Process for preparing materials for extraction

• US 20060122410A1
• Filed: 10/21/2005
• Published: 06/08/2006
• Est. Priority Date: 10/22/2004
• Status: Active Grant

• Current Assignee: Martek Biosciences Corporation (DSM-Firmenich AG)
• Original Assignee: Martek Biosciences Corporation (DSM-Firmenich AG)
• Inventors: Fichtali, Jaouad, Sundararajan, Anand
• Granted Patent: US 7,678,931 B2
• Time in Patent Office: Days
• US Class Current: 554/8
• 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 preparing a biomass for extraction comprising:

(a) freezing the biomass at or below its brittleness temperature;

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

(c) agglomerating the comminuted biomass to produce an agglomerated biomass.

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

3. The method of claim 2, wherein the pasteurizing step is performed prior to the freezing step.

4. The method of claim 1, further comprising separating the biomass from a culture media in which the biomass was cultured.

5. The method of claim 4, wherein the separated biomass is dried.

6. The method of claim 5, wherein the drying step is selected from the group consisting of centrifugation, filtration, membrane filter press drying, spray drying, fluidized bed drying, lyophilization, freeze drying, tray drying, vacuum tray drying, drum drying, and vacuum mixer/reactor drying.

7. The method of claim 5, wherein the drying step is spray drying.

8. The method of claim 4, wherein the moisture content of the biomass is less than about 25% by weight.

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

10. The method of claim 9, wherein the antioxidant is added to the biomass prior to the freezing step.

11. The method of claim 9, 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.

12. The method of claim 1, wherein the biomass comprises a biological material selected from the group consisting of an animal material, a plant material, and a microorganism.

13. The method of claim 12, 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 the freezing step 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 the freezing step is accomplished by the step of liquid nitrogen freezing

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

20. The method of claim 1, wherein the comminuting step is accomplished by a pin mill.

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

m to about 50 μ

m.

22. The method of claim 1, wherein the step of agglomerating comprises extruding the biomass particles.

23. The method of claim 22, wherein the extruding step takes place at a temperature between about 10 C and about 60 C.

24. The method of claim 22, wherein the extruding step takes place at a temperature of from about 20 C to about 30 C.

25. The method of claim 1, wherein the biomass is mixed with a dry agent prior to or during the agglomerating step.

26. The method of claim 25 wherein the dry agent is selected from the group consisting of plant starches, plant fibers, biomeal, and oilseed hulls.

27. The method of claim 25 wherein the dry agent is selected from the group consisting of rice flour and peanut hulls.

28. The method of claim 25 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.

29. The method of claim 25 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.

30. The method of claim 25 wherein the moisture content of the agglomerated biomass is less than about 25% by weight.

31. The method of claim 25, wherein the moisture content of the agglomerated biomass is about 10% by weight.

32. The method of claim 25, wherein about 50% of the volume of the agglomerated particle comprises air.

33. A method of extracting a substance from a biomass, comprising:

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

(i) freezing the biomass at or below its brittleness temperature;

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

(iii) agglomerating the biomass to produce an agglomerated biomass; and

(b) extracting the desired substance from the agglomerated particles.

34. The method of claim 33, wherein the desired substance comprises a lipid.

35. The method of claim 34, wherein the lipid is selected from the group consisting of acylglycerols, phosphoglycerides, fatty acids, sphingolipids, gangliosides, phospholipids, waxes, tocopherols, tocotrienols, sterols, carotenoids, pigments, polyphenols, and antioxidants.

36. The method of claim 34, wherein the lipid is selected from the group consisting of triacylglycerols and phospholipids.

37. The method of claim 34, wherein the lipid comprises a PUFA having a carbon chain length of at least 20.

38. The method of claim 34, wherein the lipid comprises a PUFA having a carbon chain length of at least 22.

39. The method of claim 34, wherein the lipid comprises a PUFA having at least three double bonds.

40. The method of claim 34, wherein the lipid comprises a PUFA comprising a lipid having at least four double bonds.

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

42. The method of claim 41, wherein the lipid comprises at least about 55 weight percent docosahexaenoic acid.

43. The method of claim 34, wherein the lipid comprises docosapentaenoic acid.

44. The method of claim 34, wherein the lipid comprises arachidonic acid.

45. The method of claim 34, wherein the lipid comprises eicosapentaenoic acid.

46. The method of claim 34, wherein the desired substance comprises a protein.

47. The method of claim 33, wherein the desired substance comprises a substance selected from the group consisting of a saccharide, an isoflavone, a flavonoid, a phytochemical, an antioxidant, a lutein, a lycopene, and a saponin.

48. The method of claim 33, wherein the extraction step 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.

49. The method of claim 48, wherein the extraction step is organic solvent extraction with a solvent selected from the group consisting of hexane and isohexane.

50. The method of claim 33, wherein the extraction step is performed with an extraction device selected from the group consisting of a percolation-type extractor and an immersion-type extractor.

51. The method of claim 33, wherein the extraction step is performed with a percolation-type extractor.

52. The method of claim 33, further comprising drying the agglomerated biomass prior to the extraction step.

53. The method of claim 52, wherein the drying step comprises spray drying.

54. 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;

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

(iii) adding a dry agent to the comminuted biomass;

(iv) extruding the biomass to produce an extruded biomass; and

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

55. The method of claim 54, wherein at least about 70% by weight of the lipids in the biomass are extracted.

56. The method of claim 54, wherein at least about 90% by weight of the lipids in the biomass are extracted.

57. A biomass useful for extraction produced by a process, comprising:

(a) freezing the biomass at or below its brittleness temperature;

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

(c) agglomerating the comminuted biomass to produce an agglomerated biomass.

58-70. -70. (canceled)

71. A biomass useful for extraction produced by a proces, comprising:

(a) freezing a biomass comprising a marine microorganism selected from the group consisting of order Dinophyceae (Dinoflagellates), Stramenopiles (golden algae), and order Thraustochytriales at or below its brittleness temperature;

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

(c) agglomerating the comminuted biomass to produce an agglomerated biomass.

72-80. -80. (canceled)