Process for preparing materials for extraction

• US 7,678,931 B2
• Filed: 10/21/2005
• Issued: 03/16/2010
• Est. Priority Date: 10/22/2004
• Status: Expired due to Fees

• Current Assignee: Martek Biosciences Corporation (DSM-Firmenich AG)
• Original Assignee: Martek Biosciences Corporation (DSM-Firmenich AG)
• Inventors: Sundararajan, Anand, Fichtali, Jaouad
• Primary Examiner(s): Saucier; Sandra
• Assistant Examiner(s): Singh; Satyendra K
• Application Number:
  US11/255,787
• Publication Number:
  US 20060122410A1
• Time in Patent Office: 1,607 Days
• Field of Search: None
• US Class Current: 554/20
• CPC Class Codes: A23L 33/12 Fatty acids or derivatives thereof
• CPC Class Codes: A61K 36/00 Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines antigens from pollen A61K39/36
• CPC Class Codes: A61K 36/062 Ascomycota
• CPC Class Codes: C11B 1/02 Pretreatment
• CPC Class Codes: C11B 1/10 by extracting
• CPC Class Codes: C12N 1/005 after treatment of microbial biomass not covered by C12N1/02 - C12N1/08
• 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

Abstract

The present invention relates to a process for preparing a biomass, such as from a microbial fermentation, for an extraction process to separate desired chemicals, nutritional products, bioactive components, proteins, carbohydrates, and lipids, from the biomass. Particularly preferred substances to extract include docosahexaenoic acid, docosapentaenoic acid, and arachidonic acid. The present invention also includes extracting the prepared biomass. Biomasses to be treated in accordance with the methods of the invention include plant, animal, and microbial biomass, particularly a microorganism such as Crypthecodinium cohnii and a fungus such as Mortierella alpina.

Claims

1. A method for extracting a lipid from a microbial biomass, comprising:

(a) preparing the microbial biomass by a process, comprising;

(i) freezing the microbial biomass at or below its brittleness temperature to produce a frozen microbial biomass;

(ii) comminuting the frozen microbial biomass at a temperature no higher than the brittleness temperature to produce a comminuted biomass;

(iii) thawing the comminuted biomass to at least a temperature at which the comminuted biomass is deformable;

(iv) adding an effective amount of a dry agent to the thawed comminuted biomass to achieve an extrudable consistency;

(v) extruding the comminuted biomass from step (iv) to produce an extruded biomass; and

(b) percolation extracting the lipid from the extruded biomass.

2. The method of claim 1, wherein at least 70% by weight of the lipids in the biomass are extracted.

3. The method of claim 1, wherein at least 90% by weight of the lipids in the biomass are extracted.

4. The method of claim 1, further comprising pasteurizing the biomass.

5. The method of claim 4, wherein pasteurizing is performed prior to the freezing.

6. The method of claim 1, further comprising separating the biomass, prior to step (a), from a culture medium in which the biomass was cultured.

7. The method of claim 6, wherein the separated biomass is dried.

8. The method of claim 7, wherein the drying is selected from the group consisting of centrifugation, filtration, membrane filter press drying, spray drying, fluidized bed drying, freeze drying, tray drying, vacuum tray drying, drum drying, and vacuum mixer/reactor drying.

9. The method of claim 7, wherein the moisture content of the biomass is less than about 25% by weight.

10. The method of claim 1, wherein an antioxidant is added to the biomass.

11. The method of claim 10, wherein the antioxidant is added to the biomass prior to freezing.

12. The method of claim 10, wherein the antioxidant is selected from the group consisting of ascorbyl palmitate, tocopherols, citric acid, ascorbic acid, tertiary butyl hydroquinone (TBHQ), rosemary extract, lecithin, and mixtures thereof.

13. The method of claim 1, wherein the biomass comprises a microorganism selected from the group consisting of an algae, bacteria, fungi and protist.

14. The method of claim 13, wherein the microorganism is a marine microorganism selected from the group consisting of order Dinophyceae (Dinoflagellates), Stramenopiles (golden algae), and order Thraustochytriales.

15. The method of claim 13, wherein the microorganism is a microorganism selected from the group consisting the genus Thraustochytrium, genus Schizochytrium, genus Althornia, genus Aplanochytrium, genus Japonochytrium, genus Labyrinthula, genus Labyrithuloides, genus Crypthecodinium, and genus Mortierella.

16. The method of claim 13, wherein the microorganism is selected from the group consisting of Crypthecodinium cohnii and the fungus Mortierella alpina.

17. The method of claim 1, where freezing is accomplished by a method selected from the group consisting of air freezing, blast freezing, fluidized-bed freezing, plate freezing, liquid immersion freezing, cryogenic freezing, liquid nitrogen freezing, dry ice freezing, and CCl₂F₂ freezing.

18. The method of claim 1, wherein comminuting is accomplished by the use of a comminution device selected from the group consisting of a crushing device, a grinding device, and a homogenization device.

19. The method of claim 1, wherein comminuting is accomplished by a pin mill.

20. The method of claim 1, wherein comminuting results in the comminuted biomass having a particle size range from about 8 μ

m to about 50 μ

m.

21. The method of claim 1, wherein extruding takes place at a temperature between about 10°

C. and about 60°

C.

22. The method of claim 1, wherein extruding takes place at a temperature of from about 20°

C. and about 30°

C.

23. The method of claim 1 wherein the dry agent is selected from the group consisting of plant starches, plant fibers, biomeal, oilseed hulls, rice flour and peanut hulls.

24. The method of claim 1 wherein the dry agent is added to the biomass at a final concentration of between about 1% by dry weight and about 50% by dry weight.

25. The method of claim 1 wherein the dry agent is added to the biomass at a final concentration of between about 5% by dry weight and about 30% by dry weight.

26. The method of claim 1 wherein the moisture content of the comminuted biomass prior to extrusion is less than about 25% by weight.

27. The method of claim 1, wherein the moisture content of the comminuted biomass prior to extrusion is about 10% by weight.

28. The method of claim 1, wherein about 50% of the volume of the comminuted biomass comprises air.

29. The method of claim 1, wherein the lipid is selected from the group consisting of acylglycerols, phosphoglycerides, fatty acids, sphingolipids, gangliosides, phospholipids, waxes, tocopherols, tocotrienols, triacylglycerols, sterols, carotenoids, pigments, polyphenols, and antioxidants.

30. The method of claim 1, wherein the lipid comprises a PUFA having a carbon chain length of at least 20.

31. The method of claim 1, wherein the lipid comprises a PUFA having a carbon chain length of at least 22.

32. The method of claim 1, wherein the lipid comprises a PUFA having at least three double bonds.

33. The method of claim 1, wherein the lipid comprises a PUFA comprising a lipid having at least four double bonds.

34. The method of claim 1, wherein the lipid comprises docosahexaenoic acid.

35. The method of claim 34, wherein the lipid comprises at least 55 weight percent docosahexaenoic acid.

36. The method of claim 1, wherein the lipid comprises docosapentaenoic acid.

37. The method of claim 1, wherein the lipid comprises arachidonic acid.

38. The method of claim 1, wherein the lipid comprises eicosapentaenoic acid.

39. The method of claim 1, wherein the percolation extraction is selected from the group consisting of aqueous solvent extraction, organic solvent extraction, near-critical solvent extraction, supercritical solvent extraction, enzyme-assisted extraction, microwave extraction and mechanical extraction.

40. The method of claim 1, wherein the percolation extraction is organic solvent extraction with a solvent selected from the group consisting of hexane and isohexane.

41. The method of claim 1, further comprising drying the extruded biomass prior to percolation extraction.