CA2101274C - Microbial oil mixtures and uses thereof - Google Patents
Microbial oil mixtures and uses thereofInfo
- Publication number
- CA2101274C CA2101274C CA002101274A CA2101274A CA2101274C CA 2101274 C CA2101274 C CA 2101274C CA 002101274 A CA002101274 A CA 002101274A CA 2101274 A CA2101274 A CA 2101274A CA 2101274 C CA2101274 C CA 2101274C
- Authority
- CA
- Canada
- Prior art keywords
- dha
- ara
- oil
- parts
- composition
- 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.)
- Expired - Lifetime
Links
- 239000000203 mixture Substances 0.000 title claims abstract description 149
- 230000000813 microbial effect Effects 0.000 title claims abstract description 57
- 239000003921 oil Substances 0.000 claims abstract description 193
- 235000013350 formula milk Nutrition 0.000 claims abstract description 74
- 238000000034 method Methods 0.000 claims abstract description 29
- 239000013589 supplement Substances 0.000 claims abstract description 15
- YZXBAPSDXZZRGB-DOFZRALJSA-N arachidonic acid Chemical compound CCCCC\C=C/C\C=C/C\C=C/C\C=C/CCCC(O)=O YZXBAPSDXZZRGB-DOFZRALJSA-N 0.000 claims description 251
- 235000019198 oils Nutrition 0.000 claims description 191
- 235000021342 arachidonic acid Nutrition 0.000 claims description 125
- 229940114079 arachidonic acid Drugs 0.000 claims description 125
- 235000020256 human milk Nutrition 0.000 claims description 37
- 235000020664 gamma-linolenic acid Nutrition 0.000 claims description 24
- VZCCETWTMQHEPK-QNEBEIHSSA-N gamma-linolenic acid Chemical compound CCCCC\C=C/C\C=C/C\C=C/CCCCC(O)=O VZCCETWTMQHEPK-QNEBEIHSSA-N 0.000 claims description 23
- 235000021323 fish oil Nutrition 0.000 claims description 19
- VZCCETWTMQHEPK-UHFFFAOYSA-N gamma-Linolensaeure Natural products CCCCCC=CCC=CCC=CCCCCC(O)=O VZCCETWTMQHEPK-UHFFFAOYSA-N 0.000 claims description 15
- 229960002733 gamolenic acid Drugs 0.000 claims description 15
- 235000015872 dietary supplement Nutrition 0.000 claims description 11
- 238000002156 mixing Methods 0.000 claims description 10
- 239000002253 acid Substances 0.000 claims description 9
- 150000003626 triacylglycerols Chemical class 0.000 claims description 9
- 241000197220 Pythium insidiosum Species 0.000 claims description 8
- 241000907999 Mortierella alpina Species 0.000 claims description 7
- 238000004519 manufacturing process Methods 0.000 claims description 7
- 241000235395 Mucor Species 0.000 claims description 4
- 241000245063 Primula Species 0.000 claims description 4
- 235000016311 Primula vulgaris Nutrition 0.000 claims description 4
- 239000010473 blackcurrant seed oil Substances 0.000 claims description 4
- 230000000474 nursing effect Effects 0.000 claims description 4
- 241000894007 species Species 0.000 claims description 4
- 230000001502 supplementing effect Effects 0.000 claims description 4
- 241000306282 Umbelopsis isabellina Species 0.000 claims description 3
- 238000006243 chemical reaction Methods 0.000 claims description 3
- UFTFJSFQGQCHQW-UHFFFAOYSA-N triformin Chemical compound O=COCC(OC=O)COC=O UFTFJSFQGQCHQW-UHFFFAOYSA-N 0.000 claims 11
- DTOSIQBPPRVQHS-PDBXOOCHSA-N alpha-linolenic acid Chemical compound CC\C=C/C\C=C/C\C=C/CCCCCCCC(O)=O DTOSIQBPPRVQHS-PDBXOOCHSA-N 0.000 claims 9
- 235000020661 alpha-linolenic acid Nutrition 0.000 claims 9
- 229960004488 linolenic acid Drugs 0.000 claims 9
- KQQKGWQCNNTQJW-UHFFFAOYSA-N linolenic acid Natural products CC=CCCC=CCC=CCCCCCCCC(O)=O KQQKGWQCNNTQJW-UHFFFAOYSA-N 0.000 claims 9
- 241001072256 Boraginaceae Species 0.000 claims 2
- 235000007689 Borago officinalis Nutrition 0.000 claims 2
- 241000199913 Crypthecodinium Species 0.000 claims 2
- 239000010474 borage seed oil Substances 0.000 claims 2
- 235000021476 total parenteral nutrition Nutrition 0.000 claims 1
- 235000020978 long-chain polyunsaturated fatty acids Nutrition 0.000 abstract description 8
- 235000020777 polyunsaturated fatty acids Nutrition 0.000 description 27
- 210000004027 cell Anatomy 0.000 description 18
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 18
- JAZBEHYOTPTENJ-JLNKQSITSA-N all-cis-5,8,11,14,17-icosapentaenoic acid Chemical compound CC\C=C/C\C=C/C\C=C/C\C=C/C\C=C/CCCC(O)=O JAZBEHYOTPTENJ-JLNKQSITSA-N 0.000 description 16
- 235000020673 eicosapentaenoic acid Nutrition 0.000 description 16
- 229960005135 eicosapentaenoic acid Drugs 0.000 description 16
- JAZBEHYOTPTENJ-UHFFFAOYSA-N eicosapentaenoic acid Natural products CCC=CCC=CCC=CCC=CCC=CCCCC(O)=O JAZBEHYOTPTENJ-UHFFFAOYSA-N 0.000 description 16
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 15
- 239000008103 glucose Substances 0.000 description 14
- 239000002028 Biomass Substances 0.000 description 12
- 235000015112 vegetable and seed oil Nutrition 0.000 description 12
- 239000008158 vegetable oil Substances 0.000 description 12
- 235000014113 dietary fatty acids Nutrition 0.000 description 11
- 239000003925 fat Substances 0.000 description 11
- 235000019197 fats Nutrition 0.000 description 11
- 229930195729 fatty acid Natural products 0.000 description 11
- 239000000194 fatty acid Substances 0.000 description 11
- 150000004665 fatty acids Chemical class 0.000 description 11
- 239000000654 additive Substances 0.000 description 9
- 210000004251 human milk Anatomy 0.000 description 9
- 238000013019 agitation Methods 0.000 description 8
- 238000002360 preparation method Methods 0.000 description 8
- 241000199912 Crypthecodinium cohnii Species 0.000 description 7
- 239000002609 medium Substances 0.000 description 7
- 241000273930 Brevoortia tyrannus Species 0.000 description 6
- 230000000996 additive effect Effects 0.000 description 6
- 150000002632 lipids Chemical class 0.000 description 6
- 239000006188 syrup Substances 0.000 description 6
- 235000020357 syrup Nutrition 0.000 description 6
- 229940013317 fish oils Drugs 0.000 description 5
- 239000008188 pellet Substances 0.000 description 5
- 241001474374 Blennius Species 0.000 description 4
- OYHQOLUKZRVURQ-HZJYTTRNSA-N Linoleic acid Chemical compound CCCCC\C=C/C\C=C/CCCCCCCC(O)=O OYHQOLUKZRVURQ-HZJYTTRNSA-N 0.000 description 4
- 241000206745 Nitzschia alba Species 0.000 description 4
- BPQQTUXANYXVAA-UHFFFAOYSA-N Orthosilicate Chemical compound [O-][Si]([O-])([O-])[O-] BPQQTUXANYXVAA-UHFFFAOYSA-N 0.000 description 4
- 238000005273 aeration Methods 0.000 description 4
- 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 description 4
- 235000005911 diet Nutrition 0.000 description 4
- 235000020778 linoleic acid Nutrition 0.000 description 4
- OYHQOLUKZRVURQ-IXWMQOLASA-N linoleic acid Natural products CCCCC\C=C/C\C=C\CCCCCCCC(O)=O OYHQOLUKZRVURQ-IXWMQOLASA-N 0.000 description 4
- 102000002322 Egg Proteins Human genes 0.000 description 3
- 108010000912 Egg Proteins Proteins 0.000 description 3
- 235000008452 baby food Nutrition 0.000 description 3
- 239000010779 crude oil Substances 0.000 description 3
- 235000013345 egg yolk Nutrition 0.000 description 3
- 210000002969 egg yolk Anatomy 0.000 description 3
- 229910052751 metal Inorganic materials 0.000 description 3
- 239000002184 metal Substances 0.000 description 3
- 150000002739 metals Chemical class 0.000 description 3
- 235000013336 milk Nutrition 0.000 description 3
- 239000008267 milk Substances 0.000 description 3
- 210000004080 milk Anatomy 0.000 description 3
- 235000020665 omega-6 fatty acid Nutrition 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 239000011550 stock solution Substances 0.000 description 3
- 230000009469 supplementation Effects 0.000 description 3
- 239000008399 tap water Substances 0.000 description 3
- 235000020679 tap water Nutrition 0.000 description 3
- 235000003563 vegetarian diet Nutrition 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 241000282412 Homo Species 0.000 description 2
- 241000233639 Pythium Species 0.000 description 2
- 240000001890 Ribes hudsonianum Species 0.000 description 2
- 235000016954 Ribes hudsonianum Nutrition 0.000 description 2
- 235000001466 Ribes nigrum Nutrition 0.000 description 2
- 235000021324 borage oil Nutrition 0.000 description 2
- 210000000481 breast Anatomy 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 239000002537 cosmetic Substances 0.000 description 2
- 230000000994 depressogenic effect Effects 0.000 description 2
- 230000037213 diet Effects 0.000 description 2
- 230000000378 dietary effect Effects 0.000 description 2
- 235000020669 docosahexaenoic acid Nutrition 0.000 description 2
- 229940090949 docosahexaenoic acid Drugs 0.000 description 2
- 235000006486 human diet Nutrition 0.000 description 2
- 239000002054 inoculum Substances 0.000 description 2
- 235000020667 long-chain omega-3 fatty acid Nutrition 0.000 description 2
- 230000002503 metabolic effect Effects 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 239000011785 micronutrient Substances 0.000 description 2
- 235000013369 micronutrients Nutrition 0.000 description 2
- 235000016709 nutrition Nutrition 0.000 description 2
- 235000020660 omega-3 fatty acid Nutrition 0.000 description 2
- 210000000056 organ Anatomy 0.000 description 2
- 150000003904 phospholipids Chemical class 0.000 description 2
- 239000002243 precursor Substances 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Chemical compound [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 2
- 235000013343 vitamin Nutrition 0.000 description 2
- 239000011782 vitamin Substances 0.000 description 2
- 229940088594 vitamin Drugs 0.000 description 2
- 229930003231 vitamin Natural products 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- KEZYHIPQRGTUDU-UHFFFAOYSA-N 2-[dithiocarboxy(methyl)amino]acetic acid Chemical compound SC(=S)N(C)CC(O)=O KEZYHIPQRGTUDU-UHFFFAOYSA-N 0.000 description 1
- 241000251468 Actinopterygii Species 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241000235575 Mortierella Species 0.000 description 1
- 229910003424 Na2SeO3 Inorganic materials 0.000 description 1
- 241000180701 Nitzschia <flatworm> Species 0.000 description 1
- KUGRPPRAQNPSQD-UHFFFAOYSA-N OOOOO Chemical compound OOOOO KUGRPPRAQNPSQD-UHFFFAOYSA-N 0.000 description 1
- MOMWFXLCFJOAFX-UHFFFAOYSA-N OOOOOOOO Chemical compound OOOOOOOO MOMWFXLCFJOAFX-UHFFFAOYSA-N 0.000 description 1
- 244000061456 Solanum tuberosum Species 0.000 description 1
- 235000002595 Solanum tuberosum Nutrition 0.000 description 1
- JZRWCGZRTZMZEH-UHFFFAOYSA-N Thiamine Natural products CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N JZRWCGZRTZMZEH-UHFFFAOYSA-N 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 210000001557 animal structure Anatomy 0.000 description 1
- 239000002518 antifoaming agent Substances 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 235000015278 beef Nutrition 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 239000008121 dextrose Substances 0.000 description 1
- 238000004821 distillation Methods 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- -1 fish oils Substances 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 239000007903 gelatin capsule Substances 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 235000010598 long-chain omega-6 fatty acid Nutrition 0.000 description 1
- 239000004570 mortar (masonry) Substances 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 230000005305 organ development Effects 0.000 description 1
- 238000007911 parenteral administration Methods 0.000 description 1
- 235000015277 pork Nutrition 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 239000013535 sea water Substances 0.000 description 1
- 235000010344 sodium nitrate Nutrition 0.000 description 1
- 239000011781 sodium selenite Substances 0.000 description 1
- 235000015921 sodium selenite Nutrition 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 235000019157 thiamine Nutrition 0.000 description 1
- KYMBYSLLVAOCFI-UHFFFAOYSA-N thiamine Chemical compound CC1=C(CCO)SCN1CC1=CN=C(C)N=C1N KYMBYSLLVAOCFI-UHFFFAOYSA-N 0.000 description 1
- 229960003495 thiamine Drugs 0.000 description 1
- 239000011721 thiamine Substances 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 230000008467 tissue growth Effects 0.000 description 1
- 230000002792 vascular Effects 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Classifications
-
- 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
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/21—Esters, e.g. nitroglycerine, selenocyanates
- A61K31/215—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids
- A61K31/22—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids of acyclic acids, e.g. pravastatin
- A61K31/23—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids of acyclic acids, e.g. pravastatin of acids having a carboxyl group bound to a chain of seven or more carbon atoms
-
- 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
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/115—Fatty acids or derivatives thereof; Fats or oils
- A23L33/12—Fatty acids or derivatives thereof
-
- 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
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/40—Complete food formulations for specific consumer groups or specific purposes, e.g. infant formula
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
- A61K31/19—Carboxylic acids, e.g. valproic acid
- A61K31/20—Carboxylic acids, e.g. valproic acid having a carboxyl group bound to a chain of seven or more carbon atoms, e.g. stearic, palmitic, arachidic acids
-
- 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
-
- 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
Abstract
The present invention relates to compositions including blends of microbial oils, methods of using such compositions, particularly as supplements for infant formula, and methods of increasing the amount of long chain polyunsaturated fatty acids in infant formula.
Description
W~92/12711 PCT/US92/~522 21012~
MICROBIAL OIL MIXTURES AND USES THEREOF
This invention relates to blends or mixtures of polyunsaturated fatty acid-contA;ni~g microbial oils and to uses thereof. In a specific preferred embodiment, this invention concerns the use of such oils as an additive or supplement for human diets, for example, as an additive to infant formula.
It long has been known that long chain polyunsaturated fatty acids (PUFAs) are essential to the human diet, particularly during periods of rapid tissue growth. Sanders et al, Am. J. Clin. Nutr., 31:805-813 (1978). Certain of these long chain acids, such as arachidonic acid (ARA), cannot be synthesized de novo in humans. Only by metabolizing linoleic acid (LOA), which is converted to gamma linolenic acid (GLA), and then to ARA can the human body produce AR~.
LOA, in turn, is an essential acid which can only be obtained from dietary sources. Additionally, the presence of eicosapentaenoic acid (EPA) in the diet inhibits the metabolic conversion of LOA to ARA.
Carlson, et al., INFORM, 1:306 (1990). ARA and docosahexaneoic acid (DHA) are critical elements of muscle, organ and vascular tissues.
Infancy is the most significant period of rapid growth in a human's life. An infant can increase its body weight by three times or more during its first year of life. Accordingly, it is critical that the infant receive adequate amounts of PUFAs to insure "~,.
proper structural and organ development. Human breast milk contains high levels of PUFAs in which the ratio of ARA to EPA is typically about 20:1. However, many women choose not to breast feed their infants for either part or all of the first year of the infant's life.
A~ recognized by Cl~n~ln;n et al., U.S. Patent 4,670,285, available infant formulas are deficient in long chain (C20 and C22) PUFAs. Cl~n~;n;n et al.
disclose an infant formula prepared from a blend of vegetable oil and egg yolk lipid and/or fish oil which can provide a total fat composition comparable to that of human breast milk. A preferable composition comprise~ from 75 to 95 parts by weight egg yolk and 5 to 25 part~ vegetable oil. This composition iB the entire lipid content of the infant formula and it is not economical to prepare. Additionally, the infant formula disclosed by Cl~n~; n; n et al. results in an EPA
level which i~ 16 times higher than the level of EPA in human breast milk and an ARA level which i8 only one quarter that of brea~t milk.
DE 3603000A1 (Milupa) discloses a computer profile of a highly polyunsaturated acid fat mixture and discusses the use of such a mixture to produce infant formulas. Sources of the fatty acids are listed as certain types of macroalgae (i.e. seaweed), fish oil, organ fats from beef and pork, and highly refined egg yolk oil. In addition to DHA from fish oil, a potential ~ource of DHA and ARA i9 said to be macroalgae, but only of the seaweed types. There is no suggestion to use microbe~ of any type, much less microbial oil.
Methods of producing microbial oils are disclosed in the following references WO/91/14427, published 2 7 ~
~_ 3 October 3, 1991 discloses the production of eicosapentaneoic acid-cont~;n;ng single cell oils (EPASCO). WO 91/11918, published August 22, 1991 discloses the production of docosahexaneoic acid-containing single cell oil (DHASCO). C~nA~;an PatentApplication No. 2,101,273 filed January 22, 1992 (WO
92/13086 published August 6, 1992) relates to the production of arachidonic acid-containing single cell oil (ARASCO). EP 322,227 also discloses a microbial oil production system. None of these references teach the use of blends containing unmodified microbial oils a~ a dietary supplement, or the use of a blend of microbial oils as an additive to exi~ting infant formula to provide that formula with a long chain PUFA
composition similar to breast milk.
Accordingly, it is an object of the present invention to provide a PUFA-enriched additive, the composition of which when added to commercial infant formula will provide desired long chain PUFAs in amounts comparable to the amounts of those PUFAs found in human breast milk.
It is an additional object of the present invention to provide an economical method of producing the above-described composition.
These, and other, objects are satisfied by the present invention as described herein.
Sllmm~ry of the Tnv~nt;on Thi~ invention relates to the use of microbial oils which contain long chain polyunsaturated fatty acids. Additionally, in various embodiments, fish oil and/or vegetable oils can be blended with such microbial oils to form desired compositions. The ~,~
7 ~ ~
_ 4 compositions can be used a~ dietary supplements, particularly as additives for infant formula, as well as for pharmaceutical and cosmetic applications.
The invention also relates to economically viable 5 processes for altering the long chain polyunsaturated fatty acid composition of infant formula and/or baby food. Preferably, the altered composition resembles that of human breast milk.
10Detailed Description of the Preferred ~mhotl;mF~nt of t h ~? TnV~nt;~n ~ roadly stated, the present invention concerns blends, or mixtures, cont~;n;ng unmodified microbial oils. As used herein, "unmodified" mean~ not 15 chemically or covalently altered. It will be understood that throughout this specification references to "microbial oil" or "oil" mean, unless otherwise specifically 3tated, unmodified oil.
"Microbial oils" or "single cell oils" are those oils 20 naturally produced by microorganisms during their lifespan. Such oils can be high in long chain PUFAs.
The applicant has discovered that certain of these oils, when blended with other microbial oils, fish oils, vegetable oils, or any combination thereof, can 25 produce a composition useful for dietary, pharmaceutical or cosmetic purposes.
Various microbial oils, for example, can be obtained by, for example, the processes disclosed in above-referenced W0 91/14427, W0 91/11918, EP 322,227 30 ~Yamada et al., Suntory) or C~n~l;an Patent Application No. 2,102,273 (W0 92/13086).
,~
It i8 to be under~tood that the present invention encompasses the use of a ~ingle-microbial oil containing at least two desirable PUFAs, such as ARA
and DHA. The oils specifically disclosed and utilized herein, however, each contain a single desirable PUFA.
Any non-toxic, PUFA-containing microbial oil can be used in the present invention. The most preferred microbial oils are those rich in an omega-3 or omega-6 PUFA, especially DHA, G~A or ARA. These PUFAs typically are missing from, or are inadequately provided in, dietary supplements such as infant formulas or baby food. "Infant formula" as used herein means an enteral nutritional product which can be substituted for human brea~t milk in feeding infants and typically is composed of a desired percentage of fat mixed with desired percentages of carbohydrates and proteins in an aqueous solution. Frequently micronutrients, such as trace metals and vit~m; nc or other desired additives are present. Examples of such micronutrients and other additives are disclosed by Cl~n~;n;n et al., U.S. Patent No. 4,670,285.
In the present invention, types of oils from different microbes can be mixed together to obtain a desired composition. Alternatively, or additionally, PUFA-containing microbial oil can be blended with fish oil, vegetable oil or a mixture of both to obtain a desired composition.
An objective in mixing the oils is to obtain an additive which will provide an infant formula with a desired omega-3 and omega-6 PUFA composition similar to that found in breast milk. While the proportion of the desired fatty acids in a microbial oil can vary, this ~, ., .
,~", C
proportion can easily be determined and the amount of oil adjusted to provide the desired amount of PUFA.
Similarly, the percentage of desired PUFA in fish oil or vegetable oils can easily be determined and the amount of the oil to be added can be adjusted as necessary to achieve the desired results.
"Fish oils" are those oils obtained from fish.
Such oils typically contain DHA in amounts ranging from 3% to about 20%. Typically, however, fish oils also contain EPA which depresses the production of ARA in the body. The addition of a microbial oil containing high levels of ARA to fish oil-containing compositions substantially overcomes that problem.
"Vegetable oil" includes all those oils from plants which contain PUFAs. Typically, vegetable oils do not contain long chain ~UF~s (PUFAs at least 20 carbons long), which is why animal organ oils are usually characterized as the source of PUFAs. Thus, vegetarians, especially vegetarian mothers, can have a diet containing inadequate amounts of PUFAs. Vegetable oils known to contain PUFAs may contain GLA. GLA is a Cl~:3 omega-6 PUFA. Such oils include black currant seed oil, borage oil and primrose oil. While GLA is the metabolic precursor to ARA, the process of conversion is very slow, re~uiring the participation of the enzyme ~6-desaturase. This enzyme is present in humans in very low levels. Burre, et al., Lipids, 25:354-356 (1990). Thus, it would be preferable to provide the body with ARA rather than its precursor, GLA.
Methods for isolating vegetable oils are known to those of skill in the art and do not comprise a part of the present invention. Additionally, certain ~ungi 2 7 ~
produce PUFA-containing oils. For example, Mucor species produce a GLA-cont~-n;ng oil.
DHASCO, defined herein as docosahexaneoic acid-containing single cell oill can be obtained, for example, from Crypthecodinium cohnii as disclosed in above-referenced WO 91/11918. DHA is a C22:6 omega-3 long chain PUFA.
EPASCO, defined herein as eicosapentaneoic acid-containing single cell oil, can be obtainedl for example, from Nitzschia alba as disclosed in above-referenced WO 91/144270 EPA i9 a C20:5 omega-3 long chain PUFA.
ARASCO, defined herein as arachidonic acid-containing single cell oil, can be obtained from species such as Pythium insidiosum, or Mortierella alpina, as described in Canadian Patent Application No.
2,101,273 (WO 92/13086). ARA is a C20:4 omega-6 long chain PUFA.
Another aspect of the invention discloses a process for supplementing or altering the composition of commercially available infant formula ~o as to provide them with a PUFA composition more nearly like that typically contained in human breast milk.
"Typical" as used herein refers to the average amounts of PUFAs measured. One of the advantages of the present invention is that, if desired, a nursing mother choosing to switch to formula can have her breast milk analyzed for PUFA content. Then, an additive for a commercially available formula which will supply comparable amounts of PUFAs can be specifically designed. Long chain PUFA-conta;n;ng microbial oils from at least two microorganisms can be obtained and blended together to provide the desired composition.
The blend then can be added to an infant formula.
Preferably, an amount of the blend effective to provide ~ .
210127~
WO92/12711 PCT/US92/~522 an amount of the desired PUFAs substantially similar to that found in human breast milk will be provided.
Typically, human breast milk contains from about 0.5 to 0.6% of its fatty acid content as ARA, from about 0.15 to about 0.36% of its fatty acid content as DHA and from about 0.03 to about 0.13% of its fatty acid content as EPA. Thus, a preferred ratio of ARA:DHA:EPA is from about 5:l:l to about 20:l0:l respectively. Amounts of oils providing approximately these ratios of PUFAs can be determined without undue experimentation by those of skill in the art.
In a preferred embodiment, the microbial oils include ARASCO and DHASCO and EPASCO or any combination thereof. It is also preferred to use oil from microbes of the genera Mortierella, Pythium, Crypthecodini~m, and Nitzschia or any combination thereof. Particularly preferred species from these genera are M. alpina, P.
insidiosum, C. cohnii and N. alba. This preferred embodiment would provide an acceptable alternative for vegetarians, including breast-feeding or pregnant vegetarian women.
If desired, fish oil can be blended, or mixed, with any combination of, or individual, microbial oil to produce a composition which, when subsequently added to infant formula will alter the PUFA content thereof in a desirable manner. Such a composition would not be suitable for a strict vegetarian intake. A preferred fish oil is specially processed Menhaden Oil (produced by Zapata Hayne, Inc.) which typically contains about 9% DHA. Of course, other fish oils also can be used.
When DHASCO is to be blended with ARASCO, and no other PUFA-containing oils are to be utilized, it is desirable to blend sufficient amounts of the oils to provide from about l to about 5 parts DHA with from WO92/12711 2 1 0 1 2 7 4 PCT/US92/~522 ~....
about 2 to about 12 parts ARA. A most preferred ratio of DHA to ARA is l:3 respectively.
As another example, Menhaden fish oil, as noted above, typically contains about 9~ by weight DHA.
ARASCO typically contains about 20 - 40% by weight ARA.
DHASCO typically contains about 25 - 40% by weight DHA.
It has been found that a blend of l part ~e~Aden oil containing about 9% by weight DH~ with l0 parts ARASCO
containing about 33% by weight ARA and 3 parts DHASCO
containing about 35~ by weight DHA, when added to infant formula, causes the infant formula to closely approximate the ARA and DHA content of human breast milk. Other ratios can be readily calculated.
In another embodiment of the present invention is disclosed a process for making a supplement for infant formula or baby food which entails blending a DHA-containing oil with a GLA-containing oil. It is to be understood that, in general, any combination of GLA-, EPA-, ARA- or DHA-containing oils, with or without fish oil, can be used. The source of the GLA can be a vegetable oil, such as primrose, black currant or borage oil, or a microbial oil such as the oil from Mucor javonicus or Mortierella isabellina, for example.
Table l sets forth the GLA composition of such oils.
In a preferred aspect of this embodiment, about l part of Menhaden oil containing about 9% DHA, about 4 parts of GLA-containing oil containing about 18% GLA from black currant seed, and about l part of D~'ASCO
containing about 33% DHA are blended together. Other ratios can be selected as desired.
2 10 1~l~r7 1l WO 92/12711 PCr/US92/00522 --~) I_ o ~r _I x o U O ~ o ~D Lrt ~ ~ ~ ~, ~ .
~ o ~
m ~ n _~ m J ,~
O ~-a) ~ .
O ~ ~
o o~ n O~ :~
I~ O ~ ~
~ _ _ 0 -u~ ~a n o ~ a) F
~_ ~_1 0 ~ _ t-- ~ cn 1' o~ o ~ ~D _t ~ ~ .C --a) ~
C~ 0 ~' o Ul t'-) N CD O O O CO ~ ~_1 a~ N ~ ~ o ol e tn :~ o tr " O
~ ~ w o ~, t tn 0 ~
o ~ ~ N I O e tn c~ O ~n ~~ o e ~ ~
U ~ O ~-1 0--I ~D O ~ D ~ ~a r~ t~
~n ~ 0 _l o o~ ~ t~ ~ CO 1- o ~ _ ~, ~ tn o I
3~ l o ~:5Q
_I ___ _ _ ~, ~ ~I c ~ ~ 0 ~ 3 3-- ~ 3 o o C -~ O _I O --I O _/ N ~ ~ ~r O --I O --I O ~ W
.0 C~ t~ C~ 0 O O N N d' ~n ~
I ~ N N N ~ 3 N
0 ~a I I *1~ :~
~ m ~n ~n o u~ o u~
_I N N' WO92/12711 2 1 0 12~ ~ PCT/US92/~522 A composition including a blend of any combination of the above-described microbial oils with or without either, or both, fish oil and vegetable oil is another aspect of the present invention. While the composition includes any ratios of the oils, the ratios previously described are preferred.
In another preferred embodiment, the composition serves as a nutritional supplement. Typically, such supplements are encapsulated, such as in gelatin capsules. Such capsules provide an easy form of administration to persons having a need for supplementation, such as pregnant or nursing women.
However, parenteral administration is a viable option and in one embodiment the composition comprises the fat component of a total parenteral nutritional formula.
Such formulas are known and commercially available.
As will be understood, the composition of the present invention is particularly useful as a dietary supplement for pregnant or nursing women. Vegetarian women, in particular, may require increased amounts of DHA and ARA, yet have been precluded from obtaining such in the past because the only available sources were animal.
The invention having been previously described in general, reference is now had to the following non-limiting examples for illustrative purposes only.
ExamPles ~xamPle 1. PreParation of P. insidiosum lipid In an 80 liter (gross volume) fermentor, 51 liters of tap water, 1.2 kg glucose, 240 grams of yeast extract and 15 ml of MAZU 210S~ antifoam were combined.
The fermentor was sterilized at 121~C fo- 45 minutes.
An additional 5 liters of condensate water were added ~1~1 2J~q during the sterilization process. The pH was adjusted to 6.2, and approximately 1 liter of inoculum (at a cell density of 5-lOg/l) of PYthium insidiosum tATCC
#28251) then was added. The agitation rate was adjusted to 125 RPM (250 cm/sec tip speed) and the aeration rate was set at 1 SCMF (standard cubic feet per minute). At hour 24 in the operation the aeration rate was increased to 3 SCFM. At hour 28 an additional 2 liters of 50% glucose syrup (1 kg glucose) were added. At hour 50 the fermentor was harvested, resulting in a yield of about 2.2 kg wet weight (approximately 15 g dry weight) per liter. Harvested biomass was squeezed to a high solids cake (50% solids) on a suction filter before freeze drying. The dried biomass was ground with a mortar and pestle and extracted with 1 liter of hexane per 200 grams of dry biomass at room temperature under continuous stirring for 2 hours. The mixture then was filtered and the filtrate evaporated to yield about 5-6 grams of crude oil per 100 grams of dry biomass. The biomass then was reextracted with 1 liter of ethanol per 20 grams of dry biomass for 1 hour at room temperature, filtered, and the solvent evaporated yielding an additional 22 grams of crude oil per 100 grams of dry biomass. The second fraction was predominantly phospholipids whereas the first fraction contained a mixture of phospholipids and triglycerides. The combined fractions produced an oil containing about 30-35% arachidonic acid and no detectable EPA.
Example 2. PreParation of M. alpina liPid Mortierella alpina (ATCC #42430) was grown in a 2 liter shake fl~sk containing 1 liter of tap water and WO92/12711 2 1 0 12 ~ 4 PCT/US92/00522 20 grams of potato dextrose medium. The flask was under constant orbital agitation and was maintained at 25~C for seven days. After harvesting by centrifugation, the biomass was freeze dried yielding about 8 grams of lipid-rich mycelia. The mycelia was extracted using hexane as in example #l and about 2.4g of crude oil resulted. This oil contains about 23%
arachidonic acid.
Example 3 Into a 30-liter working volume STF was loaded a medium of one quarter strength artificial seawater.
Six liters of IO were combined with 18 liters of tap water. The fermentor containing the medium was sterilized and cooled to 28~C. Four hundred ml of concentrated YE (455g/l), 900 ml of glucose syrup t400 g/l) and one liter of inoculum from a seed fermentor containing about 2 x 107 C. cohnii cells/ml or a biomass of 20 g/liter (yielding a final concentration of about 105 cells/ml or a biomass of about 700 mg/liter), were added to the medium. The C. cohnii cells, designated MK8840, were obtained from the American Type Culture Collection as ATCC 40750. Agitation was set at 120 cm/sec tip speed and aeration was set at 1 VVM (30 liters per minute). Additional glucose syrup (900 ml) was added after 30 hours and another 4.2 liters over the next 42 hours. Thus 6 liters of glucose syrup were added in total. Concentrated YE solution ;400 ml) was added at hour 6 and another l.2 liters were added over the next 48 hours until a total of 2.0 liters had been added. To maintain the D.O. at greater than 20~, at 24 hours the agitation tip speed was increased to l50 cm/sec and at 48 hours to 160 cm/sec. At 72 hours, the tip speed was increased to 200 cm/sec and the culture ~lUl~ ~
was permitted to grow for an additional time sufficient to convert the final charge of glucose into cellular oil. The culture was then harvested by centrifugation with the cell pellet retained. The harvested pellet of cells was frozen and dried (lyophilized) to about a 4%
moisture content. Hexane (2.8 liters) was added to the dried biomass and stirred in a glass kettle for l.5 hours at 50~C. A rotary evaporator was used to remove the hexane, producing about l75 g of crude DHA-contAining oil.
Example 4 Into a conventional 30 liter stirred tank fermentor (STF) is added the nutrient medium of Table A, exclusive of the vitAmins, glucose and silicate. The fermentor is equipped with a Rushton-type turbine agitator. The STF and the medium are sterilized. After cooling the medium to about 30~C, the vitamins are added, followed by the addition of sufficient amounts of 40% glucose syrup to provide a glucose concentration of about 80 g/l. Concentrated sodium metasilicate pentahydrate (l00 g/l) is then added to provide a total silicate concentration of about 200 mg/l. Next, the inoculating amount of culture of N. alba cells obtained from the American Type Culture Collection as ATCC 40775, is added in an amount approximately equal to 5% of the total volume of the fermentor, e.g. l.5 liters/30 liters. Agitation is commenced with the tip speed set to 85-90 cm/sec and air sparglng at 1 VVM started. Over about l6 hours an additional charge of concentrated metasilicate (0.53 liters) is added and the agitation speed increased to 126 cm/sec. Over about the next 24 hours, more concentrated silicate (0.83 liters) is added.
Agitation speed again is increased to about 180-185 cm/sec. Over about the next 3 hours an additional 0.15 liters of concentrated metasilicate is added. Thus, the total amount of metasilicate added is about 156 grams or about 1.6 liters of concentrated solution. At about 48 hours additional glucose (about 5 liters) is added, for a total glucose addition of about 4.8 Kg or about 12 liters of 40~ glucose syrup. The culture is permitted to grow for an additional 16 hours, maintaining the agitation speed and aeration rate.
Then, the fermentor is harvested using a Sharples continuous flow centrifuge producing a biomass density of approximately 45-48 grams dry weight per liter. The resulting pellet, about 20-38% solids, is removed and frozen to about -20~C. A vacuum tray drier is used to le...ove water from the pellet. The single cell oil pellet then is extracted with hexane. The hexane subsequently is removed by distillation leaving the extracted single cell oil.
W092/12711 2 1 0 1 2 7 ~ PCT/US92/00522 Table A
GROWTH MEDIUM COMPOSITION
Ingredients needed for 2x30L Fermentors and 2x350L
Fermentors.
Total ReciPe 3OL-Batch 35OL-Batch l9g/L I.O. (Instant Ocean~)570g 6.65Kg 3g/L NaNO3 90g 1.05Kg 0.5g/L NaH2PO4-H20 15g 175g 0.2g/L Na2SiO3-5H20 6g 70g 6ml/L f/2 TM (trace metals)180ml 2.lL
60mg/L H3BO3 1.8g 21g 6mg/L Na2SeO3 180mg 2.lg lOmg/L NaF 300mg 3.5g 40mg/L SrCl2-6H20 1.2g 14g 150mg/L KBr 4.5g 52.5g 0.5g/L KCl 15g 175g 2ml/L B6 TM (trace metals)60ml 700ml After Sterilization O.lml/L of O.lmg/ml Bl2 3ml 35ml O.lml/L of O.lmg/ml Biotin3ml 35ml 2ml/L of lmg/ml Thiamine HCl60ml 700ml Glucose: (1) Start with 80g/L 6L 70L
(40% stock solution) (2) Add another 40g/1 31 35L
1 and 2 (additional 6 liters on day 2) Silicate: Add 60ml/liter of'.8L 21L
lOOg/liter stock solution add additional amounts of stock solution over 48 hours WO92/12711 2 1 0 1 2 7 ~ PCT/US92/00522 Example 5. PreParation of Oil Mix #l and addition to infant formula.
The first mixture represents a totally vegetarian source of an arachidonic and docosahexaenoic acid supplement. This supplement would be considered acceptable to persons restricted to a vegetarian diet.
Sanders et al. (Amer. J. Clin. Nutr. 31:805; 1978) have reported that the DHA levels in the breast milk of vegetarian mothers are depressed. Enteral supplementation of a blend of DHA single cell oil and ARA single cell oil will elevate the serum and, hence, breast milk levels of DHA to that of omnivorous mothers. This blend is prepared by mi xi ng one part DHASCO containing about 35% DHA (obtained from Crypthecodinium cohnii as described in Example 3) with three parts ARASCO containing about 33% ARA (obtained from Pythium insidiosum as described in Example 1).
The resulting mixture, or blend, has the fatty acid composition shown in Table 2. The blend is mixed in a ratio of one part blend to forty parts of the oils regularly in infant formula, typically about 2.8 - 3.0 grams per 100 ml of formula. At a normal fat content of 30g fat per liter of Similac~ infant formula, this corresponds to the addition of 750 mg per liter of prepared formula. This supplement provides ARA and DHA
levels equivalent to human breast milk.
WO 92/12711 21~' ~ 2 7 I PCI/US92/00522 .~
R
o U~
O rY
-I E3 d' U ~ N O _I N O N O O ~ N O~ ~') O _~ N O~
o r-- 0~ 0 N a~ CO O ~D O _I ~ ~ u') O ~ N N _I
;1, _~~.... ~...................... .
'~ d~DOO_~OOOOOOOO
~: Q
,i _~ #
o X
CD ~ 1~ cn ~ ~ L~ I O O O O O O O O N
~'1+ ~ O t~ N Cr~ I O O ~ O O O O N
'a ~a O 0 1' 0 N 0~ 1' 0 O O o o o o o o o o _~ ~ N _I
_I ~
~~1 0 O
a _~
o o~
C: ........ I~IIIIIItI
O ~D O N O 1' 0 1 0 1 ,~ C ~ t~
o #
-1 X o ~ u~ X o c~ o ~ o ~) ~~1 0 ~D O CQ O er N N 11') 1 1 1 ~
._~ ~................ .III.IIII.
0 o ~ r~ ~-- o ~-- t~ N ~ ~ C~
O
E~ O
o t_) O
~ O ~-.,1 ~- ~
O--I
+ ~ C
+
~--1 ~ O O _1 0 H N ~ ~
.~ ~ o -. -- -- -- -- -- -- -- -. -- -- --t~ ta ~- N ~ ~ \ CO CO CO O O O O O Nl N N ~
~4 CC1 ~ I N N N N N N N N N
ul o ~r) o ~ _I ~
21Q12~4 WO92/12711 PCT/US92/~522 ~xamPle 6. PreParation of Oil Nix #2 and addition to infant formula.
This mixture represents a totally vegetarian source of long chain PUFAs and would be considered acceptable to persons restricted to a vegetarian diet.
This blend is prepared by mixing three parts DHASCO
containing about 35% DHA (obtained from Crypthecodinium cohnii as described in Example 3) with ten parts ARASCO
containing about 33~ ARA (obtained from Pythium insidiosum as described in Example 1) and five parts EPASO containing about 5~ EPA (obtained from N. alba as described in Example 4). The resulting mixture, or blend, has the fatty acid composition shown in Table 3.
The blend is mixed in a ratio of one part blend to thirty parts of the oils regularly in infant formula.
At a normal fat content of 30g fat per liter of Similac~ infant formula, this would correspond to the addition of one gram per liter of prepared formula.
This supplement provides ARA, DHA and EPA levels equivalent to human breast milk.
WO 92/12711 ~ 7 ~ PCI/US92/00522 ~n w .. 20 o ..
o U~
o ,, _I
.,~
E3 ~ U ~ o _~ ~ o ~ o o _I ~ o~ ~ ~ ~ ~ o~
~ CO O ~D O ~ o ~ ~ ~ _ .,, ~
o u~ er~oo_Ioooooooo a ,Q
#
x o~_Ioo_Io~oooo o ~ U~ ~ ~ ~ o o o~ I o o ~ o o o o ~
+
O O 1' 0 ~ 01' 00000000000 _I
~.
_I
~_1 ~
o ~
a ~
o O co r~ 7 o ~ o~ _~
~a ~ ............. , .,,,,,,,, _~ O ~D O ~ O r~ O I O I
Q
w o C~l #
~~ X o ~
.~l.............. ll ... l .l .
O ~ D O ~ U~ _I ~ I I O C~ O I O I
o ~
~ ~~
o ~ o ~ ~o ~-~ ~ o o ~
~-1 + ~ ~ C
~+
3 ~ o o ~ o 5~ ~ o O O O O O
o Ul C
~ _I
WO92/12711 2 1 Q i 2 7 4 PCT/US92/00522 2l Exam~le 7. PreParation of Oil Mix ~3 and addition to infant formula.
This mixture is a blend of ARASCO with fish oils.
Oil mixture #3 is prepared by adding one part specially processed Menhaden Oil (Zapata Hayne Inc.) containing about 9% DHA to one part of ARASCO, obtained from Pythium insidios~m as described previously containing about 33% ARA. The resultant fatty acid composition is shown in Table 4. This blend is mixed in a ratio of one part blend to thirty parts of the oils regularly in infant formula. At a normal fat content of 30 g fat per liter of infant formula, this corresponds to the addition of 1 gram per liter of prepared formula. This supplement provides ARA and DHA levels equivalent to human breast milk, but the EPA levels are about eight-fold higher than those in breast milk.
WO 92/12711 . PCI /US92/00522 .
.~
.. .
_I
o o ._ O O _I ~ O ~ O O _t ~ ~ ~ O
r--O~ CO ~ O~ CD O ~D O ~ O ~ ~ ~ ~
_~ J~..................
DOO_IOOOOOOOO
~, ~
~q C ~
~_1 #
_~ X
.~ .,1 o + U7 ~~ ~ ~ O ~ O O r--u~ O O ~r Ln ~ ~ o U~ o o~ _I ~ o o U~ ~ o o o ,~ 0..................
~oot'o~o~ooooooooooo 0 s~
o *
~ ~ .
o s~ _~
o ~,~
C~ ~ C~ ~ ~ O d' O~ _~ O
........ , .,,,,,,,, a) ~ _, o ~D O ~ O t--O I O I
C ~ ~ ~
0 ,.
~U I
,, H ~;
O
#
o ~ oa)oo~DOO~1-- ~
~~ X o ~ U~ Ln ~ o r--O O U~ ~--O O ~ U~
.~,............................... ..
O O ~ ~~ ~ O O 0 1--1--0 0 ~
J
o _I
O o C~ o ~o ~- ~
.~, ... ~ _I
o o--I
+ ~ C
~+
_l ~ ~ ~ ~ ~ ~ er U~ ~ ~ u~ ~D Q
,Q J~ O .. -. -. -- .. -. -. -- -- -- -- -- -- -- -- V~
OOOOO~r~
U~ o ~ o ~- 23 ExamPle 8. PreParation of Oil Mix ~4 and addition to infant formula Oil mixture #4 was developed to utilize GLA in place of arachidonic acid. This blend was prepared by mixing one part specially prepared Menhaden oil containing about 9% DHA (Zapata Hayne Inc.) with four parts black currant seed oil containing about 18% GLA
and one part DHASCO containing about 35~ DHA. The resultant fatty acid composition is shown in Table 5.
This blend is mixed in a ratio of one part blend to forty parts of the oils regularly in infant formula.
At a normal fat content of 30g fat per liter, this would correspond to the addition of 750 mg per liter of prepared formula. This supplement provides EPA and DHA
levels equivalent to human breast milk. The ARA levels are about one tenth the level in human breast milk.
However, the GLA levels are twenty to fifty times higher than the GLA levels in breast milk which typically are minute.
WO 92/12711 2 1 0 1 2 7~ PCl/US92/00~22 24 ~
..
..
_I
w o o .
~.
o o _~ ~ o ~ o o ~ ~ o ~ ~ o~
I' 0'1 ~ ~ 0~ O ~.D O ~ ~D d' U~ O _I ~ N _~
-I ~
~ 0 _I d' ~ ~ ~ ~r ~ o o _I o o o o o o o o _I
.,~ ~
O #
.X
a o ~D O O O ~D O O O _I CO
+ r~ ~ o~ ~ ~ o r~ o o o o o o o _I
~a ~aoo~o~o1~_~oooooooooo U
m o ~o ~a o ........ I . I I I I I I I
O ~D O ~ O r~ O I O I
c~
~C ~
H
o #
~1 X ~ ~ r o ~ ~ o _ I
~-1 ~ ................. I I .. I I ..
u~ o ~ o ~ o o I I o ~ I I o ~--~_I
o U O
~ O
~~1 ~-~ O O ~
~) + CC._ CC~C CCC
-- ~ +
o o _~ o ~ ~ ~ ~ _, ~ ~ ~ In ~ ~r ~ ~D
~. -. -- -- -- -. -- -. -- -- -- -....- -.....
o o o o o Ln o Ir~ O
~ _I ~
WO92/12711 21012 ~ 4 PCT/US92/00522 ExamPle 9. PreParation of Oil Mix #5 and addition to infant for,mula Oil mixture #5 was developed to best approximate the composition of DHA, ARA and EPA of human breast milk. This oil blend was prepared by mixing one part specially prepared Menhaden oil containing about 9% DHA
(zapata Hayne Inc.) with ten parts of ARASCO containing about 33% ARA and three parts DHASCO contAining about 35% DHA. The resultant fatty acid composition is shown in Table 6. This blend is mixed in a ratio of one part blend to forty parts of the oils regularly in infant formula. At a normal fat content of 30g fat per liter of infant formula, this corresponds to the addition of 750 mg per liter of prepared formula. This supplement provides EPA, DHA and ARA levels substantially equivalent to those levels in human breast milk.
?' 26 r~
o _I
..
_, o U~
o o o _l ~ o ~ o o _1 ~ o ~ o ~
I' o~ co o ~o o _I ~ ~ u~ o _1 ~ ~ _I
~-1 ~
~ l~oo_~oooooooo o ~ U~
2 #
X
~a ~O~U~_Ioooc~oooo o ~ ~ ~ ~ o _( o o U~ o o o o +~--oor~o~o~1~ooooooooooo O ~ ~r ~ ,, ~ o ~o o o C~ ~ ~ o ~ C~
........ , .,,,,,,,, C ~ o ~ o ~ o ~--o I o I
C
o U~
o -~ X o ~ o ~
~~1 E3 ~ - ........... ,, ... ,, U~ O ~ ~D CD O t--~ ~ O O I I ~ ~ I I O CO
o ~
- o ~) o ~a O ~.
.~l ~-~
+ c c ~ c c c ~ c c ~
O O _I O ~ ~ r~ ~ ~ ~ ~ er ~ ~ er ~n ~D
O ~- -- -- -- -- -- -- .- .- -. --0 ~ 0 ~ CO 00 0 0 0 0 0 ~ c ~ o ~92/12711 2 1 0 12 ~ 4 PCT/US92/00522 ExamPle lO. PreParation of Oil MIx #6 and addition to infant formula ~, This mixture represents a totally vegetarian source of arachidonic and docosahexaenoic acid. This supplement would be considered acceptable to persons restricted to a vegetarian diet. Sanders et al.
(American J. Clin. Nutr. 31:805; 1978) have reported that the DHA levels in the breast milk of vegetarian mothers are depressed. Enteral supplementation of the blend will elevate the serum and hence breast milk levels of DHA to that of omnivorous mothers. This blend is prepared by mixing one part DHA oil (obtained from Crypthecodinium cohnii as described in Example 3) with five parts ARA oil (obtained from Mortierella alpina as described in Example 2). The resulting mixture has the fatty acid composition shown in Table 7. This blend is mixed in a ratio of one part to thirty-five parts of the oils regularly in infant formula. At a normal fat content of 35 g fat per liter, this would correspond to the addition of l g per liter of prepared formula. This supplement provides ARA and DHA levels equivalent to human breast milk.
~10127~
WO92/12711 PCT/US92/00~22 Table 7. Composition of a blend of DHA oil and ARA oil in proportions of 1:5 by weight.
Infant Formula + Breast Fatt~ Acid Oil Mix #6 Formula Mix #6 Milk 8:0 + 10:00.0 41.8 40.6 1.74 12:0 + 14:04.0 20.7 20.214.95 16:0 16.8 6.8 7.119.82 16:1 0.5 0.2 0.2 3.2 18:0 11.0 2.3 2.5 5.91 18:1 17.8 10.0 10.034.82 18:2 n6 11.1 17.4 17.216.00 18:3 n3 5.2 0.9 0.9 0.62 18:3 n6 4.5 --- 0.1 0.00 20:1 --- 0.1 0.1 1.10 20:2 n6 --- --- 0.0 0.61 20:3 n6 6.0 --- 0.17 0.42 20:4 n6 20.1 --- 0.57 0.59 20:5 n3 0.1 --- 0.01 0.03 22:1 --- --- 0.00 0.10 22:4 n6 2.0 --- 0.06 0.21 22:5 n6 --- --- 0.00 0.22 22:6 n3 6.1 --- 0.18 0.19
MICROBIAL OIL MIXTURES AND USES THEREOF
This invention relates to blends or mixtures of polyunsaturated fatty acid-contA;ni~g microbial oils and to uses thereof. In a specific preferred embodiment, this invention concerns the use of such oils as an additive or supplement for human diets, for example, as an additive to infant formula.
It long has been known that long chain polyunsaturated fatty acids (PUFAs) are essential to the human diet, particularly during periods of rapid tissue growth. Sanders et al, Am. J. Clin. Nutr., 31:805-813 (1978). Certain of these long chain acids, such as arachidonic acid (ARA), cannot be synthesized de novo in humans. Only by metabolizing linoleic acid (LOA), which is converted to gamma linolenic acid (GLA), and then to ARA can the human body produce AR~.
LOA, in turn, is an essential acid which can only be obtained from dietary sources. Additionally, the presence of eicosapentaenoic acid (EPA) in the diet inhibits the metabolic conversion of LOA to ARA.
Carlson, et al., INFORM, 1:306 (1990). ARA and docosahexaneoic acid (DHA) are critical elements of muscle, organ and vascular tissues.
Infancy is the most significant period of rapid growth in a human's life. An infant can increase its body weight by three times or more during its first year of life. Accordingly, it is critical that the infant receive adequate amounts of PUFAs to insure "~,.
proper structural and organ development. Human breast milk contains high levels of PUFAs in which the ratio of ARA to EPA is typically about 20:1. However, many women choose not to breast feed their infants for either part or all of the first year of the infant's life.
A~ recognized by Cl~n~ln;n et al., U.S. Patent 4,670,285, available infant formulas are deficient in long chain (C20 and C22) PUFAs. Cl~n~;n;n et al.
disclose an infant formula prepared from a blend of vegetable oil and egg yolk lipid and/or fish oil which can provide a total fat composition comparable to that of human breast milk. A preferable composition comprise~ from 75 to 95 parts by weight egg yolk and 5 to 25 part~ vegetable oil. This composition iB the entire lipid content of the infant formula and it is not economical to prepare. Additionally, the infant formula disclosed by Cl~n~; n; n et al. results in an EPA
level which i~ 16 times higher than the level of EPA in human breast milk and an ARA level which i8 only one quarter that of brea~t milk.
DE 3603000A1 (Milupa) discloses a computer profile of a highly polyunsaturated acid fat mixture and discusses the use of such a mixture to produce infant formulas. Sources of the fatty acids are listed as certain types of macroalgae (i.e. seaweed), fish oil, organ fats from beef and pork, and highly refined egg yolk oil. In addition to DHA from fish oil, a potential ~ource of DHA and ARA i9 said to be macroalgae, but only of the seaweed types. There is no suggestion to use microbe~ of any type, much less microbial oil.
Methods of producing microbial oils are disclosed in the following references WO/91/14427, published 2 7 ~
~_ 3 October 3, 1991 discloses the production of eicosapentaneoic acid-cont~;n;ng single cell oils (EPASCO). WO 91/11918, published August 22, 1991 discloses the production of docosahexaneoic acid-containing single cell oil (DHASCO). C~nA~;an PatentApplication No. 2,101,273 filed January 22, 1992 (WO
92/13086 published August 6, 1992) relates to the production of arachidonic acid-containing single cell oil (ARASCO). EP 322,227 also discloses a microbial oil production system. None of these references teach the use of blends containing unmodified microbial oils a~ a dietary supplement, or the use of a blend of microbial oils as an additive to exi~ting infant formula to provide that formula with a long chain PUFA
composition similar to breast milk.
Accordingly, it is an object of the present invention to provide a PUFA-enriched additive, the composition of which when added to commercial infant formula will provide desired long chain PUFAs in amounts comparable to the amounts of those PUFAs found in human breast milk.
It is an additional object of the present invention to provide an economical method of producing the above-described composition.
These, and other, objects are satisfied by the present invention as described herein.
Sllmm~ry of the Tnv~nt;on Thi~ invention relates to the use of microbial oils which contain long chain polyunsaturated fatty acids. Additionally, in various embodiments, fish oil and/or vegetable oils can be blended with such microbial oils to form desired compositions. The ~,~
7 ~ ~
_ 4 compositions can be used a~ dietary supplements, particularly as additives for infant formula, as well as for pharmaceutical and cosmetic applications.
The invention also relates to economically viable 5 processes for altering the long chain polyunsaturated fatty acid composition of infant formula and/or baby food. Preferably, the altered composition resembles that of human breast milk.
10Detailed Description of the Preferred ~mhotl;mF~nt of t h ~? TnV~nt;~n ~ roadly stated, the present invention concerns blends, or mixtures, cont~;n;ng unmodified microbial oils. As used herein, "unmodified" mean~ not 15 chemically or covalently altered. It will be understood that throughout this specification references to "microbial oil" or "oil" mean, unless otherwise specifically 3tated, unmodified oil.
"Microbial oils" or "single cell oils" are those oils 20 naturally produced by microorganisms during their lifespan. Such oils can be high in long chain PUFAs.
The applicant has discovered that certain of these oils, when blended with other microbial oils, fish oils, vegetable oils, or any combination thereof, can 25 produce a composition useful for dietary, pharmaceutical or cosmetic purposes.
Various microbial oils, for example, can be obtained by, for example, the processes disclosed in above-referenced W0 91/14427, W0 91/11918, EP 322,227 30 ~Yamada et al., Suntory) or C~n~l;an Patent Application No. 2,102,273 (W0 92/13086).
,~
It i8 to be under~tood that the present invention encompasses the use of a ~ingle-microbial oil containing at least two desirable PUFAs, such as ARA
and DHA. The oils specifically disclosed and utilized herein, however, each contain a single desirable PUFA.
Any non-toxic, PUFA-containing microbial oil can be used in the present invention. The most preferred microbial oils are those rich in an omega-3 or omega-6 PUFA, especially DHA, G~A or ARA. These PUFAs typically are missing from, or are inadequately provided in, dietary supplements such as infant formulas or baby food. "Infant formula" as used herein means an enteral nutritional product which can be substituted for human brea~t milk in feeding infants and typically is composed of a desired percentage of fat mixed with desired percentages of carbohydrates and proteins in an aqueous solution. Frequently micronutrients, such as trace metals and vit~m; nc or other desired additives are present. Examples of such micronutrients and other additives are disclosed by Cl~n~;n;n et al., U.S. Patent No. 4,670,285.
In the present invention, types of oils from different microbes can be mixed together to obtain a desired composition. Alternatively, or additionally, PUFA-containing microbial oil can be blended with fish oil, vegetable oil or a mixture of both to obtain a desired composition.
An objective in mixing the oils is to obtain an additive which will provide an infant formula with a desired omega-3 and omega-6 PUFA composition similar to that found in breast milk. While the proportion of the desired fatty acids in a microbial oil can vary, this ~, ., .
,~", C
proportion can easily be determined and the amount of oil adjusted to provide the desired amount of PUFA.
Similarly, the percentage of desired PUFA in fish oil or vegetable oils can easily be determined and the amount of the oil to be added can be adjusted as necessary to achieve the desired results.
"Fish oils" are those oils obtained from fish.
Such oils typically contain DHA in amounts ranging from 3% to about 20%. Typically, however, fish oils also contain EPA which depresses the production of ARA in the body. The addition of a microbial oil containing high levels of ARA to fish oil-containing compositions substantially overcomes that problem.
"Vegetable oil" includes all those oils from plants which contain PUFAs. Typically, vegetable oils do not contain long chain ~UF~s (PUFAs at least 20 carbons long), which is why animal organ oils are usually characterized as the source of PUFAs. Thus, vegetarians, especially vegetarian mothers, can have a diet containing inadequate amounts of PUFAs. Vegetable oils known to contain PUFAs may contain GLA. GLA is a Cl~:3 omega-6 PUFA. Such oils include black currant seed oil, borage oil and primrose oil. While GLA is the metabolic precursor to ARA, the process of conversion is very slow, re~uiring the participation of the enzyme ~6-desaturase. This enzyme is present in humans in very low levels. Burre, et al., Lipids, 25:354-356 (1990). Thus, it would be preferable to provide the body with ARA rather than its precursor, GLA.
Methods for isolating vegetable oils are known to those of skill in the art and do not comprise a part of the present invention. Additionally, certain ~ungi 2 7 ~
produce PUFA-containing oils. For example, Mucor species produce a GLA-cont~-n;ng oil.
DHASCO, defined herein as docosahexaneoic acid-containing single cell oill can be obtained, for example, from Crypthecodinium cohnii as disclosed in above-referenced WO 91/11918. DHA is a C22:6 omega-3 long chain PUFA.
EPASCO, defined herein as eicosapentaneoic acid-containing single cell oil, can be obtainedl for example, from Nitzschia alba as disclosed in above-referenced WO 91/144270 EPA i9 a C20:5 omega-3 long chain PUFA.
ARASCO, defined herein as arachidonic acid-containing single cell oil, can be obtained from species such as Pythium insidiosum, or Mortierella alpina, as described in Canadian Patent Application No.
2,101,273 (WO 92/13086). ARA is a C20:4 omega-6 long chain PUFA.
Another aspect of the invention discloses a process for supplementing or altering the composition of commercially available infant formula ~o as to provide them with a PUFA composition more nearly like that typically contained in human breast milk.
"Typical" as used herein refers to the average amounts of PUFAs measured. One of the advantages of the present invention is that, if desired, a nursing mother choosing to switch to formula can have her breast milk analyzed for PUFA content. Then, an additive for a commercially available formula which will supply comparable amounts of PUFAs can be specifically designed. Long chain PUFA-conta;n;ng microbial oils from at least two microorganisms can be obtained and blended together to provide the desired composition.
The blend then can be added to an infant formula.
Preferably, an amount of the blend effective to provide ~ .
210127~
WO92/12711 PCT/US92/~522 an amount of the desired PUFAs substantially similar to that found in human breast milk will be provided.
Typically, human breast milk contains from about 0.5 to 0.6% of its fatty acid content as ARA, from about 0.15 to about 0.36% of its fatty acid content as DHA and from about 0.03 to about 0.13% of its fatty acid content as EPA. Thus, a preferred ratio of ARA:DHA:EPA is from about 5:l:l to about 20:l0:l respectively. Amounts of oils providing approximately these ratios of PUFAs can be determined without undue experimentation by those of skill in the art.
In a preferred embodiment, the microbial oils include ARASCO and DHASCO and EPASCO or any combination thereof. It is also preferred to use oil from microbes of the genera Mortierella, Pythium, Crypthecodini~m, and Nitzschia or any combination thereof. Particularly preferred species from these genera are M. alpina, P.
insidiosum, C. cohnii and N. alba. This preferred embodiment would provide an acceptable alternative for vegetarians, including breast-feeding or pregnant vegetarian women.
If desired, fish oil can be blended, or mixed, with any combination of, or individual, microbial oil to produce a composition which, when subsequently added to infant formula will alter the PUFA content thereof in a desirable manner. Such a composition would not be suitable for a strict vegetarian intake. A preferred fish oil is specially processed Menhaden Oil (produced by Zapata Hayne, Inc.) which typically contains about 9% DHA. Of course, other fish oils also can be used.
When DHASCO is to be blended with ARASCO, and no other PUFA-containing oils are to be utilized, it is desirable to blend sufficient amounts of the oils to provide from about l to about 5 parts DHA with from WO92/12711 2 1 0 1 2 7 4 PCT/US92/~522 ~....
about 2 to about 12 parts ARA. A most preferred ratio of DHA to ARA is l:3 respectively.
As another example, Menhaden fish oil, as noted above, typically contains about 9~ by weight DHA.
ARASCO typically contains about 20 - 40% by weight ARA.
DHASCO typically contains about 25 - 40% by weight DHA.
It has been found that a blend of l part ~e~Aden oil containing about 9% by weight DH~ with l0 parts ARASCO
containing about 33% by weight ARA and 3 parts DHASCO
containing about 35~ by weight DHA, when added to infant formula, causes the infant formula to closely approximate the ARA and DHA content of human breast milk. Other ratios can be readily calculated.
In another embodiment of the present invention is disclosed a process for making a supplement for infant formula or baby food which entails blending a DHA-containing oil with a GLA-containing oil. It is to be understood that, in general, any combination of GLA-, EPA-, ARA- or DHA-containing oils, with or without fish oil, can be used. The source of the GLA can be a vegetable oil, such as primrose, black currant or borage oil, or a microbial oil such as the oil from Mucor javonicus or Mortierella isabellina, for example.
Table l sets forth the GLA composition of such oils.
In a preferred aspect of this embodiment, about l part of Menhaden oil containing about 9% DHA, about 4 parts of GLA-containing oil containing about 18% GLA from black currant seed, and about l part of D~'ASCO
containing about 33% DHA are blended together. Other ratios can be selected as desired.
2 10 1~l~r7 1l WO 92/12711 PCr/US92/00522 --~) I_ o ~r _I x o U O ~ o ~D Lrt ~ ~ ~ ~, ~ .
~ o ~
m ~ n _~ m J ,~
O ~-a) ~ .
O ~ ~
o o~ n O~ :~
I~ O ~ ~
~ _ _ 0 -u~ ~a n o ~ a) F
~_ ~_1 0 ~ _ t-- ~ cn 1' o~ o ~ ~D _t ~ ~ .C --a) ~
C~ 0 ~' o Ul t'-) N CD O O O CO ~ ~_1 a~ N ~ ~ o ol e tn :~ o tr " O
~ ~ w o ~, t tn 0 ~
o ~ ~ N I O e tn c~ O ~n ~~ o e ~ ~
U ~ O ~-1 0--I ~D O ~ D ~ ~a r~ t~
~n ~ 0 _l o o~ ~ t~ ~ CO 1- o ~ _ ~, ~ tn o I
3~ l o ~:5Q
_I ___ _ _ ~, ~ ~I c ~ ~ 0 ~ 3 3-- ~ 3 o o C -~ O _I O --I O _/ N ~ ~ ~r O --I O --I O ~ W
.0 C~ t~ C~ 0 O O N N d' ~n ~
I ~ N N N ~ 3 N
0 ~a I I *1~ :~
~ m ~n ~n o u~ o u~
_I N N' WO92/12711 2 1 0 12~ ~ PCT/US92/~522 A composition including a blend of any combination of the above-described microbial oils with or without either, or both, fish oil and vegetable oil is another aspect of the present invention. While the composition includes any ratios of the oils, the ratios previously described are preferred.
In another preferred embodiment, the composition serves as a nutritional supplement. Typically, such supplements are encapsulated, such as in gelatin capsules. Such capsules provide an easy form of administration to persons having a need for supplementation, such as pregnant or nursing women.
However, parenteral administration is a viable option and in one embodiment the composition comprises the fat component of a total parenteral nutritional formula.
Such formulas are known and commercially available.
As will be understood, the composition of the present invention is particularly useful as a dietary supplement for pregnant or nursing women. Vegetarian women, in particular, may require increased amounts of DHA and ARA, yet have been precluded from obtaining such in the past because the only available sources were animal.
The invention having been previously described in general, reference is now had to the following non-limiting examples for illustrative purposes only.
ExamPles ~xamPle 1. PreParation of P. insidiosum lipid In an 80 liter (gross volume) fermentor, 51 liters of tap water, 1.2 kg glucose, 240 grams of yeast extract and 15 ml of MAZU 210S~ antifoam were combined.
The fermentor was sterilized at 121~C fo- 45 minutes.
An additional 5 liters of condensate water were added ~1~1 2J~q during the sterilization process. The pH was adjusted to 6.2, and approximately 1 liter of inoculum (at a cell density of 5-lOg/l) of PYthium insidiosum tATCC
#28251) then was added. The agitation rate was adjusted to 125 RPM (250 cm/sec tip speed) and the aeration rate was set at 1 SCMF (standard cubic feet per minute). At hour 24 in the operation the aeration rate was increased to 3 SCFM. At hour 28 an additional 2 liters of 50% glucose syrup (1 kg glucose) were added. At hour 50 the fermentor was harvested, resulting in a yield of about 2.2 kg wet weight (approximately 15 g dry weight) per liter. Harvested biomass was squeezed to a high solids cake (50% solids) on a suction filter before freeze drying. The dried biomass was ground with a mortar and pestle and extracted with 1 liter of hexane per 200 grams of dry biomass at room temperature under continuous stirring for 2 hours. The mixture then was filtered and the filtrate evaporated to yield about 5-6 grams of crude oil per 100 grams of dry biomass. The biomass then was reextracted with 1 liter of ethanol per 20 grams of dry biomass for 1 hour at room temperature, filtered, and the solvent evaporated yielding an additional 22 grams of crude oil per 100 grams of dry biomass. The second fraction was predominantly phospholipids whereas the first fraction contained a mixture of phospholipids and triglycerides. The combined fractions produced an oil containing about 30-35% arachidonic acid and no detectable EPA.
Example 2. PreParation of M. alpina liPid Mortierella alpina (ATCC #42430) was grown in a 2 liter shake fl~sk containing 1 liter of tap water and WO92/12711 2 1 0 12 ~ 4 PCT/US92/00522 20 grams of potato dextrose medium. The flask was under constant orbital agitation and was maintained at 25~C for seven days. After harvesting by centrifugation, the biomass was freeze dried yielding about 8 grams of lipid-rich mycelia. The mycelia was extracted using hexane as in example #l and about 2.4g of crude oil resulted. This oil contains about 23%
arachidonic acid.
Example 3 Into a 30-liter working volume STF was loaded a medium of one quarter strength artificial seawater.
Six liters of IO were combined with 18 liters of tap water. The fermentor containing the medium was sterilized and cooled to 28~C. Four hundred ml of concentrated YE (455g/l), 900 ml of glucose syrup t400 g/l) and one liter of inoculum from a seed fermentor containing about 2 x 107 C. cohnii cells/ml or a biomass of 20 g/liter (yielding a final concentration of about 105 cells/ml or a biomass of about 700 mg/liter), were added to the medium. The C. cohnii cells, designated MK8840, were obtained from the American Type Culture Collection as ATCC 40750. Agitation was set at 120 cm/sec tip speed and aeration was set at 1 VVM (30 liters per minute). Additional glucose syrup (900 ml) was added after 30 hours and another 4.2 liters over the next 42 hours. Thus 6 liters of glucose syrup were added in total. Concentrated YE solution ;400 ml) was added at hour 6 and another l.2 liters were added over the next 48 hours until a total of 2.0 liters had been added. To maintain the D.O. at greater than 20~, at 24 hours the agitation tip speed was increased to l50 cm/sec and at 48 hours to 160 cm/sec. At 72 hours, the tip speed was increased to 200 cm/sec and the culture ~lUl~ ~
was permitted to grow for an additional time sufficient to convert the final charge of glucose into cellular oil. The culture was then harvested by centrifugation with the cell pellet retained. The harvested pellet of cells was frozen and dried (lyophilized) to about a 4%
moisture content. Hexane (2.8 liters) was added to the dried biomass and stirred in a glass kettle for l.5 hours at 50~C. A rotary evaporator was used to remove the hexane, producing about l75 g of crude DHA-contAining oil.
Example 4 Into a conventional 30 liter stirred tank fermentor (STF) is added the nutrient medium of Table A, exclusive of the vitAmins, glucose and silicate. The fermentor is equipped with a Rushton-type turbine agitator. The STF and the medium are sterilized. After cooling the medium to about 30~C, the vitamins are added, followed by the addition of sufficient amounts of 40% glucose syrup to provide a glucose concentration of about 80 g/l. Concentrated sodium metasilicate pentahydrate (l00 g/l) is then added to provide a total silicate concentration of about 200 mg/l. Next, the inoculating amount of culture of N. alba cells obtained from the American Type Culture Collection as ATCC 40775, is added in an amount approximately equal to 5% of the total volume of the fermentor, e.g. l.5 liters/30 liters. Agitation is commenced with the tip speed set to 85-90 cm/sec and air sparglng at 1 VVM started. Over about l6 hours an additional charge of concentrated metasilicate (0.53 liters) is added and the agitation speed increased to 126 cm/sec. Over about the next 24 hours, more concentrated silicate (0.83 liters) is added.
Agitation speed again is increased to about 180-185 cm/sec. Over about the next 3 hours an additional 0.15 liters of concentrated metasilicate is added. Thus, the total amount of metasilicate added is about 156 grams or about 1.6 liters of concentrated solution. At about 48 hours additional glucose (about 5 liters) is added, for a total glucose addition of about 4.8 Kg or about 12 liters of 40~ glucose syrup. The culture is permitted to grow for an additional 16 hours, maintaining the agitation speed and aeration rate.
Then, the fermentor is harvested using a Sharples continuous flow centrifuge producing a biomass density of approximately 45-48 grams dry weight per liter. The resulting pellet, about 20-38% solids, is removed and frozen to about -20~C. A vacuum tray drier is used to le...ove water from the pellet. The single cell oil pellet then is extracted with hexane. The hexane subsequently is removed by distillation leaving the extracted single cell oil.
W092/12711 2 1 0 1 2 7 ~ PCT/US92/00522 Table A
GROWTH MEDIUM COMPOSITION
Ingredients needed for 2x30L Fermentors and 2x350L
Fermentors.
Total ReciPe 3OL-Batch 35OL-Batch l9g/L I.O. (Instant Ocean~)570g 6.65Kg 3g/L NaNO3 90g 1.05Kg 0.5g/L NaH2PO4-H20 15g 175g 0.2g/L Na2SiO3-5H20 6g 70g 6ml/L f/2 TM (trace metals)180ml 2.lL
60mg/L H3BO3 1.8g 21g 6mg/L Na2SeO3 180mg 2.lg lOmg/L NaF 300mg 3.5g 40mg/L SrCl2-6H20 1.2g 14g 150mg/L KBr 4.5g 52.5g 0.5g/L KCl 15g 175g 2ml/L B6 TM (trace metals)60ml 700ml After Sterilization O.lml/L of O.lmg/ml Bl2 3ml 35ml O.lml/L of O.lmg/ml Biotin3ml 35ml 2ml/L of lmg/ml Thiamine HCl60ml 700ml Glucose: (1) Start with 80g/L 6L 70L
(40% stock solution) (2) Add another 40g/1 31 35L
1 and 2 (additional 6 liters on day 2) Silicate: Add 60ml/liter of'.8L 21L
lOOg/liter stock solution add additional amounts of stock solution over 48 hours WO92/12711 2 1 0 1 2 7 ~ PCT/US92/00522 Example 5. PreParation of Oil Mix #l and addition to infant formula.
The first mixture represents a totally vegetarian source of an arachidonic and docosahexaenoic acid supplement. This supplement would be considered acceptable to persons restricted to a vegetarian diet.
Sanders et al. (Amer. J. Clin. Nutr. 31:805; 1978) have reported that the DHA levels in the breast milk of vegetarian mothers are depressed. Enteral supplementation of a blend of DHA single cell oil and ARA single cell oil will elevate the serum and, hence, breast milk levels of DHA to that of omnivorous mothers. This blend is prepared by mi xi ng one part DHASCO containing about 35% DHA (obtained from Crypthecodinium cohnii as described in Example 3) with three parts ARASCO containing about 33% ARA (obtained from Pythium insidiosum as described in Example 1).
The resulting mixture, or blend, has the fatty acid composition shown in Table 2. The blend is mixed in a ratio of one part blend to forty parts of the oils regularly in infant formula, typically about 2.8 - 3.0 grams per 100 ml of formula. At a normal fat content of 30g fat per liter of Similac~ infant formula, this corresponds to the addition of 750 mg per liter of prepared formula. This supplement provides ARA and DHA
levels equivalent to human breast milk.
WO 92/12711 21~' ~ 2 7 I PCI/US92/00522 .~
R
o U~
O rY
-I E3 d' U ~ N O _I N O N O O ~ N O~ ~') O _~ N O~
o r-- 0~ 0 N a~ CO O ~D O _I ~ ~ u') O ~ N N _I
;1, _~~.... ~...................... .
'~ d~DOO_~OOOOOOOO
~: Q
,i _~ #
o X
CD ~ 1~ cn ~ ~ L~ I O O O O O O O O N
~'1+ ~ O t~ N Cr~ I O O ~ O O O O N
'a ~a O 0 1' 0 N 0~ 1' 0 O O o o o o o o o o _~ ~ N _I
_I ~
~~1 0 O
a _~
o o~
C: ........ I~IIIIIItI
O ~D O N O 1' 0 1 0 1 ,~ C ~ t~
o #
-1 X o ~ u~ X o c~ o ~ o ~) ~~1 0 ~D O CQ O er N N 11') 1 1 1 ~
._~ ~................ .III.IIII.
0 o ~ r~ ~-- o ~-- t~ N ~ ~ C~
O
E~ O
o t_) O
~ O ~-.,1 ~- ~
O--I
+ ~ C
+
~--1 ~ O O _1 0 H N ~ ~
.~ ~ o -. -- -- -- -- -- -- -- -. -- -- --t~ ta ~- N ~ ~ \ CO CO CO O O O O O Nl N N ~
~4 CC1 ~ I N N N N N N N N N
ul o ~r) o ~ _I ~
21Q12~4 WO92/12711 PCT/US92/~522 ~xamPle 6. PreParation of Oil Nix #2 and addition to infant formula.
This mixture represents a totally vegetarian source of long chain PUFAs and would be considered acceptable to persons restricted to a vegetarian diet.
This blend is prepared by mixing three parts DHASCO
containing about 35% DHA (obtained from Crypthecodinium cohnii as described in Example 3) with ten parts ARASCO
containing about 33~ ARA (obtained from Pythium insidiosum as described in Example 1) and five parts EPASO containing about 5~ EPA (obtained from N. alba as described in Example 4). The resulting mixture, or blend, has the fatty acid composition shown in Table 3.
The blend is mixed in a ratio of one part blend to thirty parts of the oils regularly in infant formula.
At a normal fat content of 30g fat per liter of Similac~ infant formula, this would correspond to the addition of one gram per liter of prepared formula.
This supplement provides ARA, DHA and EPA levels equivalent to human breast milk.
WO 92/12711 ~ 7 ~ PCI/US92/00522 ~n w .. 20 o ..
o U~
o ,, _I
.,~
E3 ~ U ~ o _~ ~ o ~ o o _I ~ o~ ~ ~ ~ ~ o~
~ CO O ~D O ~ o ~ ~ ~ _ .,, ~
o u~ er~oo_Ioooooooo a ,Q
#
x o~_Ioo_Io~oooo o ~ U~ ~ ~ ~ o o o~ I o o ~ o o o o ~
+
O O 1' 0 ~ 01' 00000000000 _I
~.
_I
~_1 ~
o ~
a ~
o O co r~ 7 o ~ o~ _~
~a ~ ............. , .,,,,,,,, _~ O ~D O ~ O r~ O I O I
Q
w o C~l #
~~ X o ~
.~l.............. ll ... l .l .
O ~ D O ~ U~ _I ~ I I O C~ O I O I
o ~
~ ~~
o ~ o ~ ~o ~-~ ~ o o ~
~-1 + ~ ~ C
~+
3 ~ o o ~ o 5~ ~ o O O O O O
o Ul C
~ _I
WO92/12711 2 1 Q i 2 7 4 PCT/US92/00522 2l Exam~le 7. PreParation of Oil Mix ~3 and addition to infant formula.
This mixture is a blend of ARASCO with fish oils.
Oil mixture #3 is prepared by adding one part specially processed Menhaden Oil (Zapata Hayne Inc.) containing about 9% DHA to one part of ARASCO, obtained from Pythium insidios~m as described previously containing about 33% ARA. The resultant fatty acid composition is shown in Table 4. This blend is mixed in a ratio of one part blend to thirty parts of the oils regularly in infant formula. At a normal fat content of 30 g fat per liter of infant formula, this corresponds to the addition of 1 gram per liter of prepared formula. This supplement provides ARA and DHA levels equivalent to human breast milk, but the EPA levels are about eight-fold higher than those in breast milk.
WO 92/12711 . PCI /US92/00522 .
.~
.. .
_I
o o ._ O O _I ~ O ~ O O _t ~ ~ ~ O
r--O~ CO ~ O~ CD O ~D O ~ O ~ ~ ~ ~
_~ J~..................
DOO_IOOOOOOOO
~, ~
~q C ~
~_1 #
_~ X
.~ .,1 o + U7 ~~ ~ ~ O ~ O O r--u~ O O ~r Ln ~ ~ o U~ o o~ _I ~ o o U~ ~ o o o ,~ 0..................
~oot'o~o~ooooooooooo 0 s~
o *
~ ~ .
o s~ _~
o ~,~
C~ ~ C~ ~ ~ O d' O~ _~ O
........ , .,,,,,,,, a) ~ _, o ~D O ~ O t--O I O I
C ~ ~ ~
0 ,.
~U I
,, H ~;
O
#
o ~ oa)oo~DOO~1-- ~
~~ X o ~ U~ Ln ~ o r--O O U~ ~--O O ~ U~
.~,............................... ..
O O ~ ~~ ~ O O 0 1--1--0 0 ~
J
o _I
O o C~ o ~o ~- ~
.~, ... ~ _I
o o--I
+ ~ C
~+
_l ~ ~ ~ ~ ~ ~ er U~ ~ ~ u~ ~D Q
,Q J~ O .. -. -. -- .. -. -. -- -- -- -- -- -- -- -- V~
OOOOO~r~
U~ o ~ o ~- 23 ExamPle 8. PreParation of Oil Mix ~4 and addition to infant formula Oil mixture #4 was developed to utilize GLA in place of arachidonic acid. This blend was prepared by mixing one part specially prepared Menhaden oil containing about 9% DHA (Zapata Hayne Inc.) with four parts black currant seed oil containing about 18% GLA
and one part DHASCO containing about 35~ DHA. The resultant fatty acid composition is shown in Table 5.
This blend is mixed in a ratio of one part blend to forty parts of the oils regularly in infant formula.
At a normal fat content of 30g fat per liter, this would correspond to the addition of 750 mg per liter of prepared formula. This supplement provides EPA and DHA
levels equivalent to human breast milk. The ARA levels are about one tenth the level in human breast milk.
However, the GLA levels are twenty to fifty times higher than the GLA levels in breast milk which typically are minute.
WO 92/12711 2 1 0 1 2 7~ PCl/US92/00~22 24 ~
..
..
_I
w o o .
~.
o o _~ ~ o ~ o o ~ ~ o ~ ~ o~
I' 0'1 ~ ~ 0~ O ~.D O ~ ~D d' U~ O _I ~ N _~
-I ~
~ 0 _I d' ~ ~ ~ ~r ~ o o _I o o o o o o o o _I
.,~ ~
O #
.X
a o ~D O O O ~D O O O _I CO
+ r~ ~ o~ ~ ~ o r~ o o o o o o o _I
~a ~aoo~o~o1~_~oooooooooo U
m o ~o ~a o ........ I . I I I I I I I
O ~D O ~ O r~ O I O I
c~
~C ~
H
o #
~1 X ~ ~ r o ~ ~ o _ I
~-1 ~ ................. I I .. I I ..
u~ o ~ o ~ o o I I o ~ I I o ~--~_I
o U O
~ O
~~1 ~-~ O O ~
~) + CC._ CC~C CCC
-- ~ +
o o _~ o ~ ~ ~ ~ _, ~ ~ ~ In ~ ~r ~ ~D
~. -. -- -- -- -. -- -. -- -- -- -....- -.....
o o o o o Ln o Ir~ O
~ _I ~
WO92/12711 21012 ~ 4 PCT/US92/00522 ExamPle 9. PreParation of Oil Mix #5 and addition to infant for,mula Oil mixture #5 was developed to best approximate the composition of DHA, ARA and EPA of human breast milk. This oil blend was prepared by mixing one part specially prepared Menhaden oil containing about 9% DHA
(zapata Hayne Inc.) with ten parts of ARASCO containing about 33% ARA and three parts DHASCO contAining about 35% DHA. The resultant fatty acid composition is shown in Table 6. This blend is mixed in a ratio of one part blend to forty parts of the oils regularly in infant formula. At a normal fat content of 30g fat per liter of infant formula, this corresponds to the addition of 750 mg per liter of prepared formula. This supplement provides EPA, DHA and ARA levels substantially equivalent to those levels in human breast milk.
?' 26 r~
o _I
..
_, o U~
o o o _l ~ o ~ o o _1 ~ o ~ o ~
I' o~ co o ~o o _I ~ ~ u~ o _1 ~ ~ _I
~-1 ~
~ l~oo_~oooooooo o ~ U~
2 #
X
~a ~O~U~_Ioooc~oooo o ~ ~ ~ ~ o _( o o U~ o o o o +~--oor~o~o~1~ooooooooooo O ~ ~r ~ ,, ~ o ~o o o C~ ~ ~ o ~ C~
........ , .,,,,,,,, C ~ o ~ o ~ o ~--o I o I
C
o U~
o -~ X o ~ o ~
~~1 E3 ~ - ........... ,, ... ,, U~ O ~ ~D CD O t--~ ~ O O I I ~ ~ I I O CO
o ~
- o ~) o ~a O ~.
.~l ~-~
+ c c ~ c c c ~ c c ~
O O _I O ~ ~ r~ ~ ~ ~ ~ er ~ ~ er ~n ~D
O ~- -- -- -- -- -- -- .- .- -. --0 ~ 0 ~ CO 00 0 0 0 0 0 ~ c ~ o ~92/12711 2 1 0 12 ~ 4 PCT/US92/00522 ExamPle lO. PreParation of Oil MIx #6 and addition to infant formula ~, This mixture represents a totally vegetarian source of arachidonic and docosahexaenoic acid. This supplement would be considered acceptable to persons restricted to a vegetarian diet. Sanders et al.
(American J. Clin. Nutr. 31:805; 1978) have reported that the DHA levels in the breast milk of vegetarian mothers are depressed. Enteral supplementation of the blend will elevate the serum and hence breast milk levels of DHA to that of omnivorous mothers. This blend is prepared by mixing one part DHA oil (obtained from Crypthecodinium cohnii as described in Example 3) with five parts ARA oil (obtained from Mortierella alpina as described in Example 2). The resulting mixture has the fatty acid composition shown in Table 7. This blend is mixed in a ratio of one part to thirty-five parts of the oils regularly in infant formula. At a normal fat content of 35 g fat per liter, this would correspond to the addition of l g per liter of prepared formula. This supplement provides ARA and DHA levels equivalent to human breast milk.
~10127~
WO92/12711 PCT/US92/00~22 Table 7. Composition of a blend of DHA oil and ARA oil in proportions of 1:5 by weight.
Infant Formula + Breast Fatt~ Acid Oil Mix #6 Formula Mix #6 Milk 8:0 + 10:00.0 41.8 40.6 1.74 12:0 + 14:04.0 20.7 20.214.95 16:0 16.8 6.8 7.119.82 16:1 0.5 0.2 0.2 3.2 18:0 11.0 2.3 2.5 5.91 18:1 17.8 10.0 10.034.82 18:2 n6 11.1 17.4 17.216.00 18:3 n3 5.2 0.9 0.9 0.62 18:3 n6 4.5 --- 0.1 0.00 20:1 --- 0.1 0.1 1.10 20:2 n6 --- --- 0.0 0.61 20:3 n6 6.0 --- 0.17 0.42 20:4 n6 20.1 --- 0.57 0.59 20:5 n3 0.1 --- 0.01 0.03 22:1 --- --- 0.00 0.10 22:4 n6 2.0 --- 0.06 0.21 22:5 n6 --- --- 0.00 0.22 22:6 n3 6.1 --- 0.18 0.19
Claims (53)
1. A process for supplementing infant formula with DHA and ARA which comprises:
a) preparing an oil blend consisting essentially of a microbial oil enriched in DHA and a microbial oil enriched in ARA, wherein the DHA and ARA are in the form of triglycerides and the oils are blended to provide a ratio of about 2 to 12 parts ARA and about 1 to 5 parts DHA and the oil blend further provides an EPA:ARA ratio of about 1 part EPA to from about 5 to about 20 parts ARA; and b) adding said oil blend to said infant formula in sufficient amounts that the amounts of DHA, ARA and EPA in said formula are comparable to the amounts of DHA, ARA and EPA
in human breast milk.
a) preparing an oil blend consisting essentially of a microbial oil enriched in DHA and a microbial oil enriched in ARA, wherein the DHA and ARA are in the form of triglycerides and the oils are blended to provide a ratio of about 2 to 12 parts ARA and about 1 to 5 parts DHA and the oil blend further provides an EPA:ARA ratio of about 1 part EPA to from about 5 to about 20 parts ARA; and b) adding said oil blend to said infant formula in sufficient amounts that the amounts of DHA, ARA and EPA in said formula are comparable to the amounts of DHA, ARA and EPA
in human breast milk.
2. A process in accordance with claim 1, wherein the ARA-containing oil comprises at least 20% ARA.
3. A process in accordance with claim 1, wherein the DHA-containing oil comprises at least about 25%
DHA.
DHA.
4. A process in accordance with claim 1, wherein the DHA-containing oil and the ARA-containing oil are added to the infant formula to provide a ratio of ARA:DHA ranging from about 3:1 to about 2:1.
5. A process in accordance with claim 4, wherein the DHA-containing oil and the ARA-containing oil are added to the infant formula to provide a ratio of ARA:DHA of about 2:1.
6. A process in accordance with claim 1, wherein said ARA-containing oil is obtained by a process comprising cultivating Pythium insidiosum or Mortierella alpina under conditions which will induce the production of an oil enriched in ARA.
7. A process in accordance with claim 1, wherein said DHA-containing oil is obtained by cultivating a DHA-producing species of Crypthecodinium.
8. A process for supplementing infant formula with DHA and ARA which consists essentially of blending a triglyceride oil enriched in DHA and a triglyceride oil enriched in ARA, wherein the oils are blended to provide an ARA:DHA:EPA ratio of from about 5 parts ARA
and 1 part DHA to about 20 parts ARA and 10 parts DHA
to about 1 part EPA, and adding said blend to infant formula in amounts sufficient to provide the infant formula with DHA, ARA and EPA in amounts comparable to the amounts of DHA, ARA and EPA in human breast milk.
and 1 part DHA to about 20 parts ARA and 10 parts DHA
to about 1 part EPA, and adding said blend to infant formula in amounts sufficient to provide the infant formula with DHA, ARA and EPA in amounts comparable to the amounts of DHA, ARA and EPA in human breast milk.
9. A process for supplementing infant formula with DHA and ARA which consists essentially of:
a) obtaining a microbial oil enriched in DHA and blending it with a microbial oil enriched in ARA, wherein the DHA and ARA are in the form of triglycerides and the oils are blended to provide a ratio of about 2 to 12 parts ARA
and about 1 to 5 parts DHA and the oil blend is free of EPA; and b) adding said oil blend to said infant formula in sufficient amounts that the amounts of DHA
and ARA in said formula are comparable to the amounts of DHA and ARA in human breast milk.
a) obtaining a microbial oil enriched in DHA and blending it with a microbial oil enriched in ARA, wherein the DHA and ARA are in the form of triglycerides and the oils are blended to provide a ratio of about 2 to 12 parts ARA
and about 1 to 5 parts DHA and the oil blend is free of EPA; and b) adding said oil blend to said infant formula in sufficient amounts that the amounts of DHA
and ARA in said formula are comparable to the amounts of DHA and ARA in human breast milk.
10. A composition consisting essentially of a blend of a microbial oil enriched in DHA and a microbial oil enriched in ARA, wherein said DHA and ARA
are in the form of triglycerides and the oils are blended to provide an ARA:DHA:EPA ratio of from about 5 parts ARA and 1 part DHA to about 20 parts ARA and 10 parts DHA to about 1 part EPA.
are in the form of triglycerides and the oils are blended to provide an ARA:DHA:EPA ratio of from about 5 parts ARA and 1 part DHA to about 20 parts ARA and 10 parts DHA to about 1 part EPA.
11. A composition in accordance with claim 10, wherein the microbial oil enriched in DHA comprises at least about 25% DHA.
12. A composition in accordance with claim 10, wherein the microbial oil enriched in ARA comprises at least about 20% ARA.
13. A composition consisting essentially of a blend of a microbial oil enriched in DHA and a microbial oil enriched in ARA, wherein the oils are blended to provide a ratio of about 2 to 12 parts ARA
and about 1 to 5 parts DHA.
and about 1 to 5 parts DHA.
14. A composition in accordance with claim 13, wherein the oils are blended to provide a ratio of ARA:DHA of about 2:1.
15. A composition in accordance with claim 13, wherein the amount of EPA is about one twentieth of less the amount of ARA.
16. A composition in accordance with claim 13, wherein the oil enriched in ARA was produced by cultivating Pythium insidiosum or Mortierella alpina under conditions which will induce the production of an oil enriched in ARA.
17. A composition in accordance with claim 13, wherein the oil enriched in DHA was produced by cultivating a DHA-producing species of Crypthecodinium under DHA-producing conditions.
18. A composition comprising a blend of triglyceride oils, wherein said blend consists essentially of ARA, DHA and EPA in a ratio of about 20:10:1 to about 5.101.
19. A composition comprising a blend of a microbial triglyceride oil enriched in ARA and a microbial triglyceride oil enriched in DHA, wherein the microbial triglyceride oils are provided in amounts to provide a ratio of about 2 to 12 parts ARA and about 1 to 5 parts DHA and said oils further are free of EPA.
20. Infant formula comprising a blend of a microbial oil enriched in DHA and a microbial oil enriched in ARA, wherein the DHA and ARA are in the form of triglycerides and the oils are blended to provide a ratio of about 2 to 12 parts ARA and about 1 to 5 parts DHA, the amount of DHA-containing oil and the amount of ARA-containing oil are sufficient to provide amounts of ARA and DHA comparable to the amounts of DHA and ARA in human breast milk and the formula further comprises EPA in a maximum amount of about one twentieth the amount of ARA.
21. Infant formula consisting essentially of a blend of a microbial oil enriched in DHA and an oil enriched in GLA, wherein the amount of the DHA-containing oil is sufficient to provide an amount of DHA comparable to the amount in human breast milk and the amount of the GLA-containing oil is sufficient to provide GLA in an amount that, upon administration of the formula to an infant, can be converted in the infant's body to an amount of ARA comparable to the amount of ARA obtainable from human breast milk.
22. Infant formula comprising DHA and ARA, wherein the DHA and ARA are in the form of triglycerides, the triglycerides are blended to provide a ratio of about 2 to 12 parts ARA and about 1 to 5 parts DHA, and the amount of DHA-containing triglyceride and the amount of ARA-containing triglyceride are sufficient to provide amounts comparable to the amounts of DHA and ARA in human breast milk, wherein said formula is free of EPA.
23. A process for making a supplement for infant formula consisting essentially of blending a docosahexaneoic acid DHA-containing microbial oil and a gamma linolenic acid GLA-containing oil, and adding an amount of the blended oils to infant formula, such that the amount of DHA provided by the blend is sufficient to provide an amount of DHA comparable to the amount in human breast milk and the amount of GLA provided by the blend is sufficient to provide GLA in an amount that, upon administration of the formula to an infant, can be converted in the infant's body to an amount of arachidonic acid (ARA) comparable to the amount of ARA
obtainable from human breast milk.
obtainable from human breast milk.
24. The process of claim 23, wherein said linolenic acid-containing oil comprises primrose, borage, or black currant seed oil.
25. The process of claim 23, wherein said linolenic acid-containing oil comprises a microbial oil.
26. The process of claim 25, further comprising obtaining said linolenic acid-containing oil from Mucor javonicus or Mortierella isabellina.
27. The process of claim 23, further comprising blending an EPA-containing oil with said DHA-containing microbial oil and said linolenic acid-containing oil.
28. The process of claim 27, wherein said EPA-containing oil comprises fish oil.
29. The process of claim 28, wherein said fish oil comprises about one part, said linolenic acid-containing oil comprises about 4 parts and said DHA-containing oil comprises about 1 part by weight of said blend.
30. The process of claim 28, wherein said fish oil comprises about one part, said linolenic acid-containing oil comprises about 4 parts and said DHA-containing oil comprises about 1 part by weight of said blend.
31. A composition consisting essentially of a blend of a DHA-containing microbial oil and a gamma linolenic acid-containing oil, in a ratio such that adding to infant formula an amount of the blended oil composition sufficient to provide an amount of the DHA
comparable to the amount in human breast milk will provide an amount of GLA in the formula that, upon administration of the formula to an infant, can be converted in the infant's body to an amount of ARA
comparable to the amount of ARA obtainable from human breast milk.
comparable to the amount in human breast milk will provide an amount of GLA in the formula that, upon administration of the formula to an infant, can be converted in the infant's body to an amount of ARA
comparable to the amount of ARA obtainable from human breast milk.
32. The composition of claim 31, wherein said linolenic acid-containing oil comprises primrose, borage, or black currant seed oil.
33. The composition of claim 31, wherein said linolenic acid containing-oil is an oil obtained from a microbe.
34. The composition of claim 33, wherein said microbe comprises Mucor javonicus or Mortierella isabellina.
35. The composition of claim 31, also comprising an EPA-containing oil, wherein the amount of EPA
provided by the blend is less than or equal to one-fifth of the amount of ARA obtained from conversion of GLA in the infant's body.
provided by the blend is less than or equal to one-fifth of the amount of ARA obtained from conversion of GLA in the infant's body.
36. The composition of claim 35, wherein said EPA-containing oil comprises fish oil.
37. The composition of claim 36, wherein said fish oil comprises about one part, said linolenic acid-containing oil comprises about 4 parts and said DHA-containing oil comprises about 1 part by weight of said blend.
38. A composition comprising a blend of an oil containing DHA and an oil containing ARA, wherein DHA
and ARA are in the form of triglycerides and further wherein at least one of the oil containing DHA and the oil containing ARA is a microbial oil and ARA:EPA ratio of the blend is at least 5:1.
and ARA are in the form of triglycerides and further wherein at least one of the oil containing DHA and the oil containing ARA is a microbial oil and ARA:EPA ratio of the blend is at least 5:1.
39. A composition according to claim 38, wherein the oils are blended to provide a ratio of about 2 to 12 parts ARA to about 1 to 5 parts DHA.
40. A composition in accordance with claim 39, wherein the microbial oil containing DHA comprises about 25-40% DHA.
41. A composition in accordance with claim 39, wherein the microbial oil containing DHA comprises at least about 35% DHA.
42. A composition in accordance with claim 39, wherein the microbial oil containing ARA comprises about 20-40% ARA.
43. A composition in accordance with claim 39, wherein the oils are blended to provide a ratio of ARA:DHA of about 2:10
44. The composition according to any one of claims 31 or 39, comprising fish oil blended with at least one microbial oil.
45. The composition of claim 44, wherein the ratio of fish oil to microbial oil is from 1:1 to 1:15.
46. A composition according to claim 38, wherein the oils are blended to provide an ARA:DHA:EPA ratio of from about 5 parts ARA and 1 part DHA to about 20 parts ARA and 10 parts DHA to about 1 part EPA.
47. A composition according to claim 38, comprising a blend of a microbial triglyceride oil containing ARA, and a microbial triglyceride oil containing DHA, wherein the microbial triglyceride oils are blended in amounts to provide a ratio of about 2 to 12 parts ARA to about 1 to 5 parts DHA and said oils further are essentially free of EPA.
48. A nutritional supplement containing the composition of any one of claims 31, 38 or 39.
49. A nutritional supplement according to claim 48, comprising a blend of a microbial oil containing DHA and a microbial oil containing ARA, wherein the oils are blended to provide a ratio of about 2 to 12 parts ARA and about 1 to 5 parts DHA, said oils having ARA and DHA in the form of triglycerides.
50. The nutritional supplement according to claim 48, wherein the supplement is a human nutritional supplement.
51. The nutritional supplement according to claim 48, wherein the human is a baby.
52. The nutritional supplement according to claim 48, wherein the human is a pregnant or nursing woman.
53. A total parenteral nutrition formula containing the composition according to any of claims 31, 38 or 39.
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US64545791A | 1991-01-24 | 1991-01-24 | |
US645,457 | 1991-01-24 |
Publications (2)
Publication Number | Publication Date |
---|---|
CA2101274A1 CA2101274A1 (en) | 1992-07-25 |
CA2101274C true CA2101274C (en) | 1998-12-15 |
Family
ID=24589106
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CA002101274A Expired - Lifetime CA2101274C (en) | 1991-01-24 | 1992-01-22 | Microbial oil mixtures and uses thereof |
Country Status (20)
Country | Link |
---|---|
US (2) | US5374657A (en) |
EP (5) | EP0568606B1 (en) |
JP (1) | JP2731035B2 (en) |
KR (3) | KR100321543B1 (en) |
AT (1) | ATE200619T1 (en) |
AU (1) | AU661297B2 (en) |
BR (1) | BR9205526A (en) |
CA (1) | CA2101274C (en) |
DE (2) | DE69231793T2 (en) |
DK (1) | DK0568606T3 (en) |
ES (1) | ES2157898T3 (en) |
GR (1) | GR3036139T3 (en) |
IL (2) | IL114253A (en) |
MX (1) | MX183638B (en) |
NZ (1) | NZ241359A (en) |
OA (1) | OA10348A (en) |
RU (1) | RU2093996C1 (en) |
SG (1) | SG49307A1 (en) |
WO (1) | WO1992012711A1 (en) |
ZA (1) | ZA92452B (en) |
Families Citing this family (225)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
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 |
US5985348A (en) * | 1995-06-07 | 1999-11-16 | Omegatech, Inc. | Milk products having high concentrations of omega-3 highly unsaturated fatty acids |
US6451567B1 (en) | 1988-09-07 | 2002-09-17 | Omegatech, Inc. | Fermentation process for producing long chain omega-3 fatty acids with euryhaline microorganisms |
US6410281B1 (en) | 1992-07-10 | 2002-06-25 | Omegatech, Inc. | Reducing corrosion in a fermentor by providing sodium with a non-chloride sodium salt |
DE69433713T2 (en) * | 1993-06-09 | 2005-04-07 | Martek Biosciences Corp. | USEFUL METHODS AND PHARMACEUTICAL COMPOSITIONS FOR THE TREATMENT OF NEUROLOGICAL DISEASES |
US20050027004A1 (en) * | 1993-06-09 | 2005-02-03 | Martek Biosciences Corporation | Methods of treating senile dementia and Alzheimer's diseases using docosahexaenoic acid and arachidonic acid compositions |
WO1995009622A1 (en) * | 1993-10-06 | 1995-04-13 | Peptide Technology Limited | Polyunsaturated fatty acids and uses thereof |
ATE183391T1 (en) * | 1993-12-31 | 1999-09-15 | Univ Limburg | USE OF ESSENTIAL FATTY ACID COMPOSITIONS |
CA2150741A1 (en) * | 1994-06-17 | 1995-12-18 | Susan Trimbo | Pediatric lipid emulsion |
JP3419897B2 (en) * | 1994-07-22 | 2003-06-23 | 明治乳業株式会社 | Hypoallergenic formula |
CA2201931C (en) * | 1994-10-05 | 2004-08-10 | Gerhard Kohn | Mixture of phospholipid-containing fats and lcp fatty acids |
US5583019A (en) | 1995-01-24 | 1996-12-10 | Omegatech Inc. | Method for production of arachidonic acid |
AU706613B2 (en) * | 1995-02-27 | 1999-06-17 | Clover Corporation Pty Ltd | Supplement for baby infant formula and a method of delivering that supplement |
AUPN137895A0 (en) * | 1995-02-27 | 1995-03-16 | Clover Corporation Pty Ltd | Composition and method |
US20080175953A1 (en) * | 1995-06-07 | 2008-07-24 | Martek Biosciences Corporation | Process for the Heterotrophic Production of Microbial Products with High Concentrations of Omega-3 Highly Unsaturated Fatty Acids |
US6428832B2 (en) * | 1996-03-26 | 2002-08-06 | Dsm N.V. | Late addition of PUFA in infant formula preparation process |
EP0893953B1 (en) * | 1996-03-26 | 2002-06-05 | Dsm N.V. | Pufa coated solid carrier particles for foodstuff |
US6048557A (en) * | 1996-03-26 | 2000-04-11 | Dsm N.V. | PUFA coated solid carrier particles for foodstuff |
US6255505B1 (en) | 1996-03-28 | 2001-07-03 | Gist-Brocades, B.V. | Microbial polyunsaturated fatty acid containing oil from pasteurised biomass |
DE69724782T3 (en) * | 1996-03-28 | 2015-12-24 | Dsm Ip Assets B.V. | Process for producing granular microbial biomass and obtaining valuable components from microbial biomass |
DK0894142T4 (en) * | 1996-03-28 | 2014-02-24 | Dsm Ip Assets Bv | Microbial oil comprising a polyunsaturated fatty acid and process for producing oil from pasteurized and granulated biomass. |
US20030143659A1 (en) * | 1996-03-28 | 2003-07-31 | Hendrik Louis Bijl | Process for the preparation of a granular microbial biomass and isolation of a compound thereform |
AU2956397A (en) * | 1996-05-15 | 1997-12-05 | Gist-Brocades B.V. | Sterol extraction with polar solvent to give low sterol, high triglyceride, microbial oil |
US5625062A (en) * | 1996-05-29 | 1997-04-29 | Minnesota Mining And Manufacturing Company | Method of making soluble squaraine dyes |
CA2208392C (en) * | 1996-06-21 | 2002-09-03 | University Of Guelph | Method for enriching docosahexaenoic acid in expressed milk of dairy cattle |
DE19724845A1 (en) * | 1996-08-28 | 1998-03-05 | Solvay Pharm Gmbh | Use of complex lipids as stabilizing additives for pharmaceutical preparations of digestive enzyme mixtures |
US5995724A (en) * | 1996-11-01 | 1999-11-30 | Mikkelsen; Carl | Image process system and process using personalization techniques |
PL343902A1 (en) | 1997-02-21 | 2001-09-10 | Abbott Lab | Methods of and compositions for reducing enteritis and colitis morbidity rate |
US6080787A (en) * | 1997-02-21 | 2000-06-27 | Abbott Laboratories | Methods for reducing the incidence of necrotizing enterocolitis |
CN1660065A (en) * | 1997-03-27 | 2005-08-31 | 布里斯托尔-迈尔斯斯奎布公司 | Use of docosahexaenoic acid and arachidonic acid enhancing the growth of preterm infants |
US6051754A (en) * | 1997-04-11 | 2000-04-18 | Abbott Laboratories | Methods and compositions for synthesis of long chain poly-unsaturated fatty acids in plants |
US7745694B1 (en) | 1997-04-11 | 2010-06-29 | Monsanto Technology Llc | Methods and compositions for synthesis of long chain polyunsaturated fatty acids in plants |
US5968809A (en) * | 1997-04-11 | 1999-10-19 | Abbot Laboratories | Methods and compositions for synthesis of long chain poly-unsaturated fatty acids |
US6075183A (en) * | 1997-04-11 | 2000-06-13 | Abbott Laboratories | Methods and compositions for synthesis of long chain poly-unsaturated fatty acids in plants |
US5972664A (en) | 1997-04-11 | 1999-10-26 | Abbott Laboratories | Methods and compositions for synthesis of long chain poly-unsaturated fatty acids |
US5993221A (en) * | 1997-05-01 | 1999-11-30 | Beth Israel Deaconess Medical Center, Inc. | Dietary formulation comprising arachidonic acid and methods of use |
JP2001512029A (en) * | 1997-08-01 | 2001-08-21 | マーテック・バイオサイエンスィズ・コーポレーション | DHA-containing nutritional compositions and methods for their production |
DE19836339B4 (en) | 1998-08-11 | 2011-12-22 | N.V. Nutricia | carbohydrate mix |
WO2000011188A1 (en) * | 1998-08-18 | 2000-03-02 | Metabolix, Inc. | Transgenic microbial polyhydroxyalkanoate producers |
EP2308486A1 (en) * | 1998-10-15 | 2011-04-13 | DSM IP Assets B.V. | Pufa supplements |
US6166231A (en) * | 1998-12-15 | 2000-12-26 | Martek Biosciences Corporation | Two phase extraction of oil from biomass |
AU4030600A (en) * | 1999-03-26 | 2000-10-16 | Martek Biosciences Corporation | Specific binding assay for docosahexaenoic acid |
US6258846B1 (en) * | 1999-06-01 | 2001-07-10 | Drugtech Corporation | Nutritional supplements |
US7112609B2 (en) * | 1999-06-01 | 2006-09-26 | Drugtech Corporation | Nutritional supplements |
GB9916537D0 (en) * | 1999-07-14 | 1999-09-15 | Univ Hull | Culture of microorganisms for the synthesis of a polyunsaturated fatty acid |
NO313076B1 (en) * | 1999-12-28 | 2002-08-12 | Pronova Biocare As | Liquid nutrients and / or nutrients and processes for their preparation |
EP2341126A3 (en) | 2000-01-28 | 2011-10-05 | Martek Biosciences Corporation | Enhanced production of lipids containing polyenoic fatty acids by high density cultures of eukaryotic microbes in fermentors |
US6495599B2 (en) | 2000-04-13 | 2002-12-17 | Abbott Laboratories | Infant formulas containing long-chain polyunsaturated fatty acids and uses therof |
EP1276885B1 (en) | 2000-04-21 | 2007-09-12 | Martek Biosciences Corporation | Trophic conversion of obligate phototrophic algae through metabolic engineering |
EP1780283A1 (en) | 2000-04-21 | 2007-05-02 | Martek Biosciences Corporation | Trophic conversion of obligate photographic algae through metabolic engineering |
AU7302801A (en) | 2000-06-26 | 2002-01-08 | Omegatech Inc | Improved methods of incorporating polyunsaturated fatty acids in milk |
EP1178103A1 (en) * | 2000-08-02 | 2002-02-06 | Dsm N.V. | Purifying crude pufa oils |
KR20010008387A (en) | 2000-11-30 | 2001-02-05 | 이성권 | Method for producing highly pure unsaturated fatty acid using crystallization |
GB2377455A (en) * | 2001-02-09 | 2003-01-15 | Univ Hull | Method of culturing crypthecodinium cohnii |
US20050129739A1 (en) * | 2001-05-14 | 2005-06-16 | Gerhard Kohn | Production and use of a polar lipid-rich fraction containing omega-3 and/or omega-6 highly unsaturated fatty acids from microbes, genetically modified plant seeds and marine organisms |
WO2003017945A2 (en) * | 2001-08-24 | 2003-03-06 | Martek Biosciences Boulder Corporation | Products containing highly unsaturated fatty acids for use by women and their children during stages of preconception, pregnancy and lactation/post-partum |
US7704542B2 (en) * | 2001-09-12 | 2010-04-27 | Xanodyne Pharmaceuticals, Inc. | Vitamin/mineral compositions with DHA |
US20040048926A1 (en) * | 2002-03-15 | 2004-03-11 | Hoffman Dennis Robert | Use of docosahexaenoic acid and arachidonic acid to enhance the visual development of term infants breast-fed up to the age of six months |
EP1515616A4 (en) * | 2002-03-19 | 2005-12-28 | Advanced Bionutrition Corp | Microalgal feeds containing arachidonic acid and their production and use |
BR0309986A (en) * | 2002-05-14 | 2005-03-01 | J Oil Mills Inc | Flavor enhancer, method for enhancing the flavor of vegetable oil and fat, composition of vegetable oil and fat, and, food |
US20050129738A1 (en) * | 2002-06-16 | 2005-06-16 | Lipogen Ltd. | Infant formula supplemented with phospholipids |
JP2005529728A (en) * | 2002-06-18 | 2005-10-06 | マーテック・バイオサイエンシーズ・コーポレーション | Stable emulsion of oil in aqueous solution and process for its production |
EP3111767B1 (en) | 2002-06-19 | 2018-11-21 | DSM IP Assets B.V. | Microbial oil and processes for its processing |
AU2003251557A1 (en) * | 2002-06-28 | 2004-01-19 | Richard C. Theuer | Fat compositions for infant formula and methods therefor |
AU2003251715A1 (en) * | 2002-08-30 | 2004-03-19 | Campina B.V. | Foaming ingredient and products containing the ingredient |
DE60228707D1 (en) * | 2002-09-04 | 2008-10-16 | Nestec Sa | Process for the preparation of an oil containing long-chain polyunsaturated fatty acids from biomass, food, food composition, cosmetic or pharmaceutical composition containing this oil |
US20040209953A1 (en) * | 2002-12-06 | 2004-10-21 | Wai Lee Theresa Siu-Ling | Glyceride compositions and methods of making and using same |
US8084074B2 (en) * | 2003-02-12 | 2011-12-27 | E. I. Du Pont De Nemours And Company | Production of very long chain polyunsaturated fatty acids in oil seed plants |
US7214491B2 (en) * | 2003-05-07 | 2007-05-08 | E. I. Du Pont De Nemours And Company | Δ-12 desaturase gene suitable for altering levels of polyunsaturated fatty acids in oleaginous yeasts |
AU2003902823A0 (en) * | 2003-06-04 | 2003-06-26 | Athol Gillies Turner | Biologically active oils |
CA2532472C (en) * | 2003-06-23 | 2012-04-17 | Nestec S.A. | Infant or follow-on formula |
NZ544726A (en) * | 2003-06-23 | 2009-05-31 | Nestec Sa | Infant or follow-on formula |
CN1842277A (en) * | 2003-06-24 | 2006-10-04 | 堪萨斯大学医学中心 | Infant formula |
KR20060027862A (en) * | 2003-07-09 | 2006-03-28 | 제이-오일 밀스, 인코포레이티드 | Antioxidant fat or oil composition containing long-chain highly unsaturated fatty acid |
EP1645197A4 (en) * | 2003-07-09 | 2008-12-24 | J Oil Mills Inc | Full-bodied taste enhancer containing product of decomposition of long-chain highly unsaturated fatty acid or containing extract therefrom |
TW201119585A (en) * | 2003-11-12 | 2011-06-16 | J Oil Mills Inc | Body taste improver comprising long-chain highly unsaturated fatty acid and/or its ester |
TW200526131A (en) * | 2003-11-12 | 2005-08-16 | J Oil Mills Inc | Method of application of body taste enhancer comprising long-chain highly unsaturated fatty acid and/or its ester |
WO2005092370A1 (en) * | 2004-03-22 | 2005-10-06 | Solvay Pharmaceuticals Gmbh | Oral pharmaceutical compositions of lipase-containing products, in particular of pancreatin, containing surfactants |
EP1597978A1 (en) | 2004-05-17 | 2005-11-23 | Nutricia N.V. | Synergism of GOS and polyfructose |
US8075910B2 (en) * | 2004-05-20 | 2011-12-13 | Pbm Pharmaceuticals, Inc. | Oral compositions comprising edible oils and vitamins and/or minerals and methods for making oral compositions |
US8252769B2 (en) | 2004-06-22 | 2012-08-28 | N. V. Nutricia | Intestinal barrier integrity |
EP1721611A1 (en) * | 2005-04-21 | 2006-11-15 | N.V. Nutricia | Nutritional supplement with oligosaccharides for a category of HIV patients |
AU2005253898B2 (en) * | 2004-06-22 | 2010-02-18 | N. V. Nutricia | Improvement of barrier integrity in hiv patients with fatty acids |
EP1723951A1 (en) * | 2005-04-21 | 2006-11-22 | N.V. Nutricia | Nutritional supplement with oligosaccharides for a category of HIV patients |
EP1634599A1 (en) | 2004-08-20 | 2006-03-15 | N.V. Nutricia | Iimmune stimulatory infant nutrition |
DE102004062141A1 (en) * | 2004-12-23 | 2006-07-06 | Nutrinova Nutrition Specialties & Food Ingredients Gmbh | Process for the preparation of a crude oil from mixtures of microorganisms and plants, the oil thus produced and the specific uses of the thus prepared and optionally additionally refined oil |
AU2006227165B2 (en) | 2005-03-18 | 2011-11-10 | Microbia, Inc. | Production of carotenoids in oleaginous yeast and fungi |
US20060229366A1 (en) | 2005-04-07 | 2006-10-12 | Lifschitz Carlos H | Method for preventing or treating respiratory infections in infants |
MX2007013075A (en) * | 2005-04-21 | 2008-01-11 | Nutricia Nv | Nutritional supplement for hiv patients. |
US7572474B2 (en) | 2005-06-01 | 2009-08-11 | Mead Johnson Nutrition Company | Method for simulating the functional attributes of human milk oligosaccharides in formula-fed infants |
US20060286208A1 (en) * | 2005-06-01 | 2006-12-21 | Nagendra Rangavajla | Methods for producing protein partial hydrolysates and infant formulas containing the same |
EP2447356B1 (en) | 2005-06-07 | 2016-04-20 | DSM Nutritional Products AG | Eukaryotic microorganisms for producing lipids and antioxidants |
US8075934B2 (en) * | 2008-10-24 | 2011-12-13 | Mead Johnson Nutrition Company | Nutritional composition with improved digestibility |
MX300085B (en) * | 2005-07-01 | 2012-06-08 | Martek Biosciences Corp | Polyunsaturated fatty acid-containing oil product and uses and production thereof. |
WO2007005727A2 (en) * | 2005-07-01 | 2007-01-11 | Martek Biosciences Corporation | Microwaveable popcorn and methods of making |
WO2007014896A1 (en) | 2005-07-29 | 2007-02-08 | Solvay Pharmaceuticals Gmbh | Processes for the manufacture of sterilized pancreatin powder |
US11266607B2 (en) * | 2005-08-15 | 2022-03-08 | AbbVie Pharmaceuticals GmbH | Process for the manufacture and use of pancreatin micropellet cores |
US9198871B2 (en) * | 2005-08-15 | 2015-12-01 | Abbott Products Gmbh | Delayed release pancreatin compositions |
US20070166354A1 (en) | 2005-10-26 | 2007-07-19 | Bridget Barrett-Reis | Method of reducing the risk of retinopathy of prematurity in preterm infants |
US7829126B2 (en) * | 2005-10-26 | 2010-11-09 | Abbott Laboratories | Infant formulas containing docosahexaenoic acid and lutein |
US20070166411A1 (en) * | 2005-12-16 | 2007-07-19 | Bristol-Myers Squibb Company | Nutritional supplement containing long-chain polyunsaturated fatty acids |
DE602006015701D1 (en) * | 2005-12-29 | 2010-09-02 | Abl Biotechnologies Ltd | NEW SCHIZOCHYTRIUM LIMACINUM TRUNK, SUITABLE FOR THE PRODUCTION OF LIPIDES AND EXTRACELLULAR POLYSACCHARIDES, AND METHOD THEREFOR |
US20070243307A1 (en) * | 2006-04-11 | 2007-10-18 | Martek Biosciences Corporation | Food Products Comprising Long Chain Polyunsaturated Fatty Acids and Methods for Preparing the Same |
US10072256B2 (en) * | 2006-05-22 | 2018-09-11 | Abbott Products Gmbh | Process for separating and determining the viral load in a pancreatin sample |
EP1886680A1 (en) * | 2006-05-23 | 2008-02-13 | Nestec S.A. | Maternal supplement |
US8221809B2 (en) | 2006-06-22 | 2012-07-17 | Martek Biosciences Corporation | Encapsulated labile compound compositions and methods of making the same |
EP2043970B1 (en) * | 2006-07-05 | 2018-04-18 | Centre National De La Recherche Scientifique (C.N.R.S.) | Iron-copper co-catalyzed process for carbon-carbon or carbon-heteroatom bonding |
US20080026105A1 (en) * | 2006-07-28 | 2008-01-31 | Bristol-Myers Squibb Company | Nutritional formulations containing octenyl succinate anhydride-modified tapioca starch |
MX2009001346A (en) | 2006-08-01 | 2009-04-17 | Ocean Nutrition Canada Ltd | Oil producing microbes and methods of modification thereof. |
WO2008027991A2 (en) * | 2006-08-29 | 2008-03-06 | Martek Biosciences Corporation | USE OF DPA(n-6) OILS IN INFANT FORMULA |
AU2007299896B2 (en) * | 2006-09-18 | 2013-07-11 | The Arizona Board Of Regents, A Body Corporate Of The State Of Arizona Acting For And On Behalf Of Arizona State University | Algal medium chain length fatty acids and hydrocarbons |
WO2008042338A2 (en) | 2006-09-28 | 2008-04-10 | Microbia, Inc. | Production of carotenoids in oleaginous yeast and fungi |
BRPI0810138A2 (en) * | 2007-05-18 | 2014-09-23 | Mead Johnson Nutrition Co | ACIDIFIED LIQUID HUMAN MILK SUPPLEMENT |
EP2025237A1 (en) * | 2007-08-15 | 2009-02-18 | Nestec S.A. | Lecithin and LC-PUFA |
US8148559B1 (en) | 2007-08-31 | 2012-04-03 | Clemson University Research Foundation | Supercritical fluid explosion process to aid fractionation of lipids from biomass |
MX335992B (en) * | 2007-08-31 | 2016-01-07 | Dsm Ip Assets Bv | Polyunsaturated fatty acid-containing solid fat compositions and uses and production thereof. |
US8343753B2 (en) * | 2007-11-01 | 2013-01-01 | Wake Forest University School Of Medicine | Compositions, methods, and kits for polyunsaturated fatty acids from microalgae |
US20090130063A1 (en) * | 2007-11-15 | 2009-05-21 | Solvay Pharmaceuticals Gmbh | Process for separating and determining the viral load in a pancreatin sample |
WO2009096772A1 (en) * | 2008-02-01 | 2009-08-06 | N.V. Nutricia | Composition for stimulating natural killer cell activity |
US20090202672A1 (en) * | 2008-02-11 | 2009-08-13 | Monsanto Company | Aquaculture feed, products, and methods comprising beneficial fatty acids |
CN101259101B (en) * | 2008-04-17 | 2010-06-02 | 湖北福星生物科技有限公司 | Micrometre level arachidonic acid/docosahexaenoic acid emulsion and preparation thereof |
AP2776A (en) | 2008-06-19 | 2013-09-30 | Unilever Plc | Fat containing edible emulsions with iron and zinc |
US8498729B2 (en) | 2008-08-29 | 2013-07-30 | Smp Logic Systems Llc | Manufacturing execution system for use in manufacturing baby formula |
US8986769B2 (en) * | 2008-10-24 | 2015-03-24 | Mead Johnson Nutrition Company | Methods for preserving endogenous TGF-β |
US8425955B2 (en) | 2009-02-12 | 2013-04-23 | Mead Johnson Nutrition Company | Nutritional composition with prebiotic component |
ES2570774T3 (en) * | 2009-04-01 | 2016-05-20 | Nestec Sa | Use of arachidonic acid to reduce the risk of insulin resistance later in life |
EP2295535A1 (en) | 2009-09-11 | 2011-03-16 | Mead Johnson Nutrition Company | Probiotic material |
US20140093614A1 (en) | 2009-09-20 | 2014-04-03 | Mead Johnson Nutrition Company | Probiotic stabilization |
US20110070334A1 (en) * | 2009-09-20 | 2011-03-24 | Nagendra Rangavajla | Probiotic Stabilization |
KR101889561B1 (en) * | 2009-11-03 | 2018-08-17 | 디에스엠 아이피 어셋츠 비.브이. | Composition comprising cells and a polyunsaturated fatty acid having at least 20 carbon atoms(lc-pufa) |
WO2011066419A2 (en) * | 2009-11-25 | 2011-06-03 | Kuehnle Agrosystems, Inc. | Enrichment of process feedstock |
PL2353595T3 (en) | 2010-01-19 | 2016-03-31 | Mjn Us Holdings Llc | Nutritional compensation for western-type diet |
BR112012017831B8 (en) * | 2010-01-19 | 2021-05-25 | Dsm Ip Assets Bv | microbial oil, animal food and biomass comprising said microbial oil |
PE20121729A1 (en) | 2010-01-29 | 2013-01-16 | Abbott Lab | NUTRITIONAL EMULSIONS INCLUDING CALCIUM BETA-HYDROXY-BETA-METHYLBUTYRATE (HMB) |
US9693577B2 (en) | 2010-01-29 | 2017-07-04 | Abbott Laboratories | Method of preparing a nutritional powder comprising spray dried HMB |
WO2011094548A1 (en) | 2010-01-29 | 2011-08-04 | Abbott Laboratories | Aseptically packaged nutritional liquids comprising hmb |
WO2011103514A1 (en) * | 2010-02-18 | 2011-08-25 | Martek Biosciences Corporation | Dha triglyceride emulsions |
US8202425B2 (en) | 2010-04-06 | 2012-06-19 | Heliae Development, Llc | Extraction of neutral lipids by a two solvent method |
US8211309B2 (en) | 2010-04-06 | 2012-07-03 | Heliae Development, Llc | Extraction of proteins by a two solvent method |
US8115022B2 (en) | 2010-04-06 | 2012-02-14 | Heliae Development, Llc | Methods of producing biofuels, chlorophylls and carotenoids |
US8211308B2 (en) | 2010-04-06 | 2012-07-03 | Heliae Development, Llc | Extraction of polar lipids by a two solvent method |
US8308951B1 (en) | 2010-04-06 | 2012-11-13 | Heliae Development, Llc | Extraction of proteins by a two solvent method |
WO2011127127A2 (en) | 2010-04-06 | 2011-10-13 | Arizona Board Of Regents For And On Behalf Of Arizona State University | Extraction with fractionation of oil and co-products from oleaginous material |
US8313648B2 (en) | 2010-04-06 | 2012-11-20 | Heliae Development, Llc | Methods of and systems for producing biofuels from algal oil |
US8273248B1 (en) | 2010-04-06 | 2012-09-25 | Heliae Development, Llc | Extraction of neutral lipids by a two solvent method |
US8475660B2 (en) | 2010-04-06 | 2013-07-02 | Heliae Development, Llc | Extraction of polar lipids by a two solvent method |
KR20130048218A (en) | 2010-04-06 | 2013-05-09 | 헬리아에 디벨롭먼트, 엘엘씨 | Methods of and systems for dewatering algae and recycling water therefrom |
BR112012023328A2 (en) | 2010-05-28 | 2016-08-23 | Mead Johnson Nutrition Co | nutritional compositions |
US20110293784A1 (en) | 2010-05-28 | 2011-12-01 | Anja Wittke | Milk-based nutritional compositions |
TWI526161B (en) | 2010-06-10 | 2016-03-21 | 亞培公司 | Substantially clear nutritional liquids comprising calcium hmb and soluble protein |
US20120135103A1 (en) | 2010-11-30 | 2012-05-31 | Mead Johnson Nutrition Company | Staged Infant Feeding Regimen To Promote Healthy Development And Growth |
US20120171231A1 (en) | 2010-12-29 | 2012-07-05 | Anja Wittke | Use of nutritional compositions including lactoferrin in stimulating immune cells |
MX2013006094A (en) | 2010-12-29 | 2013-07-03 | Mjn Us Holdings Llc | Method for inhibiting pathogens using a nutritional composition. |
US8968722B2 (en) | 2010-12-29 | 2015-03-03 | Mead Johnson Nutrition Company | Milk-based nutritional compositions containing lactoferrin and uses thereof |
US8648036B2 (en) | 2010-12-29 | 2014-02-11 | Mead Johnson Nutrition Company | Use of nutritional compositions including lactoferrin and one or more prebiotics in inhibiting adhesion of pathogens in the gastrointestinal tract |
CA2823018A1 (en) | 2010-12-29 | 2012-07-05 | Mjn U.S. Holdings Llc | Use of nutritional compositions including lactoferrin in supporting resistance to diseases and conditions |
US20120171328A1 (en) | 2011-01-05 | 2012-07-05 | Dattatreya Banavara | Composition comprising heat labile milk proteins and process for preparing same |
US20120269929A1 (en) | 2011-04-22 | 2012-10-25 | Hugh Lippman | Fortified Milk-Based Nutritional Compositions |
US8365462B2 (en) | 2011-05-31 | 2013-02-05 | Heliae Development, Llc | V-Trough photobioreactor systems |
USD661164S1 (en) | 2011-06-10 | 2012-06-05 | Heliae Development, Llc | Aquaculture vessel |
USD682637S1 (en) | 2011-06-10 | 2013-05-21 | Heliae Development, Llc | Aquaculture vessel |
USD679965S1 (en) | 2011-06-10 | 2013-04-16 | Heliae Development, Llc | Aquaculture vessel |
US20130089638A1 (en) | 2011-10-11 | 2013-04-11 | Mead Johnson Nutrition Company | Compositions Comprising Maltotriose And Methods Of Using Same To Inhibit Damage Caused By Dehydration Processes |
US9474298B2 (en) | 2011-10-11 | 2016-10-25 | Mead Johnson Nutrition Company | Partially hydrolyzed casein-whey nutritional compositions for reducing the onset of allergies |
US20130095204A1 (en) | 2011-10-14 | 2013-04-18 | Zeina Jouni | Nutritional phytonutrient compositions |
US20130095189A1 (en) | 2011-10-14 | 2013-04-18 | Zeina Jouni | Composition and method of phytonutrients for metabolic programming effects |
US9200236B2 (en) | 2011-11-17 | 2015-12-01 | Heliae Development, Llc | Omega 7 rich compositions and methods of isolating omega 7 fatty acids |
US9351978B2 (en) | 2012-02-29 | 2016-05-31 | Mead Johnson Nutrition Company | Neurogenesis screening method and uses thereof |
US20130251829A1 (en) | 2012-03-23 | 2013-09-26 | Mead Johnson Nutrition Company | Probiotic derived non-viable material for infection prevention and treatment |
US20140037813A1 (en) | 2012-08-02 | 2014-02-06 | Mead Johnson Nutrition Company | Nutritional creamer composition |
US20140057014A1 (en) | 2012-08-27 | 2014-02-27 | Carol Lynn Berseth | Formula Fortifier |
US20140170259A1 (en) | 2012-12-14 | 2014-06-19 | Mead Johnson Nutrition Company | Nutritional composition for promoting satiety |
US20140242216A1 (en) | 2013-02-24 | 2014-08-28 | Mead Johnson Nutrition Company | Amino Acid And Protein Hydrolysate Based Formulas With A Stable Emulsion System |
US9873880B2 (en) | 2013-03-13 | 2018-01-23 | Dsm Nutritional Products Ag | Engineering microorganisms |
US9352020B2 (en) | 2013-03-15 | 2016-05-31 | Mead Johnson Nutrition Company | Reducing proinflammatory response |
US9345741B2 (en) | 2013-03-15 | 2016-05-24 | Mead Johnson Nutrition Company | Nutritional composition containing a peptide component with adiponectin simulating properties and uses thereof |
US20140271978A1 (en) | 2013-03-15 | 2014-09-18 | Mead Johnson Nutrition Company | Low-buffer nutritional compositions and uses thereof |
US9345727B2 (en) | 2013-03-15 | 2016-05-24 | Mead Johnson Nutrition Company | Nutritional compositions containing a peptide component and uses thereof |
US9289461B2 (en) | 2013-03-15 | 2016-03-22 | Mead Johnson Nutrition Company | Reducing the risk of autoimmune disease |
US10251928B2 (en) | 2014-11-06 | 2019-04-09 | Mead Johnson Nutrition Company | Nutritional supplements containing a peptide component and uses thereof |
US20140271979A1 (en) | 2013-03-15 | 2014-09-18 | Mead Johnson Nutrition Company | Anti-regurgitation nutritional composition |
US8728546B1 (en) | 2013-03-15 | 2014-05-20 | Swing Aerobics Licensing, Inc. | Medicament for treatment of cancer, cardiovascular diseases and inflammation |
US8889633B2 (en) | 2013-03-15 | 2014-11-18 | Mead Johnson Nutrition Company | Nutritional compositions containing a peptide component with anti-inflammatory properties and uses thereof |
US9138455B2 (en) * | 2013-03-15 | 2015-09-22 | Mead Johnson Nutrition Company | Activating adiponectin by casein hydrolysate |
US20140378419A1 (en) * | 2013-06-21 | 2014-12-25 | Mead Johnson Nutrition Company | Compositions and Methods for Nutrient Delivery |
US9226914B2 (en) | 2013-07-16 | 2016-01-05 | Mead Johnson Nutrition Company | Methods for promoting neuronal development and/or health |
US9241923B2 (en) | 2013-07-16 | 2016-01-26 | Mead Johnson Nutrition Company | Methods for promoting neuronal development and/or health |
US10709770B2 (en) | 2013-07-31 | 2020-07-14 | Mead Johnson Nutrition Company | Nutritional compositions containing a prebiotic and lactoferrin and uses thereof |
US9609888B2 (en) | 2013-07-31 | 2017-04-04 | Mead Johnson Nutrition Company | Nutritional compositions containing synergistic combination and uses thereof |
WO2015050744A2 (en) | 2013-10-03 | 2015-04-09 | Mjn U.S. Holdings Llc | Neurogenesis screening method and uses thereof |
US20150157048A1 (en) | 2013-12-11 | 2015-06-11 | Mead Johnson Nutrition Company | Nutritional compositions containing stearidonic acid and uses thereof |
US20150164833A1 (en) | 2013-12-17 | 2015-06-18 | Mead Johnson Nutrition Company | Nutritional composition containing a neurologic component of ursolic acid and uses thereof |
WO2015094532A1 (en) | 2013-12-17 | 2015-06-25 | Mjn U.S. Holdings Llc | Nutritional composition containing a neurologic component of kaempferol and/or fisetin |
CN115141859A (en) | 2013-12-20 | 2022-10-04 | 玛拉可再生能源公司 | Method for recovering oil from microorganisms |
US10639334B2 (en) | 2014-01-07 | 2020-05-05 | Mead Johnson Nutrition Company | Pediatric nutritional composition with milk peptides for healthy growth and development |
US20150305359A1 (en) | 2014-04-24 | 2015-10-29 | Mead Johnson Nutrition Company | Nutritional compositions directed to subjects having cow's milk protein allergies |
US20150305385A1 (en) | 2014-04-25 | 2015-10-29 | Mead Johnson Nutrition Company | Pediatric nutritional composition with human milk oligosaccahrides, prebiotics and probiotics |
US20160029682A1 (en) | 2014-08-01 | 2016-02-04 | Mead Johnson Nutrition Company | Hydrolyzed lactose-containing nutritional compositions and uses thereof |
SG11201702036QA (en) | 2014-11-07 | 2017-04-27 | Mjn Us Holdings Llc | Nutritional compositions containing a prebiotic and lactoferrin and uses thereof |
AR104042A1 (en) | 2015-03-26 | 2017-06-21 | Mara Renewables Corp | HIGH-DENSITY PRODUCTION OF BIOMASS AND OIL USING GLUCEROL IN GROSS |
US10525016B2 (en) | 2015-06-03 | 2020-01-07 | Mead Johnson Nutrition Company | Nutritional compositions containing an elevated level of inositol and uses thereof |
US10582714B2 (en) | 2015-07-10 | 2020-03-10 | Mead Johnson Nutrition Company | Nutritional compositions and methods for promoting cognitive development |
US10617701B2 (en) | 2015-07-10 | 2020-04-14 | Mead Johnson Nutrition Company | Nutritional compositions containing phosphatidylethanolamine, sphingomyelin and docosahexaenoic acid |
US20170006897A1 (en) | 2015-07-10 | 2017-01-12 | Mead Johnson Nutrition Company | Nutritional compositions and methods for promoting cognitive development |
JP6977231B2 (en) | 2015-07-13 | 2021-12-08 | マラ リニューアブルズ コーポレーション | Enhancement of metabolism of C5 organic carbon by microorganisms |
US20170020950A1 (en) | 2015-07-23 | 2017-01-26 | Mead Johnson Nutrition Company | Methods for modulating kinases |
US9907323B2 (en) | 2015-09-25 | 2018-03-06 | Mead Johnson Nutrition Co. | Infant formula tablets |
US9730969B2 (en) | 2015-11-06 | 2017-08-15 | Mead Johnson Nutrition Company | Nutritional compositions for promoting gut barrier function and ameliorating visceral pain |
US10851395B2 (en) | 2016-06-10 | 2020-12-01 | MARA Renewables Corporation | Method of making lipids with improved cold flow properties |
US20180064739A1 (en) | 2016-09-06 | 2018-03-08 | Mead Johnson Nutrition Company | Nutritional composition with human milk oligosaccharides and uses thereof |
US20180103675A1 (en) | 2016-10-14 | 2018-04-19 | Mead Johnson Nutrition Company | Personalized pediatric nutrition products comprising human milk oligosaccharides |
US20180133287A1 (en) | 2016-11-14 | 2018-05-17 | Mead Johnson Nutrition Company | Nutritional compositions providing dietary management of colic |
US20180153951A1 (en) | 2016-12-05 | 2018-06-07 | Mead Johnson Nutrition Company | Methods for Inducing Adipocyte Browning, Improving Metabolic Flexibility, and Reducing Detrimental White Adipocyte Tissue Deposition and Dysfunction |
US10980269B2 (en) | 2016-12-12 | 2021-04-20 | Mead Johnson Nutrition Company | Protein hydrolysates and methods of making same |
US20180228755A1 (en) * | 2017-02-10 | 2018-08-16 | Taiwan Indigena Botanica Co., Ltd. | Vegetarian composition containing unsaturated fatty acids |
KR102145032B1 (en) * | 2017-04-26 | 2020-08-14 | 주식회사 엘지생활건강 | Cosmetic composition comprising mortierella oil |
US20180333426A1 (en) | 2017-05-17 | 2018-11-22 | Mead Johnson Nutrition Company | Nutritional composition with human milk oligosaccharides and uses thereof |
BR112020002448A2 (en) * | 2017-08-07 | 2020-07-28 | Dsm Ip Assets B.V. | process for the production of concentrated polyunsaturated fatty acid oils |
GB2573538B (en) | 2018-05-09 | 2023-01-04 | Mjn Us Holdings Llc | Pediatric nutritional compositions and methods for infants delivered by C-section |
GB2573539A (en) | 2018-05-09 | 2019-11-13 | Mjn Us Holdings Llc | Wellbeing supplement for postpartum maternal nutrition |
EP4033908A1 (en) | 2019-09-24 | 2022-08-03 | Société des Produits Nestlé S.A. | Glycyrrhiza and the prevention of lc-pufa oxidation |
CN115768271A (en) | 2020-06-17 | 2023-03-07 | 雀巢产品有限公司 | Stabilization of LC-PUFAs by side-stream products from green coffee decaffeination |
EP3933016A1 (en) * | 2020-06-30 | 2022-01-05 | Evonik Operations GmbH | Method of isolating lipids from a lipids containing biomass |
KR102243982B1 (en) * | 2020-08-10 | 2021-04-23 | 주식회사 엘지생활건강 | Composition for improving skin troubles comprising mortierella oil as an active ingredient |
WO2024050590A1 (en) * | 2022-09-07 | 2024-03-14 | Nourish Ingredients Pty Ltd | Compositions and methods for producing meat-like aromas |
Family Cites Families (69)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US2611706A (en) * | 1949-12-21 | 1952-09-23 | Wyeth Corp | Fat composition for infants' food |
US2923628A (en) * | 1957-11-04 | 1960-02-02 | Harold L Otto | Synthetic milk |
US3458625A (en) * | 1963-04-22 | 1969-07-29 | Quaker Oats Co | Ruminant feeding |
US3542560A (en) * | 1967-12-06 | 1970-11-24 | American Home Prod | Infant formula with fat composition like human milk |
US3649295A (en) * | 1970-06-01 | 1972-03-14 | American Home Prod | Humanized fat compositions and infant formulas thereof |
US4058594A (en) * | 1974-04-25 | 1977-11-15 | John Williams | Immuno-suppressive agents |
CH621048A5 (en) * | 1977-04-27 | 1981-01-15 | Nestle Sa | |
US4303692A (en) * | 1978-11-22 | 1981-12-01 | Gaull Gerald E | Infant milk formula |
US4282265A (en) * | 1980-01-14 | 1981-08-04 | Bristol-Myers Company | Fat compositions for infant formulas |
ES507187A0 (en) * | 1981-11-16 | 1983-01-01 | Union Ind Y Agro Ganader S A U | PROCEDURE FOR OBTAINING AN ADDITIONAL HUMANIZED MILK OF NUCLEOTIDES FOR CHILD FEEDING. |
ATE24266T1 (en) * | 1982-04-16 | 1987-01-15 | Nestle Sa | LIPID COMPOSITION FOR ORAL, ENTERAL OR PARENTERAL NUTRITION. |
US4513008A (en) * | 1982-07-30 | 1985-04-23 | The Vinoxen Company, Inc. | Virucidal compositions and therapy |
US4670285A (en) | 1982-08-06 | 1987-06-02 | The University Of Toronto Innovations Foundation | Infant formula |
GB8302708D0 (en) * | 1983-02-01 | 1983-03-02 | Efamol Ltd | Pharmaceutical and dietary composition |
GB8317248D0 (en) | 1983-06-24 | 1983-07-27 | Wyeth John & Brother Ltd | Fat compositions |
JPS6034156A (en) * | 1983-08-08 | 1985-02-21 | Hayashibara Biochem Lab Inc | Eicosapentaenoic acid clathrate compound and food and drink containing the same |
FR2553261B1 (en) * | 1983-10-14 | 1986-02-21 | Bio Extraction | ARTIFICIAL MILK FOR THE FEEDING OF NEWBORNS AND YOUNG CHILDREN |
US4526902A (en) * | 1983-10-24 | 1985-07-02 | Century Laboratories, Inc. | Combined fatty acid composition for treatment or prophylaxis of thrombo-embolic conditions |
DE3470277D1 (en) * | 1984-01-11 | 1988-05-11 | Von Mletzko Armin | Dietetic composition |
GB8404463D0 (en) * | 1984-02-21 | 1984-03-28 | Efamol Ltd | Microbiological production of essential fatty acids |
US4752618A (en) * | 1984-07-12 | 1988-06-21 | New England Deaconess Hospital | Method of minimizing efects of infection through diet |
IT1176916B (en) * | 1984-10-10 | 1987-08-18 | Elvira Pistolesi | PHARMACEUTICAL OR DIETETIC COMPOSITION WITH HIGH ANTI-THROMBOTIC AND ANTI-ARTERIOSCLEROTIC ACTIVITY |
US4870011A (en) | 1985-01-22 | 1989-09-26 | Director General Of Agency Of Industrial Science And Technology | Method for obtaining lipids from fungus bodies |
GB8507058D0 (en) * | 1985-03-19 | 1985-04-24 | Efamol Ltd | Pharmaceutical & dietary compositions |
GB2178752B (en) * | 1985-07-12 | 1989-10-11 | Unilever Plc | Substitute milk fat |
US4792418A (en) * | 1985-08-14 | 1988-12-20 | Century Laboratories, Inc. | Method of extraction and purification of polyunsaturated fatty acids from natural sources |
JPH0716424B2 (en) * | 1985-10-01 | 1995-03-01 | ライオン株式会社 | Method for producing arachidonic acid-containing lipid |
GB8524275D0 (en) * | 1985-10-02 | 1985-11-06 | Efamol Ltd | Pharmaceutical & dietary compositions |
GB8601915D0 (en) * | 1986-01-27 | 1986-03-05 | Efamol Ltd | Pharmaceutical compositions |
DE3603000A1 (en) * | 1986-01-31 | 1987-08-06 | Milupa Ag | NEW FATTY MIXTURE OF POLYENIC ACID AND THEIR USE IN THE PRODUCTION OF INFANT FOODS |
US5204250A (en) * | 1986-03-31 | 1993-04-20 | Suntory Limited | Process for production of arachidonic acid |
US4920098A (en) * | 1986-09-17 | 1990-04-24 | Baxter International Inc. | Nutritional support or therapy for individuals at risk or under treatment for atherosclerotic vascular, cardiovascular, and/or thrombotic diseases |
DE3785667T2 (en) * | 1986-09-17 | 1994-07-21 | Clintec Nutrition Co | ADDITIONAL FOOD OR THERAPY FOR PERSONS WITH RISK OR TREATMENT FOR ARTERIOSCLEROTIC VASCULAR, CARDIOVASCULAR AND / OR THROMBOTIC DISEASES. |
SE8604117D0 (en) * | 1986-09-29 | 1986-09-29 | Kabivitrum Ab | COMPOSITION |
JPS6398355A (en) * | 1986-10-16 | 1988-04-28 | Kazumitsu Maruta | Feed for animal |
JPS63133994A (en) * | 1986-11-21 | 1988-06-06 | Lion Corp | Production of gamma-linolenic acid-containing lipid |
JPS63185389A (en) * | 1987-01-27 | 1988-07-30 | Suntory Ltd | Production of highly unsaturated fatty acid by microbial conversion |
JPS6438007A (en) * | 1987-04-28 | 1989-02-08 | Lion Corp | Skin external preparation |
JPS63295527A (en) * | 1987-05-27 | 1988-12-01 | Nippon Oil & Fats Co Ltd | Method for concentrating and separating highly unsaturated fatty acid |
DE3719097C1 (en) * | 1987-06-06 | 1988-06-09 | Fratzer Uwe | Medicament containing eicosapentaenoic acid and docosahexaenoic acid as unsaturated fatty acids as well as vitamin E. |
GB8714772D0 (en) * | 1987-06-24 | 1987-07-29 | Efamol Ltd | Essential fatty acid compositions |
US5198468A (en) * | 1987-06-24 | 1993-03-30 | Efamol Holdings Plc | Essential fatty acid composition |
KR890701751A (en) * | 1987-07-20 | 1989-12-21 | 원본 미기재 | Microbial Products of Omega-3 (N-3) Lipids |
US4843095A (en) * | 1987-08-07 | 1989-06-27 | Century Laboratories, Inc. | Free fatty acids for treatment or propyhlaxis of rheumatoid arthritis arthritis |
CA1263270A (en) * | 1987-08-19 | 1989-11-28 | Bruce J. Holub | Animal feed supplement |
JP2525624B2 (en) * | 1987-09-21 | 1996-08-21 | 雪印乳業株式会社 | Baby milk powder containing polyunsaturated fatty acids |
JPH0196255A (en) * | 1987-10-09 | 1989-04-14 | Masumi Koishi | Flame-retardant electrical insulating composition |
JP2582622B2 (en) * | 1987-10-27 | 1997-02-19 | 日東化学工業株式会社 | Production of polyunsaturated fatty acids by filamentous fungi |
JPH01132371A (en) * | 1987-11-18 | 1989-05-24 | Itochu Seito Kk | Novel microorganism and production of lipid component of high content of gamma-linolenic acid |
JP2746371B2 (en) * | 1987-12-21 | 1998-05-06 | サントリー株式会社 | Bishomo-γ-linolenic acid and method for producing lipid containing the same |
GB8729751D0 (en) * | 1987-12-21 | 1988-02-03 | Norsk Hydro As | Feed additive & feed containing such additive |
JP2697834B2 (en) * | 1988-02-02 | 1998-01-14 | サントリー株式会社 | Artificial milk with polyunsaturated fatty acid component |
JPH01304892A (en) * | 1988-02-03 | 1989-12-08 | Suntory Ltd | Production of highly unsaturated fatty acid enriched fats and oils |
JP2723243B2 (en) * | 1988-02-25 | 1998-03-09 | サントリー株式会社 | Animal feed with polyunsaturated fatty acids |
US4874629A (en) * | 1988-05-02 | 1989-10-17 | Chang Stephen S | Purification of fish oil |
GB8813766D0 (en) * | 1988-06-10 | 1988-07-13 | Efamol Holdings | Essential fatty acid compositions |
US5130242A (en) * | 1988-09-07 | 1992-07-14 | Phycotech, Inc. | Process for the heterotrophic production of microbial products with high concentrations of omega-3 highly unsaturated fatty acids |
US5116871A (en) * | 1988-09-13 | 1992-05-26 | Efamol Holdings Plc | Fatty acid therapy and compositions for the treatment of myalgic encephalomyelitis |
JP2730081B2 (en) * | 1988-09-24 | 1998-03-25 | 日本油脂株式会社 | Method for producing arachidonic acid-containing fats and oils by microorganism |
GB2223943A (en) * | 1988-10-21 | 1990-04-25 | Tillotts Pharma Ag | Oral disage forms of omega-3 polyunsaturated acids |
US5935828A (en) * | 1989-05-01 | 1999-08-10 | Opta Food Ingredients, Inc. | Enzymatic production of monoglycerides containing omega-3 unsaturated fatty acids |
US4963385A (en) * | 1989-06-02 | 1990-10-16 | Nabisco Brands, Inc. | Stabilized emulsions containing highly unsaturated oils |
DE3920679A1 (en) * | 1989-06-23 | 1991-01-10 | Milupa Ag | FAT MIXTURE FOR THE MANUFACTURE OF FOOD, ESPECIALLY SUGAR FOODS |
US5407957A (en) | 1990-02-13 | 1995-04-18 | Martek Corporation | Production of docosahexaenoic acid by dinoflagellates |
US5244921A (en) | 1990-03-21 | 1993-09-14 | Martek Corporation | Eicosapentaenoic acids and methods for their production |
US5013569A (en) * | 1990-05-21 | 1991-05-07 | Century Laboratories, Inc. | Infant formula |
PH11992043811B1 (en) * | 1991-01-24 | 2002-08-22 | Martek Corp | Arachidonic acid and methods for the production and use thereof |
FR2686619B1 (en) * | 1992-01-28 | 1995-07-13 | Commissariat Energie Atomique | PROCESS FOR THE SELECTIVE PRODUCTION OF POLYUNSATURATED LIPIDS FROM A CULTURE OF PORPHYRIDIUM MICROALGAE AND TANK USED IN THIS PROCESS. |
US5234702A (en) * | 1992-03-19 | 1993-08-10 | Abbott Laboratories | Antioxidant system for powdered nutritional products |
-
1992
- 1992-01-22 AU AU12392/92A patent/AU661297B2/en not_active Expired
- 1992-01-22 AT AT92904388T patent/ATE200619T1/en not_active IP Right Cessation
- 1992-01-22 ZA ZA92452A patent/ZA92452B/en unknown
- 1992-01-22 ES ES92904388T patent/ES2157898T3/en not_active Expired - Lifetime
- 1992-01-22 DE DE69231793T patent/DE69231793T2/en not_active Revoked
- 1992-01-22 EP EP92904388A patent/EP0568606B1/en not_active Revoked
- 1992-01-22 DK DK92904388T patent/DK0568606T3/en active
- 1992-01-22 EP EP07075341A patent/EP1832181A3/en not_active Withdrawn
- 1992-01-22 NZ NZ241359A patent/NZ241359A/en not_active IP Right Cessation
- 1992-01-22 EP EP01200067A patent/EP1092352A3/en not_active Withdrawn
- 1992-01-22 JP JP4504606A patent/JP2731035B2/en not_active Ceased
- 1992-01-22 EP EP07002670A patent/EP1787532A3/en not_active Withdrawn
- 1992-01-22 RU RU9293052410A patent/RU2093996C1/en active
- 1992-01-22 WO PCT/US1992/000522 patent/WO1992012711A1/en not_active Application Discontinuation
- 1992-01-22 DE DE0568606T patent/DE568606T1/en active Pending
- 1992-01-22 KR KR1019930702205A patent/KR100321543B1/en active
- 1992-01-22 SG SG1996008986A patent/SG49307A1/en unknown
- 1992-01-22 BR BR9205526A patent/BR9205526A/en not_active Application Discontinuation
- 1992-01-22 IL IL114253A patent/IL114253A/en not_active IP Right Cessation
- 1992-01-22 EP EP08005794A patent/EP1961312A3/en not_active Withdrawn
- 1992-01-22 CA CA002101274A patent/CA2101274C/en not_active Expired - Lifetime
- 1992-01-22 IL IL10073392A patent/IL100733A/en not_active IP Right Cessation
- 1992-01-24 MX MX9200320A patent/MX183638B/en unknown
- 1992-09-14 US US07/944,739 patent/US5374657A/en not_active Expired - Lifetime
-
1993
- 1993-07-23 OA OA60396A patent/OA10348A/en unknown
-
1994
- 1994-12-19 US US08/358,474 patent/US5550156A/en not_active Expired - Lifetime
-
2000
- 2000-03-30 KR KR20007003480A patent/KR100292103B1/en not_active IP Right Cessation
-
2001
- 2001-02-22 KR KR20017002283A patent/KR100313987B1/en not_active IP Right Cessation
- 2001-06-27 GR GR20010400993T patent/GR3036139T3/en not_active IP Right Cessation
Also Published As
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CA2101274C (en) | Microbial oil mixtures and uses thereof | |
CA2209513C (en) | Arachidonic acid and methods for the production and use thereof | |
AU766660B2 (en) | Utilization of material containing docosapentaenoic acid | |
RU2120998C1 (en) | Method of preparing arachidonic-containing oil, nonmodified microbial oil, method of providing a mixture for children nutrition with an arachidonic acid, cosmetic composition, nutrient or nutritious addition, method of human treatment and a mixture for children nutrition | |
JP4761771B2 (en) | Method for producing microbial fats and oils with reduced unsaponifiable matter content and said fats and oils | |
US5711983A (en) | Dinoflagellate biomass, methods for its production, and compositions containing the same | |
JP3985035B2 (en) | (N-6) Docosapentaenoic Acid-Containing Oil and Fat, Method for Producing the Oil and Use, and Use | |
IL164123A (en) | Aquatic animal feed containing microalgae containing arachidonic acid | |
WO1996038051A1 (en) | Fowl eggs with high content of highly unsaturated fatty acids, process for producing the same, and use thereof | |
WO2023145785A1 (en) | Thrombosis preventive agent | |
Mokady et al. | A marine unicellular alga in diets of pregnant and lactating rats as a source of ω3 fatty acids for the developing brain of their progeny | |
KR20010010268A (en) | Arachidonic Acid Reinforced Feed Composition |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
EEER | Examination request | ||
MKEX | Expiry | ||
MKEX | Expiry |
Effective date: 20120122 |