WO2024030815A1 - Emulsion explosive compositions and methods of use - Google Patents
Emulsion explosive compositions and methods of use Download PDFInfo
- Publication number
- WO2024030815A1 WO2024030815A1 PCT/US2023/071109 US2023071109W WO2024030815A1 WO 2024030815 A1 WO2024030815 A1 WO 2024030815A1 US 2023071109 W US2023071109 W US 2023071109W WO 2024030815 A1 WO2024030815 A1 WO 2024030815A1
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- WIPO (PCT)
- Prior art keywords
- mine
- composition
- emulsion explosive
- biosurfactant
- sophorolipid
- Prior art date
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- MNNHAPBLZZVQHP-UHFFFAOYSA-N diammonium hydrogen phosphate Chemical compound [NH4+].[NH4+].OP([O-])([O-])=O MNNHAPBLZZVQHP-UHFFFAOYSA-N 0.000 description 1
- LFMYQKSTJULFQX-UHFFFAOYSA-N diazanium nitric acid sulfate Chemical compound [NH4+].[NH4+].O[N+]([O-])=O.[O-]S([O-])(=O)=O LFMYQKSTJULFQX-UHFFFAOYSA-N 0.000 description 1
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 1
- 238000007598 dipping method Methods 0.000 description 1
- 150000002016 disaccharides Chemical class 0.000 description 1
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 1
- 239000006185 dispersion Substances 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- OAEGRYMCJYIXQT-UHFFFAOYSA-N dithiooxamide Chemical compound NC(=S)C(N)=S OAEGRYMCJYIXQT-UHFFFAOYSA-N 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 229920001971 elastomer Polymers 0.000 description 1
- 239000003623 enhancer Substances 0.000 description 1
- 239000003797 essential amino acid Substances 0.000 description 1
- 235000020776 essential amino acid Nutrition 0.000 description 1
- 210000002744 extracellular matrix Anatomy 0.000 description 1
- 239000004744 fabric Substances 0.000 description 1
- 239000003925 fat Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 231100000221 frame shift mutation induction Toxicity 0.000 description 1
- 230000037433 frameshift Effects 0.000 description 1
- 239000001530 fumaric acid Substances 0.000 description 1
- 235000011087 fumaric acid Nutrition 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 230000007614 genetic variation Effects 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 229960003180 glutathione Drugs 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 125000003147 glycosyl group Chemical group 0.000 description 1
- 239000003102 growth factor Substances 0.000 description 1
- 239000003219 hemolytic agent Substances 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 229930195733 hydrocarbon Natural products 0.000 description 1
- 150000002430 hydrocarbons Chemical class 0.000 description 1
- 125000001165 hydrophobic group Chemical group 0.000 description 1
- 230000002706 hydrostatic effect Effects 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 239000002555 ionophore Substances 0.000 description 1
- 230000000236 ionophoric effect Effects 0.000 description 1
- FBAFATDZDUQKNH-UHFFFAOYSA-M iron chloride Chemical compound [Cl-].[Fe] FBAFATDZDUQKNH-UHFFFAOYSA-M 0.000 description 1
- 229910000358 iron sulfate Inorganic materials 0.000 description 1
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 description 1
- 239000010443 kyanite Substances 0.000 description 1
- 229910052850 kyanite Inorganic materials 0.000 description 1
- 239000004310 lactic acid Substances 0.000 description 1
- 235000014655 lactic acid Nutrition 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- HWSZZLVAJGOAAY-UHFFFAOYSA-L lead(II) chloride Chemical compound Cl[Pb]Cl HWSZZLVAJGOAAY-UHFFFAOYSA-L 0.000 description 1
- 239000000944 linseed oil Substances 0.000 description 1
- 235000021388 linseed oil Nutrition 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 230000002934 lysing effect Effects 0.000 description 1
- 229910001629 magnesium chloride Inorganic materials 0.000 description 1
- 235000011147 magnesium chloride Nutrition 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 239000001630 malic acid Substances 0.000 description 1
- 235000011090 malic acid Nutrition 0.000 description 1
- 239000011565 manganese chloride Substances 0.000 description 1
- 235000002867 manganese chloride Nutrition 0.000 description 1
- 229940099607 manganese chloride Drugs 0.000 description 1
- 229940099596 manganese sulfate Drugs 0.000 description 1
- 239000011702 manganese sulphate Substances 0.000 description 1
- 235000007079 manganese sulphate Nutrition 0.000 description 1
- WPBNNNQJVZRUHP-UHFFFAOYSA-L manganese(2+);methyl n-[[2-(methoxycarbonylcarbamothioylamino)phenyl]carbamothioyl]carbamate;n-[2-(sulfidocarbothioylamino)ethyl]carbamodithioate Chemical compound [Mn+2].[S-]C(=S)NCCNC([S-])=S.COC(=O)NC(=S)NC1=CC=CC=C1NC(=S)NC(=O)OC WPBNNNQJVZRUHP-UHFFFAOYSA-L 0.000 description 1
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 235000012054 meals Nutrition 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 229910021645 metal ion Inorganic materials 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 235000010755 mineral Nutrition 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- 231100000350 mutagenesis Toxicity 0.000 description 1
- 238000002703 mutagenesis Methods 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- MGFYIUFZLHCRTH-UHFFFAOYSA-N nitrilotriacetic acid Chemical compound OC(=O)CN(CC(O)=O)CC(O)=O MGFYIUFZLHCRTH-UHFFFAOYSA-N 0.000 description 1
- 235000021095 non-nutrients Nutrition 0.000 description 1
- 230000037434 nonsense mutation Effects 0.000 description 1
- 125000001400 nonyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- ZQPPMHVWECSIRJ-KTKRTIGZSA-N oleic acid group Chemical group C(CCCCCCC\C=C/CCCCCCCC)(=O)O ZQPPMHVWECSIRJ-KTKRTIGZSA-N 0.000 description 1
- 239000004006 olive oil Substances 0.000 description 1
- 235000008390 olive oil Nutrition 0.000 description 1
- 238000006213 oxygenation reaction Methods 0.000 description 1
- 239000003002 pH adjusting agent Substances 0.000 description 1
- 239000003973 paint Substances 0.000 description 1
- 239000000123 paper Substances 0.000 description 1
- UEZVMMHDMIWARA-UHFFFAOYSA-M phosphonate Chemical compound [O-]P(=O)=O UEZVMMHDMIWARA-UHFFFAOYSA-M 0.000 description 1
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 1
- 239000011505 plaster Substances 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 235000020777 polyunsaturated fatty acids Nutrition 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 239000004323 potassium nitrate Substances 0.000 description 1
- 235000010333 potassium nitrate Nutrition 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- BDERNNFJNOPAEC-UHFFFAOYSA-N propan-1-ol Chemical compound CCCO BDERNNFJNOPAEC-UHFFFAOYSA-N 0.000 description 1
- 235000019260 propionic acid Nutrition 0.000 description 1
- 229940107700 pyruvic acid Drugs 0.000 description 1
- IUVKMZGDUIUOCP-BTNSXGMBSA-N quinbolone Chemical compound O([C@H]1CC[C@H]2[C@H]3[C@@H]([C@]4(C=CC(=O)C=C4CC3)C)CC[C@@]21C)C1=CCCC1 IUVKMZGDUIUOCP-BTNSXGMBSA-N 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 239000008165 rice bran oil Substances 0.000 description 1
- 229910052895 riebeckite Inorganic materials 0.000 description 1
- 229920002477 rna polymer Polymers 0.000 description 1
- 239000011435 rock Substances 0.000 description 1
- 239000005060 rubber Substances 0.000 description 1
- 229930195734 saturated hydrocarbon Natural products 0.000 description 1
- 239000008159 sesame oil Substances 0.000 description 1
- 235000011803 sesame oil Nutrition 0.000 description 1
- 238000010008 shearing Methods 0.000 description 1
- 229910000029 sodium carbonate Inorganic materials 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- 239000000176 sodium gluconate Substances 0.000 description 1
- 235000012207 sodium gluconate Nutrition 0.000 description 1
- 229940005574 sodium gluconate Drugs 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- PZDOWFGHCNHPQD-VNNZMYODSA-N sophorose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](C=O)O[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O PZDOWFGHCNHPQD-VNNZMYODSA-N 0.000 description 1
- 125000003152 sophorose group Chemical group 0.000 description 1
- 235000020712 soy bean extract Nutrition 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 239000010959 steel Substances 0.000 description 1
- 150000003431 steroids Chemical class 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 229960005346 succimer Drugs 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 238000004809 thin layer chromatography Methods 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- VZCYOOQTPOCHFL-UHFFFAOYSA-N trans-butenedioic acid Natural products OC(=O)C=CC(O)=O VZCYOOQTPOCHFL-UHFFFAOYSA-N 0.000 description 1
- 238000011282 treatment Methods 0.000 description 1
- 125000004417 unsaturated alkyl group Chemical group 0.000 description 1
- 229930195735 unsaturated hydrocarbon Natural products 0.000 description 1
- 230000035899 viability Effects 0.000 description 1
- 239000002023 wood Substances 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 description 1
- 229910000368 zinc sulfate Inorganic materials 0.000 description 1
- 229960001763 zinc sulfate Drugs 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C06—EXPLOSIVES; MATCHES
- C06B—EXPLOSIVES OR THERMIC COMPOSITIONS; MANUFACTURE THEREOF; USE OF SINGLE SUBSTANCES AS EXPLOSIVES
- C06B47/00—Compositions in which the components are separately stored until the moment of burning or explosion, e.g. "Sprengel"-type explosives; Suspensions of solid component in a normally non-explosive liquid phase, including a thickened aqueous phase
- C06B47/14—Compositions in which the components are separately stored until the moment of burning or explosion, e.g. "Sprengel"-type explosives; Suspensions of solid component in a normally non-explosive liquid phase, including a thickened aqueous phase comprising a solid component and an aqueous phase
- C06B47/145—Water in oil emulsion type explosives in which a carbonaceous fuel forms the continuous phase
-
- C—CHEMISTRY; METALLURGY
- C06—EXPLOSIVES; MATCHES
- C06B—EXPLOSIVES OR THERMIC COMPOSITIONS; MANUFACTURE THEREOF; USE OF SINGLE SUBSTANCES AS EXPLOSIVES
- C06B23/00—Compositions characterised by non-explosive or non-thermic constituents
Definitions
- Conventional emulsion explosives are water-in-oil emulsions, comprising inorganic oxidizer salt solution droplets (i.e., discontinuous phase) dispersed in a fuel (i.e., continuous phase).
- the droplets which constitute a dispersion or emulsion phase, are held in place by a water-in-oil emulsifier provided the emulsified state remains stable.
- an emulsifier or surfactant is used in the emulsion for promoting formation and stability of the oxidizer salt solution droplets.
- the detonation velocity characteristic of emulsion explosives makes this class of explosives particularly useful.
- the useful explosive characteristics are based on the mixing of the emulsion explosive.
- the continuous phase of fuel surrounds each discontinuous phase droplet of inorganic oxidizer, providing a large interfacial surface area between the two phases.
- Emulsion explosives have relatively low densities, which limits the detonation pressure of the explosive.
- the oxidizers used in emulsion explosives can crystallize at ambient air temperatures and must often be kept warmed. If the oxidizer crystallizes, the crystal structure can puncture the oxidizer-containing droplets, causing the droplets to collapse and the crystals to agglomerate. Eventually, the emulsion is destabilized and the usefulness of the emulsion explosive is reduced, possibly to the point in which the emulsion explosive is not detonatable.
- the subject invention relates generally to the emulsion explosives. More specifically, the subject invention provides environmentally-friendly compositions and explosion methods, such as, for example, exploding geological formations at mining sites, construction sites, or infrastructure sites. In certain embodiments, the explosion emulsion compositions can be stabilized with the subject compositions and methods.
- the compositions and methods of the subject invention increase the effectiveness of emulsion explosives and can decrease the chemical usage, including chemical surfactant, used for emulsion explosive compositions. Accordingly, the subject invention can be useful for reducing the pollution produced during explosions, including during mining operations, creation of roadways, and construction.
- 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; solid fuels, including, for example, aluminum particles, carbonaceous materials (e.g., gilsonite or coal), vegetable grain (e.g., wheat), and sulfur; liquid fuels, including, for example, mineral oil, silicone oil, waxes, paraffin oils, esters (e.g., dioctyl adipate and isodecyl pelargonate), petroleum distillates (e.g., gasoline, kerosene, and diesel), vegetable oils, (e.g., corn oil, cottonseed oil, peanut oil, and soybean oil), aliphatic or aromatic nitro-compounds, and chlorinated hydrocarbons; cross-linkers; oxidizers; inorganic
- the biosurfactant of the composition is utilized in crude form.
- the crude form can comprise, in addition to the biosurfactant, fermentation broth in which a biosurfactantproducing 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., 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,
- the biosurfactant is a sophorolipid (SLP), including linear SLP, lactonic SLP, acetylated SLP, de-acetylated SLP, salt-form SLP, esterified SLP derivatives, amino acid-SLP conjugates, and other SLP derivatives or isomers that exist in nature and/or are produced synthetically.
- SLP is a linear SLP or a derivatized linear SLP.
- the subject invention provides a method of exploding an object, wherein the method comprises the following steps: a) contacting an emulsion explosive composition according to the subject invention to an object and/or surface; and b) initiating an explosion of the emulsion explosive composition.
- the method reduces the crystallization of oxidizers in the emulsion explosive composition.
- methods of the subject invention comprise contacting an emulsion explosive composition comprising a biosurfactant and, optionally, other components, such as, for example, water; chemical surfactants; solid fuels, including, for example, aluminum particles, carbonaceous materials (e.g., gilsonite or coal), vegetable grain (e.g., wheat), and sulfur; liquid fuels, including, for example, mineral oil, silicone oil, waxes, paraffin oils, esters (e.g., dioctyl adipate and isodecyl pelargonate), petroleum distillates (e.g., gasoline, kerosene, and diesel), vegetable oils, (e.g., com oil, cottonseed oil, peanut oil, and soybean oil), aliphatic or aromatic nitro-compounds, and chlorinated hydrocarbons; cross-linkers; oxidizers; inorganic salts, including, for example, ammonium, potassium, and sodium salts of perchlorates and nitrate;
- the chemical surfactant of the emulsion explosive composition is a detergent, wetting agent, emulsifier, foaming agent, and/or dispersant.
- the method enhances the homogeneity of suspended droplets in the emulsion explosive composition and can increase the surface area of the said droplets.
- the resulting droplets can be less than about 10 cm, about 1 cm, about 1 mm, about 500 pm, about 100 pm, about 10 pm, about 1 pm, about 100 nm, about 10 nm, or about 1 nm in diameter.
- the effectiveness of emulsion explosive results from the surfactant contacting and stabilizing the discontinuous droplets in the emulsion explosive compositions.
- This surfactant allows the oxidizer droplets to remain as a liquid, particularly at ambient air temperature.
- the methods of the subject invention result in at least a 25% increase in the stability of the oxidizer droplets in the composition, preferably at least a 50% increase.
- the emulsion explosive composition according to the subject invention can be effective at inhibiting the crystallization of oxidizers.
- the methods of the subject invention do not require complicated equipment or high energy consumption, and production of the composition can be performed on site, for example, at a mine or at a roadway.
- the subject invention relates generally to emulsion explosive compositions and methods of use. More specifically, the subject invention provides environmentally-friendly compositions and methods for emulsion explosives, such as, for example, for use at mining sites, construction sites, and roadways. In certain embodiments, the emulsion explosive composition can be stabilized with the subject compositions and methods.
- the subject invention is useful for improving the efficiency and efficacy of emulsion explosive compositions.
- the compositions and methods of the subject use safe, environmentally-friendly compositions.
- 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 necessary 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.
- Ranges provided herein are understood to be shorthand for all of the values within the range.
- a range of 1 to 20 is understood to include any number, combination of numbers, or subrange from the group consisting 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 intervening decimal values between the aforementioned integers such as, for example, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, and 1.9.
- “nested sub-ranges” that extend from either end point of the range are specifically contemplated.
- a nested sub-range of an exemplary range of 1 to 50 may comprise 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 the other direction.
- 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, mannosylerythritol 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).
- biosurfactanf 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
- mono-acetylated SLP di-acetylated SLP
- esterified SLP SLP with varying hydrophobic chain lengths
- cationic and/or anionic SLP with fatty acid-amino acid complexes attached esterified SLP
- SLP-metal complexes SLP-salt derivatives (e.g., a sodium salt of a linear SLP), and other, including those that are and/or are not described within in this disclosure.
- the glycolipid biosurfactant is a sophorolipid (SLP).
- 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 0-glycosidically to 17-L- hydroxyoctadecanoic or 17-L-hydroxy-A9-octadecenoic acid.
- the hydroxy fatty acid is generally 16 or 18 carbon atoms, and may contain one or more unsaturated bonds. Furthermore, the sophorose residue can be acetylated on the 6- and/or 6’-position(s).
- the fatty acid carboxyl group can be free (acidic or linear form (General Formula 2)) or internally esterified at the 4"-position (lactonic form (General Formula 1 )).
- S. bombicola produces a specific enzyme, called S. bombicola lactone esterase, which catalyzes the esterification of linear SLP to produce lactonic SLP.
- 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 are typically produced by yeasts, such as Starmerella spp. yeasts and/or Candida spp. yeasts, e.g., Starmerella (Candida) bombicola, Candida apicola, Candida batistae, Candida floricola, Candida riodocensis, Candida stellate and/or Candida kuoi.
- SLP have environmental compatibility, high biodegradability, low toxicity, high selectivity and specific activity in a broad range of temperature, pH and salinity conditions. Additionally, in some embodiments, SLP can be advantageous due to their small micelle size, which can help facilitate the movement of the micelle, and compounds enclosed therein, through nanoscale pores and spaces.
- the micelle size of a 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.
- the glycolipid is a rhamnolipid.
- Rhamnolipids comprise a glycosyl head group (i.e., a rhamnose) moiety, and a 3-(hydroxyalkanoyloxy)alkanoic acid (HAA) fatty acid tail, such as, e.g., 3-hydroxydecanoic acid.
- HAA 3-(hydroxyalkanoyloxy)alkanoic acid
- the HAA moiety can vary in length and degree of branching, depending on, for example, the growth medium and the environmental conditions.
- the highest accumulation of rhamnolipids (RLP) has been shown by submerged cultivation of Pseudomonas spp., such as P. aeruginosa.
- Rhamnolipids according to the subject invention can have the following structure, according to General Formula (3): wherein m is 2, 1 or 0, n is 1 or 0,
- R 1 and R 2 are, independently of one another, the same or a different organic functional group having 2 to 24, preferably 5 to 13 carbon atoms, in particular a substituted or unsubstituted, branched or unbranched alkyl functional group, which can also be unsaturated, wherein the alkyl functional group is a linear saturated alkyl functional group having 8 to 12 carbon atoms, or is a nonyl or a decyl functional group or a mixture thereof. Salts of these compounds are also included according to the invention.
- the term “di-rhamnolipid” is understood to mean compounds of the above formula or the salts thereof in which n is 1.
- “mono-rhamnolipid” is understood in the present invention to mean compounds of the general formula or the salts thereof in which n is 0.
- the composition comprises a mixture of mono- and di-rhamnolipids.
- emulsion explosive refers to a composition comprising two immiscible liquids, such as, for example, oil and water, which is formed by shearing individual droplets (i.e., a discontinuous phase) into a continuous phase.
- the “continuous phase,” as used herein, is usually a liquid fuel (e.g., oil), while the “discontinuous phase,” as used herein is usually an oxidizer.
- Emulsion explosive can further comprise a surfactant to stabilize the droplets in the continuous phase.
- detonation velocity is the velocity at which a shock wave travels through a detonated explosive.
- 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.
- 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 a claim to the specified materials or steps “and those that do not materially affect the basic and novel characteristic(s)” of the claimed invention.
- Use of the term “comprising” contemplates other embodiments that “consist” or “consist essentially of’ the recited component(s).
- 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 emulsion explosive compositions comprising components that are derived from microorganisms.
- the emulsion explosive composition comprises a microbial biosurfactant.
- the composition comprises a biosurfactant, and, optionally, water, chemical surfactants, solid fuels, including, for example, aluminum particles, carbonaceous materials (e.g., gilsonite or coal), vegetable grain (e.g., wheat), and sulfur; liquid fuels, including, for example, mineral oil, silicone oil, waxes, paraffin oils, esters (e.g., dioctyl adipate and isodecyl pelargonate), petroleum distillates (e.g., gasoline, kerosene, and diesel), vegetable oils, (e.g., com oil, cottonseed oil, peanut oil, and soybean oil), aliphatic or aromatic nitro-com pounds, and chlorinated hydrocarbons; cross-linkers
- the emulsion explosive composition comprises a microbe-based product comprising a biosurfactant utilized in crude form.
- the crude form can comprise, in addition to the biosurfactant, 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 product may be, for example, at least, by weight, 1%, 5%, 10%, 25%, 50%, 75%, or 100% broth.
- the amount of biomass in the product, by weight may be, for example, anywhere from 0% to 100% inclusive of all percentages therebetween.
- 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 emulsion explosive 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 the object and/or surface being treated.
- the chemical surfactant of the emulsion explosive composition is a detergent, wetting agent, emulsifier, foaming agent, and/or dispersant.
- the chemical surfactant 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 emulsion explosive composition.
- the concentrations of the fuel of the continuous phase and liquid oxidizer of the discontinuous phase may be vary depending on the exact components of the phases and any other additives.
- the fuel is about 4 wt% to about 30 wt% or about 5 wt% to about 15 wt% of the emulsion explosive.
- the liquid oxidizer comprises about 70 wt% to about 98 wt% or about 85 wt% to about 95 wt% of the composition.
- the biosurfactant comprises, preferably, about 0.25 wt% to about 5 wt% of the composition.
- the solid fuel is aluminum particles, carbonaceous materials (e.g., gilsonite or coal), vegetable grain (e.g., wheat), or sulfur.
- carbonaceous materials e.g., gilsonite or coal
- vegetable grain e.g., wheat
- sulfur e.g., sulfur
- the liquid fuels are mineral oil, silicone oil, waxes, paraffin oils, esters (e.g., dioctyl adipate and isodecyl pelargonate), petroleum distillates (e.g., gasoline, kerosene, and diesel), vegetable oils, (e.g., com oil, cottonseed oil, peanut oil, and soybean oil), aliphatic or aromatic nitro-compounds, or chlorinated hydrocarbons.
- the inorganic salts are ammonium, potassium, or sodium salts of perchlorates or nitrate.
- the emulsifiers are sorbitan monooleate, tartaric acid, isopropyl esters of lanolin fatty acids, substituted oxazalines, or polyisobutyl succinic anhydride (PIBSA).
- PIBSA polyisobutyl succinic anhydride
- the emulsion explosive composition can further comprise other additives such as, for example, carriers, other microbe-based compositions, additional biosurfactants, enzymes, catalysts, solvents, organic salts, buffers, chelating agents, acids, lubricants, solubility controlling agents, preservatives, stabilizers, ultra-violet light resistant agents, viscosity modifiers, preservatives, tracking agents, biocides, 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, organic salts, buffers, chelating agents, acids, lubricants, solubility controlling agents, preservatives, stabilizers, ultra-violet light resistant agents, viscosity modifiers, preservatives, tracking agents, biocides, and other microbes and other ingredients specific for an intended use.
- chelating agents can be, but are not limited to, ethylenediaminetetraacetic acid (EDTA), nitrilotriacetic acid (NTA), a phosphonate, succimer (DMSA), diethylenetriaminepentaacetate (DTPA), A'-acetylcysteine, n- hydroxyethylethylenediaminetriacetic acid (HEDTA), organic acids with more than one coordination group (e.g., rubeanic acid), STPP (sodiumtripolyphosphate, Na5P3O10), trisodium phosphate (TSP), water, carbohydrates, organic acids with more than one coordination group (e.g., citric acid), lipids, steroids, amino acids or related compounds (e.g., glutathione), peptides, phosphates, nucleotides, tetrapyrrols, ferrioxamines, ionophores, orphenolics, sodium citrate, sodium gluconate,
- EDTA
- the emulsion explosives of embodiments of the present invention may be formulated using conventional procedures known in the art.
- the liquid oxidizer, fuel, and biosurfactant solution are approximately equal in temperature when combined.
- the temperature is preferably in a range at which the oxidizer will remain in a liquid state. Accordingly, the selected temperature will be dependent upon the particular oxidizer selected.
- the resulting mixture can be stirred or otherwise mixed vigorously to produce an emulsion of the liquid oxidizer in a continuous liquid fuel phase. Stirring should be continued until the formulation is uniform.
- the formulation process also can be accomplished in a continuous manner as is known in the art. Various modifications to the above-described technique are possible.
- the biosurfactant can be dissolved in the liquid organic fuel prior to combining the organic fuel with the liquid oxidizer to form an emulsion.
- This method allows the emulsion to form quickly and without excessive agitation.
- the emulsifier may be added separately as a third component if desired, may be combined with the liquid oxidizer, or may be combined with the liquid oxidizer and the liquid organic fuel independently before the mixing of the liquid oxidizer and the liquid organic fuel.
- the subject invention provides a method for initiating explosions at various sites, including mining sites, tunneling sites, quarrying sites, construction sites, and roadways.
- the emulsion explosives can be used to displace geological formations and existing human-derived constructions, such as, for example, infrastructure, roadways, buildings, and bridges. The described elements of the subject invention are not an exhaustive examination of all applications.
- the mining site can be a coal mine, iron ore mine (e.g., taconite), copper mine, copper-nickel mine, tin mine, nickel mine, gold mine, silver mine, molybdenum mine, aluminum mine (e.g., bauxite mine, kyanite mine), lead-zine mine, tungsten mine, phosphate mine, potash mine, mica mine, bentonite mine, or zinc mine.
- the mine can be an underground mine, surface mine, placer mine or in situ mine.
- the roadway can be asphalt, tar and chip, earthen roads (e.g., dirt and gravel), or concrete.
- the road surfaces can be intended for use for semi-trucks or other industrial machinery, including for example, excavator or dump trucks, or for use as an automobile, bicycle, or motorcycle racetrack.
- the construction site can be the site of producing a commercial building, including, for example, an office or warehouse; residential building, including, for example, a single family or multifamily home; an industrial facility, including for example, a factory; or an infrastructure project, such as, for example, a roadway, bridge, waterway, or sewer.
- the quarrying site can extract chalk, clay, cinder, coal, sand, gravel, coquina, diabase, gabbro, granite, gritstone, gypsum, limestone, marble, ores, phosphate rock, quartz, sandstone, slate, travertine, or any combination thereof.
- the microbe-containing and/or biosurfactant-containing composition can improve the stability of the oxidizer droplets (i.e., inhibit or reduce crystallization of the oxidizer) within the continuous phase (e.g., fuel), particularly at a range of temperatures including less about 150°C, about 125°C, about 100°C, about 90°C, about 80°C, about 75°C, about 70°C, about 65°C, about 60°C, about 55°C, about 50°C, about 45°C, about 40°C, about 35°C, about 32°C, about 30°C, or lower.
- the continuous phase e.g., fuel
- the microbe-containing and/or biosurfactant-containing composition can stabilize droplets suspended (i.e., discontinuous phase) in a liquid continuous phase. This also allows for an increase in the total surface area of all the droplets within the continuous phase. This increase of the surface area of the droplets allows the for a more efficient explosion, yielding a greater detonation velocity.
- the detonation velocity can be about 18,000 to about 25,000 feet per second (5490 m/s to 7620 m/s), which is determined by the amount of surface area of the discontinuous phase.
- the continuous phase of fuel surrounds each discontinuous phase droplet of inorganic oxidizer, providing a large interfacial surface area between the two phases.
- the composition can reduce the effects of cool temperature or the presence of water on the effectiveness of the emulsion explosives, particularly during blasting.
- the microbe-containing and/or biosurfactant-containing composition has increased longevity versus traditional emulsion explosive compositions due to its reduced formation of crystals in the oxidizer droplets.
- the emulsion explosive composition according to the subject invention is effective due to amphiphiles-mediated droplet stability in a continuous phase.
- the sophorolipid or other biosurfactant serves as a means for facilitating the strength of interfacial film in the composition. The interfacial film forms between the two immiscible phases of the composition: the continuous phase and the discontinuous phase.
- a sophorolipid will form a micelle containing the oxidizer droplet (i.e., discontinuous phase), wherein the micelle is less than 1 mm, 100 pm, 10 pm, 1 pm, 100 nm, less than 50 nm, less than 25 nm, less than 15 nm or less than 10 nm in size.
- the biosurfactant inhibits the coalescence of the droplets in the continuous phase.
- the emulsion explosive compositions of the subject invention can be applied to a variety of inorganic or organic objects such as, for example, roadway, trail, rock, ore (e.g., coal ore, asbestos ore, iron ore, copper ore, tin ore, nickel ore, gold ore, silver ore, or zinc ore), wood, steel, iron, paint, plastic, paper, silk, glass, cotton, concrete, plaster, clay, stucco, plastic, rubber, or soil.
- the compositions can be applied to objects that reside at a range of temperatures, aquatic environments, or other conditions.
- the composition can be applied to the surface by spraying using, for example, a spray bottle or a pressurized spraying device.
- the composition can also be applied using a cloth or a brush, wherein the composition is rubbed, spread or brushed onto the surface.
- the composition can be applied to the surface by dipping, dunking or submerging the surface into a container having the composition therein.
- the emulsion explosive composition can be applied to an object or surface before initiating an explosion.
- the methods comprise applying a liquid form of the emulsion explosive composition to the surface or object for the period of time before initiating the explosive of at least about 1 sec, about 1 min, about 10 min, about 1 h, about 2 h, about 3 h, about 4 h, about 5 h, about 6 h, about 12 h, about 18 h, about 1 day, about 2 days, about 3 days, about 7 days, about 14 days, about 21 days, about 1 month, about 3 months, or longer.
- the explosion can be initiated using a detonator, such as, for example, electric blasting caps of instantaneous and delay types, blasting caps for use with safety fuses, detonating cord delay connectors, and nonelectric instantaneous and delay blasting caps that use detonating cord, shock tube, or any other replacement for electric legwires.
- a detonator such as, for example, electric blasting caps of instantaneous and delay types, blasting caps for use with safety fuses, detonating cord delay connectors, and nonelectric instantaneous and delay blasting caps that use detonating cord, shock tube, or any other replacement for electric legwires.
- the amount of the emulsion explosive 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 an amount of the object that is being exploded.
- the methods of the subject invention result in at least a 25% increase in the stability of the oxidizer droplets in the composition, preferably at least a 50% increase.
- the emulsion explosive composition according to the subject invention provides enhanced or increased efficiency of explosions and stability of emulsion explosive compositions 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 emulsion explosive composition can be performed on site, including, for example, at a mine or at a roadway. In certain embodiments, the subject emulsion explosive composition can result in a decreased use of chemical surfactants or other potentially harmful chemicals.
- 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. radiobactery, Arthrobacter spp.; Aspergillus spp.; Aureobasidium spp. (e.g., A. pullulans)', Azotobacter (e.g., A. vinelandii, A. chroococcum); Azospirillum spp. (e.g., A. brasiliensisy Bacillus spp.
- Agrobacterium spp. e.g., A. radiobactery, Arthrobacter spp.; Aspergillus spp.
- Aureobasidium spp. e.g., A. pullulans
- Azotobacter e.
- Candida spp. e.g., C. albicans, C. rugosa, C. tropicalis, C. lipolytica, C. torulopsis
- Clostridium e.g., C. butyricum, C. tyrobutyricum, C. acetobutyricum, and C.
- Rhodococcus spp. e.g., R. erythropolisy, Rhodospirillum spp. (e.g., R. riibrumy. Rhizobium spp.; Rhizopus spp.; Saccharomyces spp. (e.g., S. cerevisiae, S. boulardii sequela, S. toruldy Sphingomonas spp. (e.g., .S', paucimobilis); Starmerella spp. (e.g., S.
- the microorganism is a Starmerella spp. yeast and/or Candida spp. yeast, e.g., Starmerella (Candida) bombicola, Candida apicola, Candida batistae, Candida floricola, Candida riodocensis, Candida stellate and/or Candida kuoi.
- the microorganism is Starmerella bombicola, e.g., strain ATCC 22214.
- 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 subject invention provides materials and methods for the production of biomass (e.g., viable cellular material), extracellular metabolites (e.g., small molecules and excreted proteins), residual nutrients and/or intracellular components (e.g., enzymes and other proteins).
- biomass e.g., viable cellular material
- extracellular metabolites e.g., small molecules and excreted proteins
- residual nutrients and/or intracellular components e.g., enzymes and other proteins.
- the microbe growth vessel used according to the subject invention can be any fermenter or cultivation reactor for industrial use.
- the vessel may have functional controls/sensors or may be connected to functional controls/sensors to measure important factors in the cultivation process, such as pH, oxygen, pressure, temperature, humidity, microbial density and/or metabolite concentration.
- 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.
- the method can further comprise supplementing the cultivation with a carbon source.
- the carbon source is typically a carbohydrate, such as glucose, sucrose, lactose, fructose, trehalose, mannose, mannitol, and/or maltose; organic acids such as acetic acid, fumaric acid, citric acid, propionic acid, malic acid, malonic acid, and/or pyruvic acid; alcohols such as ethanol, propanol, butanol, pentanol, hexanol, isobutanol, and/or glycerol; fats and oils such as soybean oil, canola oil, rice bran oil, olive oil, com oil, sesame oil, and/or linseed oil; etc.
- These carbon sources may be used independently or in a combination of two or more.
- growth factors and trace nutrients for microorganisms are included in the medium. This is particularly preferred when growing microbes that are incapable of producing all of the vitamins they require.
- Inorganic nutrients including trace elements such as iron, zinc, copper, manganese, molybdenum and/or cobalt may also be included in the medium.
- sources of vitamins, essential amino acids, and microelements can be included, for example, in the form of flours or meals, such as com flour, or in the form of extracts, such as yeast extract, potato extract, beef extract, soybean extract, banana peel extract, and the like, or in purified forms.
- Amino acids such as, for example, those useful for biosynthesis of proteins, can also be included.
- inorganic salts may also be included.
- 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 may further comprise adding additional acids and/or antimicrobials in the medium before, and/or during the cultivation process.
- Antimicrobial agents or antibiotics are used for protecting the culture against contamination.
- antifoaming agents may also be added to prevent the formation and/or accumulation of foam during submerged cultivation.
- the pH of the mixture should be suitable for the microorganism of interest. Buffers, and pH regulators, such as carbonates and phosphates, may be used to stabilize pH near a preferred value. When metal ions are present in high concentrations, use of a chelating agent in the medium may be necessary.
- the microbes can be grown in planktonic form or as biofilm.
- the vessel may have within it a substrate upon which the microbes can be grown in a biofilm state.
- the system may also have, for example, the capacity to apply stimuli (such as shear stress) that encourages and/or improves the biofilm growth characteristics.
- 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 equipment used in the method and cultivation process is sterile.
- the cultivation equipment such as the reactor/vessel may be separated from, but connected to, a sterilizing unit, e.g., an autoclave.
- the cultivation equipment may also have a sterilizing unit that sterilizes in situ before starting the inoculation.
- Air can be sterilized by methods know in the art.
- the ambient air can pass through at least one filter before being introduced into the vessel.
- the medium may be pasteurized or, optionally, no heat at all added, where the use of low water activity and low pH may be exploited to control undesirable bacterial growth.
- the subject invention further provides a method for producing microbial metabolites such as, for example, biosurfactants, enzymes, proteins, ethanol, lactic acid, beta-glucan, peptides, metabolic intermediates, polyunsaturated fatty acid, and lipids, by cultivating a microbe strain of the subject invention under conditions appropriate for growth and metabolite production; and, optionally, purifying the metabolite.
- microbial metabolites such as, for example, biosurfactants, enzymes, proteins, ethanol, lactic acid, beta-glucan, peptides, metabolic intermediates, polyunsaturated fatty acid, and lipids.
- the metabolite content produced by the method can be, for example, at least 20%, 30%, 40%, 50%, 60%, 70 %, 80 %, or 90%.
- the microbial growth by-product produced by microorganisms of interest may be retained in the microorganisms or secreted into the growth medium.
- the medium may contain compounds that stabilize the activity of microbial growth by-product.
- 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 I0 11 , 1 x 10 8 to 1 x 10 10 , or 1 x 10 9 CFU/ml.
- the method and equipment for cultivation of microorganisms and production of the microbial by-products can be performed in a batch, a quasi-continuous process, or a continuous process.
- 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.
- 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 microbes may be present in or removed from the composition.
- the microbes can be present, with broth in which they were grown, in the microbe-based composition.
- the cells may be present at, for example, a concentration of at least 1 x IO 3 , 1 x 10 4 , 1 x 10 s , 1 x 10 6 , 1 x 10 7 , 1 x 10 8 , 1 x 10 9 , 1 x IO 10 , 1 x 10 n , 1 x 10 12 , 1 x 10 13 or more CFU per milliliter of the composition.
- 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, drying, 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.
- the 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.
- different strains of microbe are grown separately and then mixed together to produce the microbe-based product.
- the microbes can, optionally, be blended with the medium in which they are grown and dried prior to mixing.
- microbe-based products may be used without further stabilization, preservation, and storage.
- direct usage of these microbe-based products preserves a high viability of the microorganisms, reduces the possibility of contamination from foreign agents and undesirable microorganisms, and maintains the activity of the by-products of microbial growth.
- the additives can be, for example, buffers, carriers, other microbe-based compositions produced at the same or different facility, viscosity modifiers, preservatives, nutrients for microbe growth, surfactants, emulsifying agents, lubricants, solubility controlling agents, tracking agents, solvents, biocides, antibiotics, pH adjusting agents, chelators, stabilizers, ultra-violet light resistant agents, other microbes and other suitable additives that are customarily used for such preparations.
- the product can be stored prior to use.
- the storage time is preferably short.
- 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.
- the product is stored at a cool temperature such as, for example, less than 20° C, 15° C, 10° C, or 5° C.
- a biosurfactant composition can typically be stored at ambient temperatures.
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Abstract
The subject invention provides safe, environmentally-friendly compositions and efficient methods for initiating explosions. More specifically, the subject invention provides compositions derived from microorganisms for emulsion explosives, which can also be used for stabilizing emulsion explosive compositions.
Description
EMULSION EXPLOSIVE COMPOSITIONS AND METHODS OF USE
CROSS-REFERENCE TO RELATED APPLICATION
This application claims priority to U.S. Provisional Patent Application No. 63/394,782, filed August 3, 2022, which is incorporated herein by reference in its entirety.
BACKGROUND OF THE INVENTION
Conventional emulsion explosives are water-in-oil emulsions, comprising inorganic oxidizer salt solution droplets (i.e., discontinuous phase) dispersed in a fuel (i.e., continuous phase). The droplets, which constitute a dispersion or emulsion phase, are held in place by a water-in-oil emulsifier provided the emulsified state remains stable. Commonly, an emulsifier or surfactant is used in the emulsion for promoting formation and stability of the oxidizer salt solution droplets.
The detonation velocity characteristic of emulsion explosives makes this class of explosives particularly useful. The useful explosive characteristics are based on the mixing of the emulsion explosive. The continuous phase of fuel surrounds each discontinuous phase droplet of inorganic oxidizer, providing a large interfacial surface area between the two phases.
However, there remain issues with emulsion explosive compositions and methods of use. Emulsion explosives have relatively low densities, which limits the detonation pressure of the explosive. Additionally, the oxidizers used in emulsion explosives can crystallize at ambient air temperatures and must often be kept warmed. If the oxidizer crystallizes, the crystal structure can puncture the oxidizer-containing droplets, causing the droplets to collapse and the crystals to agglomerate. Eventually, the emulsion is destabilized and the usefulness of the emulsion explosive is reduced, possibly to the point in which the emulsion explosive is not detonatable.
Therefore, novel, improved emulsion explosive compositions and methods of use are needed.
BRIEF SUMMARY OF THE INVENTION
The subject invention relates generally to the emulsion explosives. More specifically, the subject invention provides environmentally-friendly compositions and explosion methods, such as, for example, exploding geological formations at mining sites, construction sites, or infrastructure sites. In certain embodiments, the explosion emulsion compositions can be stabilized with the subject compositions and methods.
Advantageously, the compositions and methods of the subject invention increase the effectiveness of emulsion explosives and can decrease the chemical usage, including chemical surfactant, used for emulsion explosive compositions. Accordingly, the subject invention can be useful for reducing the pollution produced during explosions, including during mining operations, creation of roadways, and construction.
In certain embodiments, the subject invention provides compositions comprising components that are derived from microorganisms. In certain embodiments, the composition comprises a microbial biosurfactant. In certain embodiments, the composition comprises one or more biosurfactants, and, optionally, other compounds, such as, for example, water; chemical surfactants; solid fuels, including, for example, aluminum particles, carbonaceous materials (e.g., gilsonite or coal), vegetable grain (e.g., wheat), and sulfur; liquid fuels, including, for example, mineral oil, silicone oil, waxes, paraffin oils, esters (e.g., dioctyl adipate and isodecyl pelargonate), petroleum distillates (e.g., gasoline, kerosene, and diesel), vegetable oils, (e.g., corn oil, cottonseed oil, peanut oil, and soybean oil), aliphatic or aromatic nitro-compounds, and chlorinated hydrocarbons; cross-linkers; oxidizers; inorganic salts, including, for example, ammonium, potassium, and sodium salts of perchlorates and nitrate; emulsifiers, including, for example, sorbitan monooleate, tartaric acid, isopropyl esters of lanolin fatty acids, substituted oxazalines, and polyisobutyl succinic anhydride (PIBSA); or any combination thereof. In certain embodiments, the biosurfactant can produce a foam, liquid, or a semi-solid when applied to a surface.
In certain embodiments, the biosurfactant of the composition is utilized in crude form. The crude form can comprise, in addition to the biosurfactant, fermentation broth in which a biosurfactantproducing microorganism was cultivated, residual microbial cell matter or live or inactive microbial cells, residual nutrients, and/or other microbial growth by-products.
In some embodiments, 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.
In certain specific embodiments, the biosurfactant is a sophorolipid (SLP), including linear SLP, lactonic SLP, acetylated SLP, de-acetylated SLP, salt-form SLP, esterified SLP derivatives, amino acid-SLP conjugates, and other SLP derivatives or isomers that exist in nature and/or are produced synthetically. In preferred embodiments, the SLP is a linear SLP or a derivatized linear SLP.
In certain embodiments, the subject invention provides a method of exploding an object, wherein the method comprises the following steps: a) contacting an emulsion explosive composition according to the subject invention to an object and/or surface; and b) initiating an explosion of the emulsion explosive composition.
In some embodiments, the method reduces the crystallization of oxidizers in the emulsion explosive composition.
In some embodiments, methods of the subject invention comprise contacting an emulsion explosive composition comprising a biosurfactant and, optionally, other components, such as, for example, water; chemical surfactants; solid fuels, including, for example, aluminum particles, carbonaceous materials (e.g., gilsonite or coal), vegetable grain (e.g., wheat), and sulfur; liquid fuels, including, for example, mineral oil, silicone oil, waxes, paraffin oils, esters (e.g., dioctyl adipate and isodecyl pelargonate), petroleum distillates (e.g., gasoline, kerosene, and diesel), vegetable oils, (e.g., com oil, cottonseed oil, peanut oil, and soybean oil), aliphatic or aromatic nitro-compounds, and chlorinated hydrocarbons; cross-linkers; oxidizers; inorganic salts, including, for example, ammonium, potassium, and sodium salts of perchlorates and nitrate; emulsifiers, including, for example, sorbitan monooleate, tartaric acid, isopropyl esters of lanolin fatty acids, substituted oxazalines, and polyisobutyl succinic anhydride (PIBSA); or any combination thereof to an object or surface.
In certain embodiments, the chemical surfactant of the emulsion explosive composition is a detergent, wetting agent, emulsifier, foaming agent, and/or dispersant.
In some embodiments, the method enhances the homogeneity of suspended droplets in the emulsion explosive composition and can increase the surface area of the said droplets. In certain embodiments, the resulting droplets can be less than about 10 cm, about 1 cm, about 1 mm, about 500 pm, about 100 pm, about 10 pm, about 1 pm, about 100 nm, about 10 nm, or about 1 nm in diameter.
In certain embodiments, the effectiveness of emulsion explosive results from the surfactant contacting and stabilizing the discontinuous droplets in the emulsion explosive compositions. This surfactant allows the oxidizer droplets to remain as a liquid, particularly at ambient air temperature.
In certain embodiments, the methods of the subject invention result in at least a 25% increase in the stability of the oxidizer droplets in the composition, preferably at least a 50% increase.
Advantageously, in certain embodiments, the emulsion explosive composition according to the subject invention can be effective at inhibiting the crystallization of oxidizers. Furthermore, the methods of the subject invention do not require complicated equipment or high energy consumption, and production of the composition can be performed on site, for example, at a mine or at a roadway.
DETAILED DESCRIPTION
The subject invention relates generally to emulsion explosive compositions and methods of use. More specifically, the subject invention provides environmentally-friendly compositions and methods for emulsion explosives, such as, for example, for use at mining sites, construction sites, and roadways. In certain embodiments, the emulsion explosive composition can be stabilized with the subject compositions and methods.
Accordingly, the subject invention is useful for improving the efficiency and efficacy of emulsion explosive compositions. Advantageously, the compositions and methods of the subject use safe, environmentally-friendly compositions.
Selected Definitions
As used herein, “applying” a composition or product 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.
As used herein, a “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.
As used herein, 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)) is free of the genes or sequences that flank it in its naturally-occurring state. 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.
In certain embodiments, purified compounds are at least 60% by weight the compound of interest. Preferably, the preparation is at least 75%, more preferably at least 90%, and most preferably at least 98%, by weight the compound of interest. For example, 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.
A “metabolite” refers to any substance produced by metabolism or a substance necessary 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. Examples of metabolites include, but are not limited to, enzymes, acids, solvents, alcohols, proteins, vitamins, minerals, microelements, amino acids, biopolymers and biosurfactants.
Ranges provided herein are understood to be shorthand for all of the values within the range. For example, a range of 1 to 20 is understood to include any number, combination of numbers, or subrange from the group consisting 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 intervening decimal values between the aforementioned integers such as, for example, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, and 1.9. With respect to sub-ranges, “nested sub-ranges” that extend from either end point of the range are specifically contemplated. For example, a nested sub-range of an exemplary range of 1 to 50 may comprise 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 the other direction.
As used herein a “reduction” means a negative alteration, and 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%.
As used herein, “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.
The ability of 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.
Typically, the hydrophilic group of a biosurfactant is a sugar (e.g., a mono-, di-, or polysaccharide) or a peptide, while the hydrophobic group is typically a fatty acid. Thus, there are countless potential variations of 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.
These variations lead to a group of molecules comprising a wide variety of classes, including, for example, glycolipids (e.g., sophorolipids, rhamnolipids, cellobiose lipids, mannosylerythritol lipids and trehalose lipids), lipopeptides (e.g., surfactin, iturin, fengycin, arthrofactin and lichenysin), flavolipids, phospholipids (e.g., cardiolipins), fatty acid ester compounds, 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.
Like chemical surfactants, 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).
The phrases “biosurfactanf ’ 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).
As used herein, the term “sophorolipid,” “sophorolipid molecule,” “SLP” or “SLP molecule” includes all forms, and isomers thereof, of SLP molecules, including, for example, acidic (linear) SLP (ASL) and lactonic SLP (LSL). Further included are mono-acetylated SLP, di-acetylated SLP, esterified SLP, SLP with varying hydrophobic chain lengths, cationic and/or anionic SLP with fatty acid-amino acid complexes attached, esterified SLP, SLP-metal complexes, SLP-salt derivatives (e.g., a sodium salt of a linear SLP), and other, including those that are and/or are not described within in this disclosure.
In certain embodiments, 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 0-glycosidically to 17-L- hydroxyoctadecanoic or 17-L-hydroxy-A9-octadecenoic acid. The hydroxy fatty acid is generally 16 or 18 carbon atoms, and may contain one or more unsaturated bonds. Furthermore, the sophorose residue can be acetylated on the 6- and/or 6’-position(s). The fatty acid carboxyl group can be free (acidic or linear form (General Formula 2)) or internally esterified at the 4"-position (lactonic form (General Formula 1 )). S. bombicola produces a specific enzyme, called S. bombicola lactone esterase, which catalyzes the esterification of linear SLP to produce lactonic SLP.
In preferred embodiments, 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 R1 and R1 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, R2 and R2 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 R3, R3 , R4 and R4 independently represent a hydrogen atom or - COCH3.
The composition utilized according to the subject methods can comprises more than one form of SLP, including linear SLP and lactonic SLP. The SLP can be non-acetylated, mono-acetylated and/or di-acetylated SLP.
In certain specific embodiments, the composition comprises SLP according to General Formula (1 ) (linear SLP) wherein R1 and/or R2 are an acetyl group, and wherein R3 is derived from a stearic, oleic and/or linoleic fatty acid.
SLP are typically produced by yeasts, such as Starmerella spp. yeasts and/or Candida spp. yeasts, e.g., Starmerella (Candida) bombicola, Candida apicola, Candida batistae, Candida floricola,
Candida riodocensis, Candida stellate and/or Candida kuoi. SLP have environmental compatibility, high biodegradability, low toxicity, high selectivity and specific activity in a broad range of temperature, pH and salinity conditions. Additionally, in some embodiments, SLP can be advantageous due to their small micelle size, which can help facilitate the movement of the micelle, and compounds enclosed therein, through nanoscale pores and spaces. In certain embodiments, the micelle size of a 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.
In certain embodiments, the glycolipid is a rhamnolipid. Rhamnolipids comprise a glycosyl head group (i.e., a rhamnose) moiety, and a 3-(hydroxyalkanoyloxy)alkanoic acid (HAA) fatty acid tail, such as, e.g., 3-hydroxydecanoic acid. Two main subtypes of rhamnolipids exist, mono- and dirhamnolipids, which comprise one or two rhamnose moieties, respectively. The HAA moiety can vary in length and degree of branching, depending on, for example, the growth medium and the environmental conditions. The highest accumulation of rhamnolipids (RLP) has been shown by submerged cultivation of Pseudomonas spp., such as P. aeruginosa.
Rhamnolipids according to the subject invention can have the following structure, according to General Formula (3):
wherein m is 2, 1 or 0, n is 1 or 0,
R1 and R2 are, independently of one another, the same or a different organic functional group having 2 to 24, preferably 5 to 13 carbon atoms, in particular a substituted or unsubstituted, branched or unbranched alkyl functional group, which can also be unsaturated, wherein the alkyl functional group is a linear saturated alkyl functional group having 8 to 12 carbon atoms, or is a nonyl or a decyl functional group or a mixture thereof.
Salts of these compounds are also included according to the invention. In the present invention, the term “di-rhamnolipid” is understood to mean compounds of the above formula or the salts thereof in which n is 1. Accordingly, “mono-rhamnolipid” is understood in the present invention to mean compounds of the general formula or the salts thereof in which n is 0. In certain specific embodiments, the composition comprises a mixture of mono- and di-rhamnolipids.
As used herein, “emulsion explosive” refers to a composition comprising two immiscible liquids, such as, for example, oil and water, which is formed by shearing individual droplets (i.e., a discontinuous phase) into a continuous phase. The “continuous phase,” as used herein, is usually a liquid fuel (e.g., oil), while the “discontinuous phase,” as used herein is usually an oxidizer. Emulsion explosive can further comprise a surfactant to stabilize the droplets in the continuous phase.
As used herein, “detonation velocity” is the velocity at which a shock wave travels through a detonated explosive.
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. By 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 a claim to the specified materials or steps “and those that do not materially affect the basic and novel characteristic(s)” of the claimed invention. Use of the term “comprising” contemplates other embodiments that “consist” or “consist essentially of’ the recited component(s).
Unless specifically stated or obvious from context, as used herein, the term "or" is understood to be inclusive. Unless specifically stated or obvious from context, as used herein, the terms “a,” “and” and “the” are understood to be singular or plural.
Unless specifically stated or obvious from context, as used herein, 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 recitation of a listing of chemical groups in any definition of a variable herein includes definitions of that variable as any single group or combination of listed groups. The recitation of an embodiment for a variable or aspect herein includes that embodiment as any single embodiment or in combination with any other embodiments or portions thereof.
All references cited herein are hereby incorporated by reference in their entirety.
Emulsion Explosive Compositions
In certain embodiments, the subject invention provides emulsion explosive compositions comprising components that are derived from microorganisms. In certain embodiments, the emulsion explosive composition comprises a microbial biosurfactant. In certain embodiments, the composition comprises a biosurfactant, and, optionally, water, chemical surfactants, solid fuels, including, for example, aluminum particles, carbonaceous materials (e.g., gilsonite or coal), vegetable grain (e.g., wheat), and sulfur; liquid fuels, including, for example, mineral oil, silicone oil, waxes, paraffin oils, esters (e.g., dioctyl adipate and isodecyl pelargonate), petroleum distillates (e.g., gasoline, kerosene, and diesel), vegetable oils, (e.g., com oil, cottonseed oil, peanut oil, and soybean oil), aliphatic or aromatic nitro-com pounds, and chlorinated hydrocarbons; cross-linkers; oxidizers; inorganic salts, including, for example, ammonium, potassium, and sodium salts of perchlorates and nitrate; emulsifiers, including, for example, sorbitan monooleate, tartaric acid, isopropyl esters of lanolin fatty acids, substituted oxazalines, and polyisobutyl succinic anhydride (PIBSA); or any combination thereof.
In certain embodiments, the emulsion explosive composition comprises a microbe-based product comprising a biosurfactant utilized in crude form. The crude form can comprise, in addition to the biosurfactant, 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 product may be, for example, at least, by weight, 1%, 5%, 10%, 25%, 50%, 75%, or 100% broth. The amount of biomass in the product, by weight, may be, for example, anywhere from 0% to 100% inclusive of all percentages therebetween.
In some embodiments, 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.
In certain specific embodiments, 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. In preferred embodiments, the SLP is a linear SLP or a derivatized linear SLP. In certain embodiments, 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.
In some embodiments, 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 emulsion explosive composition.
In another embodiment, 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).
In some embodiments, 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 the object and/or surface being treated.
In certain embodiments, the chemical surfactant of the emulsion explosive composition is a detergent, wetting agent, emulsifier, foaming agent, and/or dispersant. In some embodiments, the chemical surfactant 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 emulsion explosive composition.
In certain embodiments, the concentrations of the fuel of the continuous phase and liquid oxidizer of the discontinuous phase may be vary depending on the exact components of the phases and any other additives. In certain embodiments, the fuel is about 4 wt% to about 30 wt% or about 5 wt% to about 15 wt% of the emulsion explosive. Preferably, the liquid oxidizer comprises about 70 wt% to about 98 wt% or about 85 wt% to about 95 wt% of the composition. The biosurfactant comprises, preferably, about 0.25 wt% to about 5 wt% of the composition.
In certain embodiments, the solid fuel is aluminum particles, carbonaceous materials (e.g., gilsonite or coal), vegetable grain (e.g., wheat), or sulfur.
In certain embodiments, the liquid fuels are mineral oil, silicone oil, waxes, paraffin oils, esters (e.g., dioctyl adipate and isodecyl pelargonate), petroleum distillates (e.g., gasoline, kerosene, and diesel), vegetable oils, (e.g., com oil, cottonseed oil, peanut oil, and soybean oil), aliphatic or aromatic nitro-compounds, or chlorinated hydrocarbons.
In certain embodiments, the inorganic salts are ammonium, potassium, or sodium salts of perchlorates or nitrate.
In certain embodiments, the emulsifiers are sorbitan monooleate, tartaric acid, isopropyl esters of lanolin fatty acids, substituted oxazalines, or polyisobutyl succinic anhydride (PIBSA).
The emulsion explosive composition can further comprise other additives such as, for example, carriers, other microbe-based compositions, additional biosurfactants, enzymes, catalysts, solvents, organic salts, buffers, chelating agents, acids, lubricants, solubility controlling agents, preservatives, stabilizers, ultra-violet light resistant agents, viscosity modifiers, preservatives, tracking agents, biocides, and other microbes and other ingredients specific for an intended use.
In certain embodiments, chelating agents can be, but are not limited to, ethylenediaminetetraacetic acid (EDTA), nitrilotriacetic acid (NTA), a phosphonate, succimer (DMSA), diethylenetriaminepentaacetate (DTPA), A'-acetylcysteine, n- hydroxyethylethylenediaminetriacetic acid (HEDTA), organic acids with more than one coordination group (e.g., rubeanic acid), STPP (sodiumtripolyphosphate, Na5P3O10), trisodium phosphate (TSP), water, carbohydrates, organic acids with more than one coordination group (e.g., citric acid), lipids, steroids, amino acids or related compounds (e.g., glutathione), peptides, phosphates, nucleotides, tetrapyrrols, ferrioxamines, ionophores, orphenolics, sodium citrate, sodium gluconate, ethylenediamine disuccinic acid (EDDS), iminodisuccinic acid (IDS), L-glutamic acid diacetic Acid (GLDA), GLDA-Na4, methyl glycindiacetic acid (MGDA), polyaspartic acid (PASA), hemoglobin, chlorophyll, lipophilic P-diketone, and (14,16)-hentriacontanedione, 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), trimethyl glycine (TMG), Tiron, or any combination thereof.
In certain embodiments, the emulsion explosives of embodiments of the present invention may be formulated using conventional procedures known in the art. In certain embodiments, the liquid oxidizer, fuel, and biosurfactant solution are approximately equal in temperature when combined. The temperature is preferably in a range at which the oxidizer will remain in a liquid state. Accordingly, the selected temperature will be dependent upon the particular oxidizer selected. The resulting mixture can be stirred or otherwise mixed vigorously to produce an emulsion of the liquid oxidizer in a continuous liquid fuel phase. Stirring should be continued until the formulation is uniform. The formulation process also can be accomplished in a continuous manner as is known in the art. Various modifications to the above-described technique are possible. In certain embodiments, the biosurfactant can be dissolved in the liquid organic fuel prior to combining the organic fuel with the liquid oxidizer to form an emulsion. This method allows the emulsion to form quickly and without excessive agitation. However, the emulsifier may be added separately as a third component if desired, may be combined with the liquid oxidizer, or may be combined with the liquid oxidizer and the liquid organic fuel independently before the mixing of the liquid oxidizer and the liquid organic fuel.
Methods of Use of the Emulsion Explosives
In certain embodiments, the subject invention provides a method for initiating explosions at various sites, including mining sites, tunneling sites, quarrying sites, construction sites, and roadways. In certain specific embodiments, the emulsion explosives can be used to displace geological formations and existing human-derived constructions, such as, for example, infrastructure, roadways, buildings,
and bridges. The described elements of the subject invention are not an exhaustive examination of all applications.
In certain embodiments, the mining site can be a coal mine, iron ore mine (e.g., taconite), copper mine, copper-nickel mine, tin mine, nickel mine, gold mine, silver mine, molybdenum mine, aluminum mine (e.g., bauxite mine, kyanite mine), lead-zine mine, tungsten mine, phosphate mine, potash mine, mica mine, bentonite mine, or zinc mine. The mine can be an underground mine, surface mine, placer mine or in situ mine. In certain embodiments, the roadway can be asphalt, tar and chip, earthen roads (e.g., dirt and gravel), or concrete. The road surfaces can be intended for use for semi-trucks or other industrial machinery, including for example, excavator or dump trucks, or for use as an automobile, bicycle, or motorcycle racetrack. In certain embodiments, the construction site can be the site of producing a commercial building, including, for example, an office or warehouse; residential building, including, for example, a single family or multifamily home; an industrial facility, including for example, a factory; or an infrastructure project, such as, for example, a roadway, bridge, waterway, or sewer. In certain embodiments, the quarrying site can extract chalk, clay, cinder, coal, sand, gravel, coquina, diabase, gabbro, granite, gritstone, gypsum, limestone, marble, ores, phosphate rock, quartz, sandstone, slate, travertine, or any combination thereof.
In certain embodiments, the microbe-containing and/or biosurfactant-containing composition can improve the stability of the oxidizer droplets (i.e., inhibit or reduce crystallization of the oxidizer) within the continuous phase (e.g., fuel), particularly at a range of temperatures including less about 150°C, about 125°C, about 100°C, about 90°C, about 80°C, about 75°C, about 70°C, about 65°C, about 60°C, about 55°C, about 50°C, about 45°C, about 40°C, about 35°C, about 32°C, about 30°C, or lower.
In certain embodiments, the microbe-containing and/or biosurfactant-containing composition can stabilize droplets suspended (i.e., discontinuous phase) in a liquid continuous phase. This also allows for an increase in the total surface area of all the droplets within the continuous phase. This increase of the surface area of the droplets allows the for a more efficient explosion, yielding a greater detonation velocity. In certain embodiments, the detonation velocity can be about 18,000 to about 25,000 feet per second (5490 m/s to 7620 m/s), which is determined by the amount of surface area of the discontinuous phase. The continuous phase of fuel surrounds each discontinuous phase droplet of inorganic oxidizer, providing a large interfacial surface area between the two phases.
In certain embodiments, the composition can reduce the effects of cool temperature or the presence of water on the effectiveness of the emulsion explosives, particularly during blasting.
In certain embodiments, the microbe-containing and/or biosurfactant-containing composition has increased longevity versus traditional emulsion explosive compositions due to its reduced formation of crystals in the oxidizer droplets. In certain embodiments, the emulsion explosive composition according to the subject invention is effective due to amphiphiles-mediated droplet stability in a
continuous phase. In some embodiments, the sophorolipid or other biosurfactant serves as a means for facilitating the strength of interfacial film in the composition. The interfacial film forms between the two immiscible phases of the composition: the continuous phase and the discontinuous phase. For example, in some embodiments, a sophorolipid will form a micelle containing the oxidizer droplet (i.e., discontinuous phase), wherein the micelle is less than 1 mm, 100 pm, 10 pm, 1 pm, 100 nm, less than 50 nm, less than 25 nm, less than 15 nm or less than 10 nm in size. In certain embodiments, the biosurfactant inhibits the coalescence of the droplets in the continuous phase.
The emulsion explosive compositions of the subject invention can be applied to a variety of inorganic or organic objects such as, for example, roadway, trail, rock, ore (e.g., coal ore, asbestos ore, iron ore, copper ore, tin ore, nickel ore, gold ore, silver ore, or zinc ore), wood, steel, iron, paint, plastic, paper, silk, glass, cotton, concrete, plaster, clay, stucco, plastic, rubber, or soil. The compositions can be applied to objects that reside at a range of temperatures, aquatic environments, or other conditions.
The composition can be applied to the surface by spraying using, for example, a spray bottle or a pressurized spraying device. The composition can also be applied using a cloth or a brush, wherein the composition is rubbed, spread or brushed onto the surface. Furthermore, the composition can be applied to the surface by dipping, dunking or submerging the surface into a container having the composition therein.
In certain embodiments, the emulsion explosive composition can be applied to an object or surface before initiating an explosion. In certain embodiments, the methods comprise applying a liquid form of the emulsion explosive composition to the surface or object for the period of time before initiating the explosive of at least about 1 sec, about 1 min, about 10 min, about 1 h, about 2 h, about 3 h, about 4 h, about 5 h, about 6 h, about 12 h, about 18 h, about 1 day, about 2 days, about 3 days, about 7 days, about 14 days, about 21 days, about 1 month, about 3 months, or longer. In certain embodiments, the explosion can be initiated using a detonator, such as, for example, electric blasting caps of instantaneous and delay types, blasting caps for use with safety fuses, detonating cord delay connectors, and nonelectric instantaneous and delay blasting caps that use detonating cord, shock tube, or any other replacement for electric legwires.
In certain embodiments, the amount of the emulsion explosive 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 an amount of the object that is being exploded.
In certain embodiments, the methods of the subject invention result in at least a 25% increase in the stability of the oxidizer droplets in the composition, preferably at least a 50% increase.
Advantageously, in certain embodiments, the emulsion explosive composition according to the subject invention provides enhanced or increased efficiency of explosions and stability of emulsion explosive compositions 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 emulsion explosive composition can be performed on site, including, for example, at a mine or at a roadway. In certain embodiments, the subject emulsion explosive composition can result in a decreased use of chemical surfactants or other potentially harmful chemicals.
Production of Microbe-Based Products
In certain embodiments, 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.
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. As used herein, “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.
In certain embodiments, the microbes are capable of producing amphiphilic molecules, enzymes, proteins and/or biopolymers. Microbial biosurfactants, in particular, are produced by a variety of microorganisms such as bacteria, fungi, and yeasts, including, for example, Agrobacterium spp. (e.g., A. radiobactery, Arthrobacter spp.; Aspergillus spp.; Aureobasidium spp. (e.g., A. pullulans)', Azotobacter (e.g., A. vinelandii, A. chroococcum); Azospirillum spp. (e.g., A. brasiliensisy Bacillus spp. (e.g., B. subtilis, B. amyloliquefaciens, B. pumillus, B. cereus, B. licheniformis, B. firmus, B. laterosporus, B. megateriuniy, Blakeslea,' Candida spp. (e.g., C. albicans, C. rugosa, C. tropicalis, C. lipolytica, C. torulopsis),' Clostridium (e.g., C. butyricum, C. tyrobutyricum, C. acetobutyricum, and 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., P. anomala, P. guilliermondii, P. occidentalis, P. kudriavzeviiy Phycomyces spp.; Phythium spp.; Pseudomonas spp. (e.g., P. aeruginosa, P. chlororaphis, P. putida, P.florescens, P.fragi, P. syringae);
Pseudozyma spp. (e.g., P. aphidis),' Ralslonia spp. (e.g., R. eulropha); Rhodococcus spp. (e.g., R. erythropolisy, Rhodospirillum spp. (e.g., R. riibrumy. Rhizobium spp.; Rhizopus spp.; Saccharomyces spp. (e.g., S. cerevisiae, S. boulardii sequela, S. toruldy Sphingomonas spp. (e.g., .S', paucimobilis); Starmerella spp. (e.g., S. bombicoldy, Thraustochytrium spp.; Torulopsis spp.; Ustilago spp. (e.g., U. maydis Wickerhamomyces spp. (e.g., W. anomalusy, Williopsis spp.; and/or Zygosaccharomyces spp. (e.g., Z. bailii).
In preferred embodiments, the microorganism is a Starmerella spp. yeast and/or Candida spp. yeast, e.g., Starmerella (Candida) bombicola, Candida apicola, Candida batistae, Candida floricola, Candida riodocensis, Candida stellate and/or Candida kuoi. In a specific embodiment, the microorganism is Starmerella bombicola, e.g., strain ATCC 22214.
As used herein “fermentation” refers to cultivation or growth of cells under controlled conditions. The growth could be aerobic or anaerobic. In preferred embodiments, the microorganisms are grown using SSF and/or modified versions thereof.
In one embodiment, the subject invention provides materials and methods for the production of biomass (e.g., viable cellular material), extracellular metabolites (e.g., small molecules and excreted proteins), residual nutrients and/or intracellular components (e.g., enzymes and other proteins).
The microbe growth vessel used according to the subject invention can be any fermenter or cultivation reactor for industrial use. In one embodiment, the vessel may have functional controls/sensors or may be connected to functional controls/sensors to measure important factors in the cultivation process, such as pH, oxygen, pressure, temperature, humidity, microbial density and/or metabolite concentration.
In a further embodiment, the vessel may also be able to monitor the growth of microorganisms inside the vessel (e.g., measurement of cell number and growth phases). Alternatively, 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.
In one embodiment, 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. In the case of submerged fermentation, 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.
The method can further comprise supplementing the cultivation with a carbon source. The carbon source is typically a carbohydrate, such as glucose, sucrose, lactose, fructose, trehalose, mannose, mannitol, and/or maltose; organic acids such as acetic acid, fumaric acid, citric acid, propionic acid, malic acid, malonic acid, and/or pyruvic acid; alcohols such as ethanol, propanol, butanol, pentanol, hexanol, isobutanol, and/or glycerol; fats and oils such as soybean oil, canola oil, rice bran oil, olive oil, com oil, sesame oil, and/or linseed oil; etc. These carbon sources may be used independently or in a combination of two or more.
In one embodiment, growth factors and trace nutrients for microorganisms are included in the medium. This is particularly preferred when growing microbes that are incapable of producing all of the vitamins they require. Inorganic nutrients, including trace elements such as iron, zinc, copper, manganese, molybdenum and/or cobalt may also be included in the medium. Furthermore, sources of vitamins, essential amino acids, and microelements can be included, for example, in the form of flours or meals, such as com flour, or in the form of extracts, such as yeast extract, potato extract, beef extract, soybean extract, banana peel extract, and the like, or in purified forms. Amino acids such as, for example, those useful for biosynthesis of proteins, can also be included.
In one embodiment, inorganic salts may also be included. 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.
In some embodiments, the method for cultivation may further comprise adding additional acids and/or antimicrobials in the medium before, and/or during the cultivation process. Antimicrobial agents or antibiotics are used for protecting the culture against contamination.
Additionally, antifoaming agents may also be added to prevent the formation and/or accumulation of foam during submerged cultivation.
The pH of the mixture should be suitable for the microorganism of interest. Buffers, and pH regulators, such as carbonates and phosphates, may be used to stabilize pH near a preferred value. When metal ions are present in high concentrations, use of a chelating agent in the medium may be necessary.
The microbes can be grown in planktonic form or as biofilm. In the case of biofilm, the vessel may have within it a substrate upon which the microbes can be grown in a biofilm state. The system may also have, for example, the capacity to apply stimuli (such as shear stress) that encourages and/or improves the biofilm growth characteristics.
In one embodiment, 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. In a further embodiment, the cultivation may be carried out continuously at a constant temperature. In another embodiment, the cultivation may be subject to changing temperatures.
In one embodiment, the equipment used in the method and cultivation process is sterile. The cultivation equipment such as the reactor/vessel may be separated from, but connected to, a sterilizing unit, e.g., an autoclave. The cultivation equipment may also have a sterilizing unit that sterilizes in situ before starting the inoculation. Air can be sterilized by methods know in the art. For example, the ambient air can pass through at least one filter before being introduced into the vessel. In other embodiments, the medium may be pasteurized or, optionally, no heat at all added, where the use of low water activity and low pH may be exploited to control undesirable bacterial growth.
In one embodiment, the subject invention further provides a method for producing microbial metabolites such as, for example, biosurfactants, enzymes, proteins, ethanol, lactic acid, beta-glucan, peptides, metabolic intermediates, polyunsaturated fatty acid, and lipids, by cultivating a microbe strain of the subject invention under conditions appropriate for growth and metabolite production; and, optionally, purifying the metabolite. The metabolite content produced by the method can be, for example, at least 20%, 30%, 40%, 50%, 60%, 70 %, 80 %, or 90%.
The microbial growth by-product produced by microorganisms of interest may be retained in the microorganisms or secreted into the growth medium. The medium may contain compounds that stabilize the activity of microbial growth by-product.
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 106 to 1 x 1012, 1 x 107 to 1 x I011, 1 x 108to 1 x 1010, or 1 x 109 CFU/ml.
The method and equipment for cultivation of microorganisms and production of the microbial by-products can be performed in a batch, a quasi-continuous process, or a continuous process.
In one embodiment, 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). In this batch procedure, an entirely new batch is initiated upon harvesting of the first batch.
In another embodiment, only a portion of the fermentation product is removed at any one time. In this embodiment, 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.
Advantageously, 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.
In certain embodiments, 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. Thus, 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 microbes may be present in or removed from the composition. The microbes can be present, with broth in which they were grown, in the microbe-based composition. The cells may be present at, for example, a concentration of at least 1 x IO3, 1 x 104, 1 x 10s, 1 x 106, 1 x 107, 1 x 108, 1 x 109, 1 x IO10, 1 x 10n , 1 x 1012, 1 x 1013 or more CFU per milliliter of the composition.
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. Alternatively, 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, drying, 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.
The 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.
In one embodiment, different strains of microbe are grown separately and then mixed together to produce the microbe-based product. The microbes can, optionally, be blended with the medium in which they are grown and dried prior to mixing.
The microbe-based products may be used without further stabilization, preservation, and storage. Advantageously, direct usage of these microbe-based products preserves a high viability of the microorganisms, reduces the possibility of contamination from foreign agents and undesirable microorganisms, and maintains the activity of the by-products of microbial growth.
Upon harvesting the microbe-based composition from the growth vessels, further components can be added as the harvested product is placed into containers or otherwise transported for use. The additives can be, for example, buffers, carriers, other microbe-based compositions produced at the same or different facility, viscosity modifiers, preservatives, nutrients for microbe growth, surfactants, emulsifying agents, lubricants, solubility controlling agents, tracking agents, solvents, biocides, antibiotics, pH adjusting agents, chelators, stabilizers, ultra-violet light resistant agents, other microbes and other suitable additives that are customarily used for such preparations.
Optionally, the product can 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 live cells are present in the product, the product is stored at a cool temperature such as, for example, less than 20° C, 15° C, 10° C, or 5° C. On the other hand, a biosurfactant composition can typically be stored at ambient temperatures.
Claims
1 . A method for initiating an explosion, the method comprising: a) contacting an emulsion explosive composition comprising a biosurfactant to an object and/or surface; and b) initiating an explosion of the emulsion explosive composition.
2. The method of claim 1, wherein the object or surface is at a mining site, quarrying site, tunneling site, construction site, or roadway.
3. The method of claim 2, wherein the mining site is a coal mine, iron ore mine, copper mine, copper-nickel mine, tin mine, nickel mine, gold mine, silver mine, molybdenum mine, aluminum mine, lead-zinc mine, tungsten mine, phosphate mine, potash mine, mica mine, bentonite mine, or zinc mine.
4. The method of claim 2, wherein the quarrying site contains chalk, clay, cinder, coal, sand, gravel, coquina, diabase, gabbro, granite, gritstone, gypsum, limestone, marble, ores, phosphate rock, quartz, sandstone, slate, travertine, or any combination thereof.
5. The method of claim 1 , wherein the composition further comprises a chemical surfactant, water, solid fuel, liquid fuel, cross-linker, oxidizer, inorganic salt, emulsifier, or any combination thereof.
6. The method of claim 5, wherein the inorganic salt is an ammonium, potassium, or sodium salt.
7. The method of claim 6, wherein the ammonium salt is ammonium nitrate.
8. The method of claim 1, wherein the emulsion explosive composition is in liquid form, and wherein the contacting step comprises spraying or pouring the composition on the object or surface.
9. The method of claim 1 , wherein the biosurfactant is a glycolipid and/or a yeast culture comprising a glycolipid.
10. The method of claim 9, wherein the yeast culture is a Starmerella sp. and/or a Candida sp. yeast.
1 1. The method of claim 9, wherein the glycolipid is a sophorolipid, mannosylerythritol lipid, trehalose lipid, rhamnolipid, or any combination thereof.
12. The method of claim 11, wherein the sophorolipid is a linear sophorolipid or a lactonic sophorolipid.
13. A method of stabilizing an emulsion explosive composition, the method comprising contacting a biosurfactant to an oxidizer and a fuel.
14. The method of claim 13, wherein the emulsion explosive composition is stabilized by one or a combination of the following: a) inhibiting crystallization of the oxidizer; or b) inhibiting coalescence of the oxidizer.
15. An emulsion explosive composition comprising a biosurfactant and/or a yeast culture comprising a biosurfactant, and one or more traditional emulsion explosive components.
16. The composition of claim 15, wherein the biosurfactant is a rhamnolipid, sophorolipid, cellobiose lipid, mannosylerythritol lipid, trehalose lipid, lipopeptide, flavolipid, phospholipid, fatty acid ester compound, fatty acid ether compound, or any combination thereof.
17. The composition of claim 15, wherein the yeast culture is a Starmerella sp. and/or a Candida sp. yeast.
18. The composition of claim 16, wherein the sophorolipid is a linear sophorolipid or a lactonic sophorolipid.
19. The composition of claim 15, wherein the emulsion explosive components are selected from one or a combination of the following: a) chemical surfactant; b) water; c) solid fuel; d) liquid fuel; e) cross-linker; f oxidizer;
g) inorganic salt; or h) emulsifier.
20. The composition of claim 19, wherein the inorganic salt is an ammonium, potassium, or sodium salt.
21 . The composition of claim 20, wherein the ammonium salt is ammonium nitrate.
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