CA2396691A1 - Enhanced production of lipids containing polyenoic fatty acids by high density cultures of eukaryotic microbes in fermentors - Google Patents
Enhanced production of lipids containing polyenoic fatty acids by high density cultures of eukaryotic microbes in fermentors Download PDFInfo
- Publication number
- CA2396691A1 CA2396691A1 CA002396691A CA2396691A CA2396691A1 CA 2396691 A1 CA2396691 A1 CA 2396691A1 CA 002396691 A CA002396691 A CA 002396691A CA 2396691 A CA2396691 A CA 2396691A CA 2396691 A1 CA2396691 A1 CA 2396691A1
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- CA
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- Prior art keywords
- sources
- microorganisms
- lipids
- cndot
- fermentation medium
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
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- 150000002632 lipids Chemical class 0.000 title claims abstract 73
- 235000014113 dietary fatty acids Nutrition 0.000 title claims abstract 11
- 229930195729 fatty acid Natural products 0.000 title claims abstract 11
- 239000000194 fatty acid Substances 0.000 title claims abstract 11
- 150000004665 fatty acids Chemical class 0.000 title claims abstract 11
- 238000004519 manufacturing process Methods 0.000 title claims 10
- 238000000034 method Methods 0.000 claims abstract 144
- 244000005700 microbiome Species 0.000 claims abstract 76
- 230000000813 microbial effect Effects 0.000 claims abstract 22
- 238000000855 fermentation Methods 0.000 claims 44
- 230000004151 fermentation Effects 0.000 claims 44
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims 38
- 239000002609 medium Substances 0.000 claims 36
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims 33
- 229910052760 oxygen Inorganic materials 0.000 claims 33
- 239000001301 oxygen Substances 0.000 claims 33
- 239000000203 mixture Substances 0.000 claims 24
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims 21
- 229910052799 carbon Inorganic materials 0.000 claims 21
- MBMBGCFOFBJSGT-KUBAVDMBSA-N all-cis-docosa-4,7,10,13,16,19-hexaenoic acid Chemical compound CC\C=C/C\C=C/C\C=C/C\C=C/C\C=C/C\C=C/CCC(O)=O MBMBGCFOFBJSGT-KUBAVDMBSA-N 0.000 claims 20
- 235000015097 nutrients Nutrition 0.000 claims 20
- 229910052757 nitrogen Inorganic materials 0.000 claims 19
- 239000002028 Biomass Substances 0.000 claims 15
- 229910052751 metal Inorganic materials 0.000 claims 15
- 239000002184 metal Substances 0.000 claims 15
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims 11
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 claims 10
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims 10
- PXHVJJICTQNCMI-UHFFFAOYSA-N Nickel Chemical compound [Ni] PXHVJJICTQNCMI-UHFFFAOYSA-N 0.000 claims 10
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims 10
- 235000020669 docosahexaenoic acid Nutrition 0.000 claims 10
- 229940090949 docosahexaenoic acid Drugs 0.000 claims 10
- 229910021654 trace metal Inorganic materials 0.000 claims 10
- 241000894006 Bacteria Species 0.000 claims 9
- 241000195493 Cryptophyta Species 0.000 claims 9
- 241000233866 Fungi Species 0.000 claims 9
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims 7
- 241001466451 Stramenopiles Species 0.000 claims 6
- 230000015572 biosynthetic process Effects 0.000 claims 6
- 238000005119 centrifugation Methods 0.000 claims 6
- 230000005484 gravity Effects 0.000 claims 6
- 235000020660 omega-3 fatty acid Nutrition 0.000 claims 6
- 235000020665 omega-6 fatty acid Nutrition 0.000 claims 6
- 238000011084 recovery Methods 0.000 claims 6
- 238000000926 separation method Methods 0.000 claims 6
- 238000003786 synthesis reaction Methods 0.000 claims 6
- GHOKWGTUZJEAQD-ZETCQYMHSA-N (D)-(+)-Pantothenic acid Chemical compound OCC(C)(C)[C@@H](O)C(=O)NCCC(O)=O GHOKWGTUZJEAQD-ZETCQYMHSA-N 0.000 claims 5
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 claims 5
- 229910021580 Cobalt(II) chloride Inorganic materials 0.000 claims 5
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 claims 5
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 claims 5
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 claims 5
- PWHULOQIROXLJO-UHFFFAOYSA-N Manganese Chemical compound [Mn] PWHULOQIROXLJO-UHFFFAOYSA-N 0.000 claims 5
- 229910021380 Manganese Chloride Inorganic materials 0.000 claims 5
- GLFNIEUTAYBVOC-UHFFFAOYSA-L Manganese chloride Chemical compound Cl[Mn]Cl GLFNIEUTAYBVOC-UHFFFAOYSA-L 0.000 claims 5
- ZOKXTWBITQBERF-UHFFFAOYSA-N Molybdenum Chemical compound [Mo] ZOKXTWBITQBERF-UHFFFAOYSA-N 0.000 claims 5
- 229910004619 Na2MoO4 Inorganic materials 0.000 claims 5
- 239000007832 Na2SO4 Substances 0.000 claims 5
- 229910019142 PO4 Inorganic materials 0.000 claims 5
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 claims 5
- 241000233671 Schizochytrium Species 0.000 claims 5
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 claims 5
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 claims 5
- JZRWCGZRTZMZEH-UHFFFAOYSA-N Thiamine Natural products CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N JZRWCGZRTZMZEH-UHFFFAOYSA-N 0.000 claims 5
- 241001467333 Thraustochytriaceae Species 0.000 claims 5
- 229930003779 Vitamin B12 Natural products 0.000 claims 5
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 claims 5
- 150000003863 ammonium salts Chemical class 0.000 claims 5
- 239000011575 calcium Substances 0.000 claims 5
- 229910052791 calcium Inorganic materials 0.000 claims 5
- 150000003841 chloride salts Chemical class 0.000 claims 5
- 229910017052 cobalt Inorganic materials 0.000 claims 5
- 239000010941 cobalt Substances 0.000 claims 5
- GUTLYIVDDKVIGB-UHFFFAOYSA-N cobalt atom Chemical compound [Co] GUTLYIVDDKVIGB-UHFFFAOYSA-N 0.000 claims 5
- AGVAZMGAQJOSFJ-WZHZPDAFSA-M cobalt(2+);[(2r,3s,4r,5s)-5-(5,6-dimethylbenzimidazol-1-yl)-4-hydroxy-2-(hydroxymethyl)oxolan-3-yl] [(2r)-1-[3-[(1r,2r,3r,4z,7s,9z,12s,13s,14z,17s,18s,19r)-2,13,18-tris(2-amino-2-oxoethyl)-7,12,17-tris(3-amino-3-oxopropyl)-3,5,8,8,13,15,18,19-octamethyl-2 Chemical compound [Co+2].N#[C-].[N-]([C@@H]1[C@H](CC(N)=O)[C@@]2(C)CCC(=O)NC[C@@H](C)OP(O)(=O)O[C@H]3[C@H]([C@H](O[C@@H]3CO)N3C4=CC(C)=C(C)C=C4N=C3)O)\C2=C(C)/C([C@H](C\2(C)C)CCC(N)=O)=N/C/2=C\C([C@H]([C@@]/2(CC(N)=O)C)CCC(N)=O)=N\C\2=C(C)/C2=N[C@]1(C)[C@@](C)(CC(N)=O)[C@@H]2CCC(N)=O AGVAZMGAQJOSFJ-WZHZPDAFSA-M 0.000 claims 5
- 229910052802 copper Inorganic materials 0.000 claims 5
- 239000010949 copper Substances 0.000 claims 5
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 claims 5
- 229910000366 copper(II) sulfate Inorganic materials 0.000 claims 5
- XLYOFNOQVPJJNP-ZSJDYOACSA-N heavy water Substances [2H]O[2H] XLYOFNOQVPJJNP-ZSJDYOACSA-N 0.000 claims 5
- 229910052742 iron Inorganic materials 0.000 claims 5
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 claims 5
- 229910000359 iron(II) sulfate Inorganic materials 0.000 claims 5
- 239000011777 magnesium Substances 0.000 claims 5
- 229910052749 magnesium Inorganic materials 0.000 claims 5
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims 5
- 235000019341 magnesium sulphate Nutrition 0.000 claims 5
- 229910052748 manganese Inorganic materials 0.000 claims 5
- 239000011572 manganese Substances 0.000 claims 5
- 239000011565 manganese chloride Substances 0.000 claims 5
- 235000002867 manganese chloride Nutrition 0.000 claims 5
- 150000002739 metals Chemical class 0.000 claims 5
- 229910052750 molybdenum Inorganic materials 0.000 claims 5
- 239000011733 molybdenum Substances 0.000 claims 5
- 229910052759 nickel Inorganic materials 0.000 claims 5
- LGQLOGILCSXPEA-UHFFFAOYSA-L nickel sulfate Chemical compound [Ni+2].[O-]S([O-])(=O)=O LGQLOGILCSXPEA-UHFFFAOYSA-L 0.000 claims 5
- 229910000363 nickel(II) sulfate Inorganic materials 0.000 claims 5
- 229940012843 omega-3 fatty acid Drugs 0.000 claims 5
- 229940033080 omega-6 fatty acid Drugs 0.000 claims 5
- 229940014662 pantothenate Drugs 0.000 claims 5
- 235000019161 pantothenic acid Nutrition 0.000 claims 5
- 239000011713 pantothenic acid Substances 0.000 claims 5
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims 5
- 239000010452 phosphate Substances 0.000 claims 5
- 239000011591 potassium Substances 0.000 claims 5
- 229910052700 potassium Inorganic materials 0.000 claims 5
- 229910052939 potassium sulfate Inorganic materials 0.000 claims 5
- OTYBMLCTZGSZBG-UHFFFAOYSA-L potassium sulfate Chemical compound [K+].[K+].[O-]S([O-])(=O)=O OTYBMLCTZGSZBG-UHFFFAOYSA-L 0.000 claims 5
- 239000000377 silicon dioxide Substances 0.000 claims 5
- 239000011734 sodium Substances 0.000 claims 5
- 229910052708 sodium Inorganic materials 0.000 claims 5
- 235000015393 sodium molybdate Nutrition 0.000 claims 5
- 239000011684 sodium molybdate Substances 0.000 claims 5
- TVXXNOYZHKPKGW-UHFFFAOYSA-N sodium molybdate (anhydrous) Chemical compound [Na+].[Na+].[O-][Mo]([O-])(=O)=O TVXXNOYZHKPKGW-UHFFFAOYSA-N 0.000 claims 5
- 229910052938 sodium sulfate Inorganic materials 0.000 claims 5
- 235000011152 sodium sulphate Nutrition 0.000 claims 5
- KYMBYSLLVAOCFI-UHFFFAOYSA-N thiamine Chemical compound CC1=C(CCO)SCN1CC1=CN=C(C)N=C1N KYMBYSLLVAOCFI-UHFFFAOYSA-N 0.000 claims 5
- 229960003495 thiamine Drugs 0.000 claims 5
- 235000019157 thiamine Nutrition 0.000 claims 5
- 239000011721 thiamine Substances 0.000 claims 5
- 229940088594 vitamin Drugs 0.000 claims 5
- 229930003231 vitamin Natural products 0.000 claims 5
- 235000013343 vitamin Nutrition 0.000 claims 5
- 239000011782 vitamin Substances 0.000 claims 5
- 235000019163 vitamin B12 Nutrition 0.000 claims 5
- 239000011715 vitamin B12 Substances 0.000 claims 5
- 150000003722 vitamin derivatives Chemical class 0.000 claims 5
- 229910052725 zinc Inorganic materials 0.000 claims 5
- 239000011701 zinc Substances 0.000 claims 5
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 claims 5
- 229910000368 zinc sulfate Inorganic materials 0.000 claims 5
- 239000011686 zinc sulphate Substances 0.000 claims 5
- 235000009529 zinc sulphate Nutrition 0.000 claims 5
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 claims 4
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims 4
- 108010030975 Polyketide Synthases Proteins 0.000 claims 4
- 241000233675 Thraustochytrium Species 0.000 claims 4
- 239000000908 ammonium hydroxide Substances 0.000 claims 4
- 239000001110 calcium chloride Substances 0.000 claims 4
- 235000011148 calcium chloride Nutrition 0.000 claims 4
- 229910001628 calcium chloride Inorganic materials 0.000 claims 4
- 150000001720 carbohydrates Chemical class 0.000 claims 4
- 239000000839 emulsion Substances 0.000 claims 3
- 230000003698 anagen phase Effects 0.000 claims 2
- 239000003795 chemical substances by application Substances 0.000 claims 2
- 238000001704 evaporation Methods 0.000 claims 2
- 239000001963 growth medium Substances 0.000 claims 2
- 238000010923 batch production Methods 0.000 claims 1
- 238000012258 culturing Methods 0.000 claims 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
- C12P7/6409—Fatty acids
- C12P7/6427—Polyunsaturated fatty acids [PUFA], i.e. having two or more double bonds in their backbone
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/80—Feeding-stuffs specially adapted for particular animals for aquatic animals, e.g. fish, crustaceans or molluscs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23D—EDIBLE OILS OR FATS, e.g. MARGARINES, SHORTENINGS, COOKING OILS
- A23D9/00—Other edible oils or fats, e.g. shortenings, cooking oils
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/158—Fatty acids; Fats; Products containing oils or fats
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/065—Microorganisms
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/02—Nutrients, e.g. vitamins, minerals
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B1/00—Production of fats or fatty oils from raw materials
- C11B1/10—Production of fats or fatty oils from raw materials by extracting
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/12—Unicellular algae; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
- C12P7/6409—Fatty acids
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
- C12P7/6409—Fatty acids
- C12P7/6427—Polyunsaturated fatty acids [PUFA], i.e. having two or more double bonds in their backbone
- C12P7/6434—Docosahexenoic acids [DHA]
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
- C12P7/6436—Fatty acid esters
- C12P7/6445—Glycerides
- C12P7/6472—Glycerides containing polyunsaturated fatty acid [PUFA] residues, i.e. having two or more double bonds in their backbone
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S435/00—Chemistry: molecular biology and microbiology
- Y10S435/8215—Microorganisms
- Y10S435/911—Microorganisms using fungi
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- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Polymers & Plastics (AREA)
- Genetics & Genomics (AREA)
- Microbiology (AREA)
- General Health & Medical Sciences (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Food Science & Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Animal Husbandry (AREA)
- Medicinal Chemistry (AREA)
- Biomedical Technology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Nutrition Science (AREA)
- Insects & Arthropods (AREA)
- Birds (AREA)
- Marine Sciences & Fisheries (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Animal Behavior & Ethology (AREA)
- Diabetes (AREA)
- Hematology (AREA)
- Obesity (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Pharmacology & Pharmacy (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The present invention provides a process for growing eukaryotic microorganisms, which are capable of producing lipids, in particular, lipids containing polyenoic fatty acids. The present invention also provides a process for producing eukaryotic microbial lipids.
Claims (131)
1. A process for producing lipids containing polyenoic fatty acids from eukaryotic microorganisms capable of producing at least about 20% of their biomass as lipids comprising adding to a fermentation medium comprising said microorganisms a non-alcohol carbon source and a limiting nutrient source at a rate sufficient to increase the biomass density of said fermentation medium to at least about 100 g/L.
2. The process of Claim 1, wherein said process comprises a biomass density increasing stage and a production stage, wherein said biomass density increasing stage comprises adding said carbon source and said limiting nutrient source and said production stage comprises adding said carbon source with no or little additional said limiting nutrient source to induce nutrient limiting conditions which induces lipid production.
3. The process of Claim 2, wherein the amount of dissolved oxygen present in said fermentation medium during said production stage is lower than the amount of dissolved oxygen present in said fermentation medium during said biomass density increasing stage.
4. The process of Claim 3, wherein the amount of dissolved oxygen in said fermentation medium during said biomass density increasing stage is at least about 4%.
5. The process of Claim 4, wherein the amount of dissolved oxygen in said fermentation medium during said production stage is less than about 1%.
6. The process of Claim 1, wherein said non-alcohol carbon source comprises a carbohydrate.
7. The process of Claim 1, wherein said limiting nutrient source is selected from the group consisting of nitrogen sources, carbon sources, phosphate sources, vitamin sources (such as vitamin B12 sources, pantothenate sources, thiamine sources), trace metal sources (such as zinc sources, copper sources, cobalt sources, nickel sources, iron sources, manganese sources, molybdenum sources), and major metal sources (such as magnesium sources, calcium sources, sodium sources, potassium sources), silica sources and mixtures thereof.
8. The process of Claim 7, wherein said trace metal sources and major metal sources are selected from the group consisting of sulfate and chloride salts of these metals (such as MgSO4.cndot.7H2O; MnCl2.cndot.4H2O; ZnSO4.cndot.7H2O; CoCl2.cndot.6H2O;
Na2MoO4.cndot.2H2O; CuSO4.cndot.5H2O;
NiSO4.cndot.6H2O; FeSO4.cndot.7H2O; CaCl2; K2SO4; KCl; and Na2SO4) and mixtures thereof.
Na2MoO4.cndot.2H2O; CuSO4.cndot.5H2O;
NiSO4.cndot.6H2O; FeSO4.cndot.7H2O; CaCl2; K2SO4; KCl; and Na2SO4) and mixtures thereof.
9. The process of Claim 1, wherein said limiting nutrient source comprises a nitrogen source.
10. The process of Claim 9, wherein said nitrogen source comprises an inorganic ammonium salt.
11. The process of Claim 9, wherein said nitrogen source comprises ammonium hydroxide.
12. The process of Claim 10, wherein pH of the fermentation medium is controlled by said limiting nutrient source.
13. The process of Claim 1, wherein said fermentation medium is at a temperature of at least about 20°C.
14. The process of Claim 1, wherein said process produces lipids at an average rate of at least about 0.5 g/L/hr.
15. The process of Claim 14, wherein at least about 15% of said lipids are polyunsaturated lipids.
16. The process of Claim 14, wherein the total amount of omega-3 and omega-6 fatty acids is at least about 20% of said lipids.
17. The process of Claim 14, wherein at least about 25% of said lipids is docosahexaenoic acid.
18. The process of Claim 1, wherein said microorganisms are selected from the group consisting of algae, fungi (including yeast), protists, bacteria and mixtures thereof, wherein said microorganisms are capable of producing polyenoic fatty acids or other lipids which had been generally believed to require oxygen for their synthesis.
19. The process of Claim 18, wherein said microorganisms are capable of producing lipids at a fermentation medium oxygen level of about less than 3% of saturation.
20. The process of Claim 18, wherein said microorganisms are grown in a fed-batch process.
21. The process of Claim 20 further comprising maintaining an oxygen level of less than about 3% of saturation in said fermentation medium during the production phase of said fermentation process.
22. The process of Claim 1, wherein said microorganisms are microalgae and algae-like microorganisms.
23. The process of Claim 1, wherein said microorganisms are Stramenopiles.
24. The process of Claim 1, wherein said microorganisms are of the order Thraustochytriales.
25. The process of Claim 1, wherein said microorganisms are selected from the group consisting of Thraustochytrium, Schizochytrium, and mixtures thereof.
26. A process for growing eukaryotic microorganisms capable of producing at least about 20% of their biomass as lipids, said process comprising adding to a fermentation medium comprising said microorganisms a carbon source and a limiting nutrient source at a rate sufficient to increase the biomass density of said fermentation medium to at least about 100 g/L, wherein said process produces at least an average rate of about 0.5 g/L/hr of lipids, and wherein at least about 15% of the total lipids produced by said microorganisms is polyunsaturated lipids.
27. The process of Claim 26, wherein said process further comprises:
(a) a biomass density increasing stage, wherein the dissolved oxygen level in the fermentation medium is at least about 4%; and (b) a high lipid production stage, wherein the dissolved oxygen level in the fermentation medium is less than about 1%.
(a) a biomass density increasing stage, wherein the dissolved oxygen level in the fermentation medium is at least about 4%; and (b) a high lipid production stage, wherein the dissolved oxygen level in the fermentation medium is less than about 1%.
28. The process of Claim 26, wherein said carbon source is a non-alcohol carbon source.
29. The process of Claim 26, wherein said limiting nutrient source is selected from the group consisting of nitrogen sources, carbon sources, phosphate sources, vitamin sources (such as vitamin B12 sources, pantothenate sources, thiamine sources), trace metal sources (such as zinc sources, copper sources, cobalt sources, nickel sources, iron sources, manganese sources, molybdenum sources), and major metal sources (such as magnesium sources, calcium sources, sodium sources, potassium sources), and silica sources and mixtures thereof.
30. The process of Claim 29, wherein said trace metal sources and major metal sources are selected from the group consisting of sulfate and chloride salts of these metals (such as MgSO4.cndot.7H2O; MnCl2.cndot.4H2O; ZnSO4.cndot.7H2O; CoCl2.cndot.6H2O;
Na2MoO4.cndot.2H2O; CuSO4.cndot.SH2O;
NiSO4.cndot.6H2O; FeSO4.cndot.7H2O; CaC12; K2SO4; KCl; and Na2SO4) and mixtures thereof.
Na2MoO4.cndot.2H2O; CuSO4.cndot.SH2O;
NiSO4.cndot.6H2O; FeSO4.cndot.7H2O; CaC12; K2SO4; KCl; and Na2SO4) and mixtures thereof.
31. The process of Claim 26, wherein said limiting nutrient source comprises a nitrogen source.
32. The process of Claim 31, wherein said nitrogen source comprises an inorganic ammonium salt.
33. The process of Claim 26, wherein said microorganisms are selected from the group consisting of algae, fungi (including yeast), protists, bacteria and mixtures thereof, wherein said microorganisms are capable of producing polyenoic fatty acids or other lipids which had been generally believed to requireoxygen for their synthesis.
34. The process of Claim 26, wherein said microorganisms are Stramenopiles.
35. The process of Claim 26, wherein said microorganisms are microalgae.
36. The process of Claim 26, wherein said microorganisms are of the order Thraustochytriales.
37. The process of Claim 26, wherein said microorganisms are selected from the group consisting of Thnaustochytrium, Schizochytrium, and mixtures thereof.
38. The process of Claim 26, wherein the total amount of omega-3 and omega-6 fatty acids is at least about 20 % of said lipids.
39. The process of Claim 26, wherein at least about 25 % of said lipids is docosahexaenoic acid.
40. A process for growing eukaryotic microorganisms comprising adding to a fermentation medium comprising said microorganisms a carbon source and a limiting nutrient source at a rate sufficient to increase the biomass density of said fermentation medium to at least about 100 g/L, wherein said microorganisms are selected from the group consisting of algae, fungi (including yeast), protists, bacteria and mixtures thereof, and wherein said microorganisms are capable of producing at least about 20% of their biomass as lipids comprising some polyenoic fatty acids.
41. The process of Claim 40, wherein said process produces microbial lipids at an average rate of least about 0.5 g/L/hr.
42. The process of Claim 41, wherein greater than about 15% of said lipids are polyunsaturated lipids.
43. The process of Claim 41, wherein the total amount of omega-3 and omega-6 polyunsaturated fatty acids are at least about 20% of said lipids.
44. The process of Claim 41, wherein at least about 25% of said lipids is docosahexaenoic acid.
45. The process of Claim 40, wherein said process produces on average at least about 0.2 g/L/hr of docosahexaenoic acid.
46. The process of Claim 40, wherein said carbon source is not an alcohol.
47. The process of Claim 46, wherein said carbon source is a carbohydrate.
48. The process of Claim 40, wherein said limiting nutrient source is selected from the group consisting of nitrogen sources, carbon sources, phosphate sources, vitamin sources (such as vitamin B12 sources, pantothenate sources, thiamine sources), trace metal sources (such as zinc sources, copper sources, cobalt sources, nickel sources, iron sources, manganese sources, molybdenum sources), and major metal sources (such as magnesium sources, calcium sources, sodium sources, potassium sources), silica sources, and mixtures thereof.
49. The process of Claim 48, wherein said trace metal sources and major metal sources are selected from the group consisting of sulfate and chloride salts of these metals (such as MgSO4.cndot.7H2O; MnCl2.cndot.4H2O; ZnSO4.cndot.7H2O; CoCl2.cndot.6H2O;
Na2MoO4.cndot.2H2O; CuSO4.cndot.5H2O;
NiSO4.cndot.6H2O; FeSO4.cndot.7H2O; CaCl2; K2SO4; KCl; and Na2SO4) and mixtures thereof.
Na2MoO4.cndot.2H2O; CuSO4.cndot.5H2O;
NiSO4.cndot.6H2O; FeSO4.cndot.7H2O; CaCl2; K2SO4; KCl; and Na2SO4) and mixtures thereof.
50. The process of Claim 40, wherein said limiting nutrient sources comprises a nitrogen source.
51. The process of Claim 50, wherein said nitrogen source comprises an inorganic ammonium salt.
52. The process of Claim 40, wherein said nitrogen source comprises ammonium hydroxide.
53. The process of Claim 40, wherein said microorganisms are selected from the group consisting of algae, fungi (including yeast), protists, bacteria and mixtures thereof, wherein said microorganisms are capable of producing polyenoic fatty acids or other lipids which had been generally believed to require oxygen for their synthesis.
54. The process of Claim 40, wherein said microorganisms are microalgae and algae-like microorganisms.
55. The process of Claim 40, wherein said microorganisms are Stramenopiles.
56. A process for producing eukaryotic microbial lipids comprising:
(a) growing eukaryotic microorganisms in a fermentation medium to increase the biomass density of said fermentation medium to at least about 100 g/L;
(b) providing fermentation conditions sufficient to allow said microorganisms to produce said lipids; and (c) recovering said lipids,wherein greater than about 15% of said lipids are polyunsaturated lipids.
(a) growing eukaryotic microorganisms in a fermentation medium to increase the biomass density of said fermentation medium to at least about 100 g/L;
(b) providing fermentation conditions sufficient to allow said microorganisms to produce said lipids; and (c) recovering said lipids,wherein greater than about 15% of said lipids are polyunsaturated lipids.
57. The process of Claim 56, wherein a dissolved oxygen level in said fermentation medium during said microorganism growing step is higher than the dissolved oxygen level in the fermentation medium during said lipid producing step.
58. The process of Claim 56, wherein the dissolved oxygen level in said fermentation medium during the microorganism growing step is at least about 4%.
59. The process of Claim 56, wherein the dissolved oxygen level in said fermentation medium during the lipid producing step is less than about 1%.
60. The process of Claim 56, wherein said microorganisms are selected from the group consisting of algae, fungi (including yeast), protists, bacteria and mixtures thereof, wherein said microorganisms are capable of producing polyenoic fatty acids or other lipids which had been generally believed to require oxygen for their synthesis.
61. The process of Claim 56, wherein said microorganisms are Stramenopiles.
62. The process of Claim 56, wherein said microorganisms are microalgae and algae-like microorganisms.
63. The process of Claim 56, wherein said microorganisms are of the order Thraustochytriales.
64. The process of Claim 56, wherein said microorganisms are selected from the group consisting of Thraustochytrium, Schizochytrium, and mixtures thereof.
65. The process of Claim 56, wherein the total amount of omega-3 and omega-6 fatty acids is at least about 20% of said microbial lipids.
66. The process of Claim 56, wherein at least about 25% of said microbial lipids is docosahexaenoic acid.
67. The process of Claim 56, wherein said process produces docosahexaenoic acid at an average rate of at least about 0.2 g/L/hr.
68. The process of Claim 56, wherein said lipid recovery step comprises:
(d) removing water from said fermentation medium to provide dry microorganisms; and (e) isolating said lipids from said dry microorganisms.
(d) removing water from said fermentation medium to provide dry microorganisms; and (e) isolating said lipids from said dry microorganisms.
69. The process of Claim 68, wherein said water removal step comprises contacting said fermentation medium directly on a drum-dryer without prior centrifugation.
70. The process of Claim 56, wherein said lipid recovery step comprises:
(d) treating the fermentation broth to permeabilize, lyse or rupture the microbial cells; and (e) recovering the lipids from the fermentation broth by gravity separation, with or without the aid of anagent to aid in breaking the lipid/water emulsion.
(d) treating the fermentation broth to permeabilize, lyse or rupture the microbial cells; and (e) recovering the lipids from the fermentation broth by gravity separation, with or without the aid of anagent to aid in breaking the lipid/water emulsion.
71. The process of Claim 70, wherein said microbial cells are treated in step (d) in a fermentor or a similar vessel.
72. The process of Claim 70, wherein said gravity separation comprises centrifugation.
73. The process of Claim 56, wherein said microorganism growing step comprises adding a carbon source and a limiting nutrient source.
74. The process of Claim 73, wherein said carbon source is non-alcohol.
75. The process of Claim 74, wherein said carbon source comprises a carbohydrate.
76. The process of Claim 73, wherein said limiting nutrient source is selected from the group consisting of nitrogen sources, carbon sources, phosphate sources, vitamin sources (such as vitamin B12 sources, pantothenate sources, thiamine sources), trace metal sources (such as zinc sources, copper sources, cobalt sources, nickel sources, iron sources, manganese sources, molybdenum sources), and major metal sources (such as magnesium sources, calcium sources, sodium sources, potassium sources), silica sources, and mixtures thereof.
77. The process of Claim 76, wherein said trace metal sources and major metal sources are selected from the group consisting of sulfate and chloride salts of these metals (such as MgSO4.cndot.7H2O; MnCl2.cndot.4H2O; ZnSO4.cndot.7H2O; CoCl2.cndot.6H2O ;
Na2MoO4.cndot.2H2O; CuSO4.cndot.5H2O;
NiSO4.cndot.6H2O ; FeSO4.cndot.7H2O ; CaCl2; K2SO4; KCl; and Na2SO4) and mixtures thereof.
Na2MoO4.cndot.2H2O; CuSO4.cndot.5H2O;
NiSO4.cndot.6H2O ; FeSO4.cndot.7H2O ; CaCl2; K2SO4; KCl; and Na2SO4) and mixtures thereof.
78. The process of Claim 73, wherein said limiting nutrient source comprises a nitrogen source.
79. The process of Claim 78, wherein said nitrogen source comprises an inorganic ammonium salt.
80. The process of Claim 78, wherein said nitrogen source comprises ammonium hydroxide.
81. A process for producing eukaryotic microbial lipids comprising:
(a) adding a non-alcohol carbon source and a limiting nutrient source to a fermentation medium comprising said microorganisms;
(b) providing conditions sufficient for said microorganisms to produce said microbial lipids; and (c) recovering said microbial lipids, wherein at least about 15% of said microbial lipids are polyunsaturated lipids.
(a) adding a non-alcohol carbon source and a limiting nutrient source to a fermentation medium comprising said microorganisms;
(b) providing conditions sufficient for said microorganisms to produce said microbial lipids; and (c) recovering said microbial lipids, wherein at least about 15% of said microbial lipids are polyunsaturated lipids.
82. The process of Claim 81, wherein the dissolved oxygen level in said fermentation medium during a biomass density increasing step is higher than the dissolved oxygen level in said fermentation medium during a lipid producing step.
83. The process of Claim 81, wherein the dissolved oxygen level in said fermentation medium during a biomass density increasing step is at least about 4%.
84. The process of Claim 81, wherein the dissolved oxygen level in said fermentation medium during a lipid producing step is about 1% or less.
85. The process of Claim 81, wherein said microorganisms are selected from the group consisting of algae, fungi (including yeast), protists, bacteria and mixtures thereof.
86. The process of Claim 81, wherein said microorganisms are selected from the group consisting of algae, fungi (including yeast), protists, bacteria and mixtures thereof, wherein said microorganisms are capable of producing polyenoic fatty acids or other lipids which had been generally believed to require oxygen for their synthesis.
87. The process of Claim 81, wherein said microorganisms are Stramenopiles.
88. The process of Claim 81, wherein said microorganisms are microalgae and algae-like microorganisms.
89. The process of Claim 81, wherein said microorganisms are of the order Thraustochytriales.
90. The process of Claim 81, wherein said microorganisms are selected from the group consisting of Thraustochytrium, Schizochytrium, and mixtures thereof.
91. The process of Claim 81, wherein said process produces said microbial lipids at an average rate of at least about 0.5 g/L/hr.
92. The process of Claim 91, wherein the total amount of omega-3 and omega-6 fatty acids is at least about 20% of said microbial lipids.
93. The process of Claim 81, wherein at least about 25% of said lipids is docosahexaenoic acid.
94. The process of Claim 81, wherein said process produces docosahexaenoic acid at an average rate of at least about 0.2 g/L/hr.
95. The process of Claim 58, wherein said lipid recovery step comprises:
(d) removing water from said fermentation medium to provide dry microorganisms; and (e) isolating said lipids from said dry microorganisms.
(d) removing water from said fermentation medium to provide dry microorganisms; and (e) isolating said lipids from said dry microorganisms.
96. The process of Claim 95, wherein said water removal step comprises evaporating water from said fermentation medium without prior centrifugation.
97. The process of Claim 81, wherein said lipid recovery step comprises:
(d) treating the fermentation broth to permeabilize, lyse or rupture the microbial cells; and (e) recovering the lipids from the fermentation broth by gravity separation, with or without the aid of an agent to aid in breaking the lipid/water emulsion.
(d) treating the fermentation broth to permeabilize, lyse or rupture the microbial cells; and (e) recovering the lipids from the fermentation broth by gravity separation, with or without the aid of an agent to aid in breaking the lipid/water emulsion.
98. The process of Claim 97, wherein said microbial cells are treated in step (d) in a fermentor or a similar vessel.
99. The process of Claim 97, wherein said gravity separation comprises centrifugation.
100. The process of Claim 81, wherein said non-alcohol carbon source comprises a carbohydrate.
101. The process of Claim 81, wherein said limiting nutrient source is selected from the group consisting of nitrogen sources, carbon sources, phosphate sources, vitamin sources (such as vitamin B12 sources, pantothenate sources, thiamine sources), trace metal sources (such as zinc sources, copper sources, cobalt sources, nickel sources, iron sources, manganese sources, molybdenum sources), and major metal sources (such as magnesium sources, calcium sources, sodium sources, potassium sources), silica sources, and mixtures thereof.
102. The process of Claim 101, wherein said trace metal sources and major metal sources are selected from the group consisting of sulfate and chloride salts of these metals (such as MgSO4.cndot.7H2O; MnCl2.cndot.4H2O; ZnSO4.cndot.7H2O; CoCl2.cndot.6H2O;
Na2MoO4.cndot.2H2O; CuSO4.cndot.5H2O ;
NiSO4.cndot.6H2O; FeSO4.cndot.7H2O; CaCl2; K2SO4; KCl; and Na2SO4) and mixtures thereof.
Na2MoO4.cndot.2H2O; CuSO4.cndot.5H2O ;
NiSO4.cndot.6H2O; FeSO4.cndot.7H2O; CaCl2; K2SO4; KCl; and Na2SO4) and mixtures thereof.
103. The process of Claim 81, wherein said limiting nutrient source comprises a nitrogen source.
104. The process of Claim 103, wherein said nitrogen source comprises an inorganic ammonium salt.
105. The process of Claim 103, wherein said nitrogen source comprises ammonium hydroxide.
106. A process for producing eukaryotic microbial lipids comprising:
(a) adding a carbon source and a limiting nutrient source to a fermentation medium comprising eukaryotic microorganisms and providing conditions sufficient to maintain a dissolved oxygen level of at least about 4% in said fermentation medium to produce a biomass density of at least about 100 g/L;
(b) providing conditions sufficient to maintain a dissolved oxygen level of about 1% or less in said fermentation medium and providing conditions sufficient to allow said microorganisms to produce said lipids; and (c) recovering said microbial lipids, wherein at least about 15% of said microbial lipids are polyunsaturated lipids.
(a) adding a carbon source and a limiting nutrient source to a fermentation medium comprising eukaryotic microorganisms and providing conditions sufficient to maintain a dissolved oxygen level of at least about 4% in said fermentation medium to produce a biomass density of at least about 100 g/L;
(b) providing conditions sufficient to maintain a dissolved oxygen level of about 1% or less in said fermentation medium and providing conditions sufficient to allow said microorganisms to produce said lipids; and (c) recovering said microbial lipids, wherein at least about 15% of said microbial lipids are polyunsaturated lipids.
107. The process of Claim 106, wherein said microorganisms are selected from the group consisting of algae, fungi (including yeast), protists, bacteria and mixtures thereof.
108. The process of Claim 106, wherein said microorganisms are selected from the group consisting of algae, fungi (including yeast), protists, bacteria and mixtures thereof, wherein said microorganisms are capable of producing polyenoic fatty acids or other lipids which had been generally believed to require oxygen for their synthesis.
109. The process of Claim 106, wherein said microorganisms are Stramenopiles.
110. The process of Claim 106, wherein said microorganisms are microalgae and algae-like microorganisms.
111. The process of Claim 106, wherein said microorganisms are of the order Thraustochytriales.
112. The process of Claim 106, wherein said microorganisms are selected from the group consisting of Thraustochytrium, Schizochytrium, and mixtures thereof.
113. The process of Claim 106, wherein at least about 20% of said microbial lipids is omega-3 and/or omega-6 fatty acids.
114. The process of Claim 106, wherein at least about 25% of said microbial lipids is docosahexaenoic acid.
115. The process of Claim 106, wherein said process produces docosahexaenoic acid at an average rate of at least about 0.2 g/L/hr.
116. The process of Claim 106, wherein the lipid recovery step comprises:
(d) removing water from said fermentation medium to provide dry microorganisms; and (e) isolating said lipids from said dry microorganisms.
(d) removing water from said fermentation medium to provide dry microorganisms; and (e) isolating said lipids from said dry microorganisms.
117. The process of Claim 116, wherein the water removal step comprises evaporating the water directly from said fermentation medium without prior centrifugation.
118. The process of Claim 106, wherein the lipid recovery step comprises:
(d) treating the fermentation broth to permeabilize, lyse or rupture the microbial cells; and (e) recovering the lipids from the fermentation broth by gravity separation, with or without the aid of a agent to aid in breaking the lipid/water emulsion.
(d) treating the fermentation broth to permeabilize, lyse or rupture the microbial cells; and (e) recovering the lipids from the fermentation broth by gravity separation, with or without the aid of a agent to aid in breaking the lipid/water emulsion.
119. The process of Claim 106, wherein said microbial cells are treated in step (d) in a fermentor or a similar vessel.
120. The process of Claim 118, wherein said gravity separation comprises centrifugation.
121. A method for enriching the polyenoic fatty acid content of a microorganism comprising fermenting said microorganism in a growth medium having a level of dissolved oxygen of less than 10%.
122. The process of Claim 121, wherein said level of dissolved oxygen is less than 5 percent.
123. The process of Claim 121, wherein said level of dissolved oxygen is less than 1 percent.
124. The process of Claim 121, wherein during an initial growth phase, the level of dissolved oxygen is greater than about 10 percent and during a subsequent production phase the level of dissolved oxygen is less than 10 percent.
125. The process of Claim 121, wherein said microorganism contains polyketide synthase genes.
126. A heterotrophic process for producing products and microorganisms comprising:
a. culturing said microorganisms in a growth medium wherein said microorganisms contain polyketide synthase genes; and b. maintaining the level of dissolved oxygen at less than about 10 percent.
a. culturing said microorganisms in a growth medium wherein said microorganisms contain polyketide synthase genes; and b. maintaining the level of dissolved oxygen at less than about 10 percent.
127. The process of Claim 126, wherein said level of dissolved oxygen is less than about 5 percent.
128. The process of Claim 126, wherein said level of dissolved oxygen is less than about 1 percent.
129. The process of Claim 126, wherein said polyketide synthase genes occur naturally in said microorganisms.
130. The process of Claim 126, wherein said polyketide synthase genes are genetically introduced into the said microorganisms.
131. The process of Claim 126, wherein during an initial growth phase, the level of dissolved oxygen is greater than about 10 percent and during a subsequent production phase the level of dissolved oxygen is less than about 10 percent.
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CA2786722A CA2786722A1 (en) | 2000-01-28 | 2001-01-26 | Enhanced production of lipids containing polyenoic fatty acids by high density cultures of eukaryotic microbes in fermentors |
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US5340742A (en) * | 1988-09-07 | 1994-08-23 | Omegatech Inc. | Process for growing thraustochytrium and schizochytrium using non-chloride salts to produce a microfloral biomass having omega-3-highly unsaturated fatty acids |
US20060094089A1 (en) * | 1988-09-07 | 2006-05-04 | Martek Biosciences Corporation | Process for the heterotrophic production of microbial products with high concentrations of omega-3 highly unsaturated fatty acids |
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2002
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2003
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2005
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2006
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2007
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