do not require complicated equipment or high energy consumption, and the production of the solvent extraction composition can be performed on site, including, for example, at a mine or at an industrial site. In certain embodiments, the subject solvent extraction composition can result in a decreased use of chemical surfactants, synthetic solvent extraction agents, or other potentially harmful chemicals used for solvent extraction.
• the subject invention provides methods for cultivation of microorganisms and production of microbial metabolites and/or other by-products of microbial growth.
• the subject invention further utilizes cultivation processes that are suitable for cultivation of microorganisms and production of microbial metabolites on a desired scale. These cultivation processes include, but are not limited to, submerged cultivation/fermentation, solid state fermentation (SSF), and modifications, hybrids and/or combinations thereof.
• SSF solid state fermentation
• the microorganisms can be, for example, bacteria, yeast and/or fungi. These microorganisms may be natural, or genetically modified microorganisms. For example, the microorganisms may be transformed with specific genes to exhibit specific characteristics.
• the microorganisms may also be mutants of a desired strain.
• “mutant” means a strain, genetic variant or subtype of a reference microorganism, wherein the mutant has one or more genetic variations (e.g., a point mutation, missense mutation, nonsense mutation, deletion, duplication, frameshift mutation or repeat expansion) as compared to the reference microorganism. Procedures for making mutants are well known in the microbiological art. For example, UV mutagenesis and nitrosoguanidine are used extensively toward this end.
• the microbes are capable of producing amphiphilic molecules, enzymes, proteins and/or biopolymers.
• Microbial biosurfactants are produced by a variety of microorganisms such as bacteria, fungi, and yeasts, including, for example, Agrobacterium spp. (e.g., A. radiobacter),- Arthrobacter spp.; Aspergillus spp.; Aureobasidium spp. (e.g., A. pullulans .
• Azotobacter e.g., A. vinelandii, A. chroococcuniy, Azospirillum spp. (e.g., A. brasiliensis),- Bacillus spp.
• Candida spp. e.g., C. albicans, C. rugosa, C. tropicalis, C. lipolytica, C. lorulopsisy.
• Clostridium e.g., C. butyricum, C. tyrobutyricum, C. acetobutyricum, and C.
• Pichia spp. e.g., P. anomala, P. guilliermondii, P. occidentalis, P. kudriavzeviiy. Phycomyces spp.; Phythium spp.
• Ralslonia spp. e.g., R. eulrophd , Rhodococcus spp. (e.g., R. erythropolisy, Rhodospirillum spp. (e.g., R. rubrum); Rhizobium spp.; Rhizopus spp.; Saccharomyces spp. (e.g., >S’. cerevisiae, S. boulardii sequela, S. torula Sphingomonas spp. (e.g., S. paucimobilis); Starmerella spp. (e.g., 5.
• growth refers to cultivation or growth of cells under controlled conditions.
• the growth could be aerobic or anaerobic.
• the microorganisms are grown using SSF and/or modified versions thereof.
• the vessel may also be able to monitor the growth of microorganisms inside the vessel (e.g., measurement of cell number and growth phases).
• a daily sample may be taken from the vessel and subjected to enumeration by techniques known in the art, such as dilution plating technique.
• Dilution plating is a simple technique used to estimate the number of organisms in a sample. The technique can also provide an index by which different environments or treatments can be compared.
• the method includes supplementing the cultivation with a nitrogen source.
• the nitrogen source can be, for example, potassium nitrate, ammonium nitrate ammonium sulfate, ammonium phosphate, ammonia, urea, and/or ammonium chloride. These nitrogen sources may be used independently or in a combination of two or more.
• the method can provide oxygenation to the growing culture.
• One embodiment utilizes slow motion of air to remove low-oxygen containing air and introduce oxygenated air.
• the oxygenated air may be ambient air supplemented daily through mechanisms including impellers for mechanical agitation of liquid, and air spargers for supplying bubbles of gas to liquid for dissolution of oxygen into the liquid.
• Usable inorganic salts can 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 independently or in a combination of two or more.
• the method for cultivation of microorganisms is carried out at about 5° to about 100° C, preferably, 15 to 60° C, more preferably, 25 to 50° C.
• the cultivation may be carried out continuously at a constant temperature.
• the cultivation may be subject to changing temperatures.
• the biomass content of the fermentation medium may be, for example, from 5 g/1 to 180 g/1 or more, or from 10 g/1 to 150 g/1.
• the cell concentration may be, for example, at least 1 x 10 6 to 1 x 10 12 , 1 x 10 7 to 1 x 10 11 , 1 x 10 8 to 1 x 10 10 , or 1 x 10 9 CFU/ml.
• all of the microbial cultivation composition is removed upon the completion of the cultivation (e.g., upon, for example, achieving a desired cell density, or density of a specified metabolite).
• this batch procedure an entirely new batch is initiated upon harvesting of the first batch.
• biomass with viable cells, spores, conidia, hyphae and/or mycelia remains in the vessel as an inoculant for a new cultivation batch.
• the composition that is removed can be a cell-free medium or contain cells, spores, or other reproductive propagules, and/or a combination of thereof. In this manner, a quasi-continuous system is created.
• the method does not require complicated equipment or high energy consumption.
• the microorganisms of interest can be cultivated at small or large scale on site and utilized, even being still-mixed with their media.
• the subject invention provides a “microbe-based composition,” meaning a composition that comprises components that were produced as the result of the growth of microorganisms or other cell cultures.
• the microbe-based composition may comprise the microbes themselves and/or by-products of microbial growth.
• the microbes may be in a vegetative state, in spore form, in mycelial form, in any other form of propagule, or a mixture of these.
• the microbes may be planktonic or in a biofilm form, or a mixture of both.
• the by-products of growth may be, for example, metabolites, cell membrane components, expressed proteins, and/or other cellular components.
• the microbes may be intact or lysed.
• the subject invention further provides “microbe-based products,” which are products that are to be applied in practice to achieve a desired result.
• the microbe-based product can be simply a microbe-based composition harvested from the microbe cultivation process.
• the microbe-based product may comprise further ingredients that have been added. These additional ingredients can include, for example, stabilizers, acids, buffers, carriers, such as water, salt solutions, or any other appropriate carrier, added nutrients to support further microbial growth, non-nutrient growth enhancers, and/or agents that facilitate tracking of the microbes and/or the composition in the environment to which it is applied.
• the microbe-based product may also comprise mixtures of microbebased compositions.
• the microbe-based product may also comprise one or more components of a microbe-based composition that have been processed in some way such as, but not limited to, filtering, centrifugation, lysing, diying, purification and the like.
• One microbe-based product of the subject invention is simply the fermentation medium containing the microorganisms and/or the microbial metabolites produced by the microorganisms and/or any residual nutrients.
• the product of fermentation may be used directly without extraction or purification. If desired, extraction and purification can be easily achieved using standard extraction and/or purification methods or techniques described in the literature.
• microorganisms in the microbe-based products may be in an active or inactive form, or in the form of vegetative cells, reproductive spores, conidia, mycelia, hyphae, or any other form of microbial propagule.
• the microbe-based products may also contain a combination of any of these forms of a microorganism.

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• Manufacturing & Machinery (AREA)
• General Life Sciences & Earth Sciences (AREA)
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• Organic Chemistry (AREA)
• Chemical Kinetics & Catalysis (AREA)
• General Chemical & Material Sciences (AREA)
• Biochemistry (AREA)
• Biotechnology (AREA)
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• Extraction Or Liquid Replacement (AREA)
• Micro-Organisms Or Cultivation Processes Thereof (AREA)
• Manufacture And Refinement Of Metals (AREA)

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• 2023
  • 2023-07-27 CN CN202380077874.4A patent/CN120167014A/zh active Pending
  • 2023-07-27 AU AU2023341918A patent/AU2023341918A1/en active Pending
  • 2023-07-27 EP EP23866305.8A patent/EP4587600A1/de active Pending
  • 2023-07-27 CA CA3267493A patent/CA3267493A1/en active Pending
  • 2023-07-27 WO PCT/US2023/071119 patent/WO2024059371A1/en not_active Ceased
  • 2023-07-27 PE PE2025000550A patent/PE20251182A1/es unknown
• 2025
  • 2025-03-12 CL CL2025000692A patent/CL2025000692A1/es unknown
  • 2025-03-12 MX MX2025002928A patent/MX2025002928A/es unknown

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