Process for the Heterotrophic Production of Microbial Products with High Concentrations of Omega-3 Highly Unsaturated Fatty Acids

• US 20080199923A1
• Filed: 10/30/2007
• Published: 08/21/2008
• Est. Priority Date: 10/16/1992
• Status: Active Grant

• Current Assignee: DSM IP Assets BV (DSM-Firmenich AG)
• Original Assignee: Martek Biosciences Corporation (DSM-Firmenich AG)
• Inventors: Barclay, William R.
• Granted Patent: US 8,288,135 B2
• Time in Patent Office: Days
• US Class Current: 435/134
• CPC Class Codes: A23K 10/16 Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions A23K10/12 takes precedence
• CPC Class Codes: A23K 20/158 Fatty acids; Fats; Products containing oils or fats
• CPC Class Codes: A23K 40/20 by moulding, e.g. making cakes or briquettes
• CPC Class Codes: A23K 40/25 by extrusion
• CPC Class Codes: A23K 50/00 Feeding-stuffs specially adapted for particular animals
• CPC Class Codes: A23K 50/10 for ruminants
• CPC Class Codes: A23K 50/75 for poultry
• CPC Class Codes: A23K 50/80 for aquatic animals, e.g. fish, crustaceans or molluscs
• CPC Class Codes: A23L 11/07 Soya beans, e.g. oil-extracted soya bean flakes A23L11/30 takes precedence
• CPC Class Codes: A23L 13/00 Meat products; Meat meal; Preparation or treatment thereof
• CPC Class Codes: A23L 13/43 Addition of vegetable fats or oils; Addition of non-meat animal fats or oils; Addition of fatty acids
• CPC Class Codes: A23L 13/50 Poultry products, e.g. poultry sausages
• CPC Class Codes: A23L 15/20 Addition of proteins, e.g. hydrolysates, fats, carbohydrates, natural plant hydrocolloids; Addition of animal or vegetable substances containing proteins, fats, or carbohydrates
• CPC Class Codes: A23L 17/00 Food-from-the-sea products; Fish products; Fish meal; Fish-egg substitutes; Preparation or treatment thereof
• CPC Class Codes: A23L 17/40 Shell-fish
• CPC Class Codes: A23L 17/65 Addition of, or treatment with, microorganisms or enzymes
• CPC Class Codes: A23L 19/09 Mashed or comminuted products, e.g. pulp, purée, sauce, or products made therefrom, e.g. snacks A23L19/01, A23L19/10, A23L27/63 and A23L21/00 take precedence
• CPC Class Codes: A23L 25/30 Mashed or comminuted products, e.g. pulp, pastes, meal, powders; Products made therefrom, e.g. blocks, flakes, snacks; Liquid or semi-liquid products A23L25/10 and A23L25/40 take precedence
• CPC Class Codes: A23L 29/065 Microorganisms addition of bacteria for nutritional purposes A23L33/135
• CPC Class Codes: A23L 33/12 Fatty acids or derivatives thereof
• CPC Class Codes: A61K 31/20 : having a carboxyl group bound to a chain of seven or more carbon atoms, e.g. stearic, palmitic, arachidic acids
• CPC Class Codes: A61K 31/202 : having three or more double bonds, e.g. linolenic eicosanoids, e.g. leukotrienes A61K31/557
• CPC Class Codes: C11B 1/10 : by extracting
• CPC Class Codes: C12N 1/10 : Protozoa; Culture media therefor
• CPC Class Codes: C12N 1/12 : Unicellular algae; Culture media therefor as new plants A01H13/00
• CPC Class Codes: C12N 1/14 : Fungi culture of mushrooms A01G18/00; as new plants A01H15/00; Culture media therefor
• CPC Class Codes: C12P 7/6409 : Fatty acids
• CPC Class Codes: C12P 7/6427 : Polyunsaturated fatty acids [PUFA], i.e. having two or more double bonds in their backbone
• CPC Class Codes: C12P 7/6432 : Eicosapentaenoic acids [EPA]
• CPC Class Codes: C12P 7/6463 : obtained from glyceride producing microorganisms, e.g. single cell oil
• CPC Class Codes: C12P 7/6472 : containing polyunsaturated fatty acid [PUFA] residues, i.e. having two or more double bonds in their backbone
• CPC Class Codes: C12P 7/649 : Biodiesel, i.e. fatty acid alkyl esters
• CPC Class Codes: Y02A 40/818 : Alternative feeds for fish, e.g. in aquacultures
• CPC Class Codes: Y02E 50/10 : Biofuels, e.g. bio-diesel
• CPC Class Codes: Y10S 426/805 : Pet food for dog, cat, bird, or fish
• CPC Class Codes: Y10S 426/807 : Poultry or ruminant feed
• CPC Class Codes: Y10S 435/911 : using fungi
• CPC Class Codes: Y10S 435/946 : using algae

Abstract

Disclosed is a process for growing the microflora Thraustochytrium, Schizochytrium, and mixtures thereof, which includes the growing of the microflora in fermentation medium containing non-chloride containing sodium salts, in particular sodium sulfate. In a preferred embodiment of the present invention, the process produces microflora having a cell aggregate size useful for the production of food products for use in aquaculture. Further disclosed is a food product which includes Thraustochytrium, Schizochytrium, and mixtures thereof, and a component selected from flaxseed, rapeseed, soybean and avocado meal. Such a food product includes a balance of long chain and short chain omega-3 highly unsaturated fatty acids.

Claims

1-73. -73. (canceled)

74. A process for culturing a microorganism of the order Thraustochytriales for the production of lipids, the process comprising:

culturing the microorganism of the order Thraustochytriales in a culture medium that comprises a chloride concentration of less than about 3 grams of chloride per liter of said culture medium and that comprises a non-chloride source of sodium selected from the group consisting of soda ash, sodium carbonate, sodium bicarbonate, sodium sulfate and mixtures thereof, and wherein the microorganism has a sterol content of at least about 0.1% ash-free dry weight (afdw) and a cholesterol content of at least about 15% of the total sterol content.

75. The process of claim 74, wherein the non-chloride source of sodium provides a sodium concentration in a range of from about 1 g/L to about 50 g/L.

76. The process of claim 74, wherein the culture medium comprises between about 60 mg and about 120 mg of chloride per liter of the culture medium.

77. The process of claim 74, wherein the non-chloride source of sodium provides a sodium concentration in a range of from about 2 g/L to about 25 g/L.

78. The process of claim 74, wherein the lipids comprise at least one highly unsaturated fatty acid (HUFA).

79. The process of claim 78, wherein the at least one HUFA is selected from the group consisting of DHA, EPA, DPA(n-3), and mixtures thereof.

80. The process of claim 79, wherein the at least one HUFA comprises DHA.

81. The process of claim 74, wherein the non-chloride source of sodium provides a sodium concentration in an amount less than about 25 g/L.

82. The process of claim 74, wherein a source of chloride comprises sodium chloride.

83. The process of claim 74, wherein the microorganism produces up to about 12% total cellular dry weight percent (% dwt) omega-3 fatty acids.

84. The process of claim 74, wherein the microorganism replicates by successive bipartition.

85. The process of claim 74, wherein the microorganism replicates by forming sporangia and releasing zoospores.

86. The process of claim 74, wherein the microorganism is Thraustochytrium, Schizochytrium or mixtures thereof.

87. The process of claim 74, further comprising culturing the microorganism at a temperature from about 5°

C. to about 48°

C. and a pH from about pH 5.0 to about pH 11.0.

88. The process of claim 74, wherein a non-chloride source of sodium is sodium sulfate.

89. The process of claim 74, wherein less than about 25% of the sodium in the culture medium is supplied as sodium chloride.

90. The process of claim 74, wherein the microorganism has an average cell aggregate size less than about 150 microns in diameter.

91. The process of claim 74, wherein the microorganism has an average cell aggregate size less than about 100 microns in diameter.

92. The process of claim 74, wherein the microorganism has an average cell aggregate size less than about 50 microns in diameter.

93. The process of claim 74, wherein the culturing produces a yield of biomass per sugar of about 50% or greater.

94. The process of claim 74, further comprising the step of recovering fatty acids from the microorganism.