Fermentation process

• US 20040115779A1
• Filed: 09/19/2003
• Published: 06/17/2004
• Est. Priority Date: 03/19/2002
• Status: Abandoned Application

• Current Assignee: Novozymes A/S (Novozymes), Novozymes North America Incorporated (Novozymes)
• Original Assignee: Novozymes A/S (Novozymes), Novozymes North America Incorporated (Novozymes)
• Inventors: Felby, Claus, Beckerich, Robert, Pedersen, Sven, Olsen, Hans Sejr, Veit, Christopher
• Application Number:
  US10/472,256
• Publication Number:
  US 20040115779A1
• Time in Patent Office: Days
• US Class Current: 435/105
• CPC Class Codes: C12C 5/004 Enzymes
• CPC Class Codes: C12P 19/20 produced by the action of an exo-1,4 alpha-glucosidase, e.g. dextrose
• CPC Class Codes: C12P 7/06 Ethanol, i.e. non-beverage
• CPC Class Codes: Y02E 50/10 Biofuels, e.g. bio-diesel

Abstract

The present invention relates to an improved process for producing a fermentation product.

Claims

1. A process of producing a fermentation product, which process comprises a fermentation step, wherein the fermentation step is carried out in the presence of at least one carbohyrate-source generating-enzyme activity and at least one alpha-amylase activity.

2. The process of claim 1, wherein further a protease is present.

3. The process of claim 1 or 2, wherein further a debranching enzyme activity is present.

4. The process of claims 1-3, wherein the carbohydrate-source generating enzyme is a glucoamylase, in particular derived from Aspergillus niger or Talaromyces emersonli;

or beta-amylase, in particular derived from barley;

or a maltogenic amylase, in particular derived from Bacillus stearothermophilus.

5. The process of claims 1-4, wherein the alpha-amylase is an acid alpha-amylase, in particular an acid fungal alpha-amylase, such as an acid fungal alpha-amylase derived from Aspergillus niger or Aspergillus oryzae.

6. The process of claims 1-5, wherein the ratio between acid fungal alpha-amylase activity (AFAU) per glucoamylase activity (AGU) (AFAU per AGU) is at least 0.1, in particular at least 0.16, especially in the range from 0.12 to 0.30.

7. The process of claims 2-6, wherein the protease is an acid protease, in particular an acid fungal protease, such as an acid fungal protease derived from a strain of Aspergillus, in particular Aspergillus niger or Aspergillus oryzae;

or a strain of Rhizomucor, in particular Rhizomucor miehei.

8. The process of claim 3-7, wherein the debranching enzyme is an isoamylase (E.C. 3.2.1.68) or pullulanase (E.C. 3.2.1.41), in particular a pullulanase derived from Bacillus sp., such as a strain of Bacillus deramificans;

Bacillus acidopullulyticus or Bacillus naganoensis.

9. The process of claims 3-8, wherein the glucoamylase/pullulanase ratio determined as AGU/PUN is from 5:

1 to 1;

5.

10. The process of claims 1-9, wherein the fermentation is carried out using a micro-organism capable of fermenting sugars or converted sugars, such as glucose or maltose.

11. The process of claims 1-10, wherein the micro-organism is a yeast, in particular Saccharomyces cerevisae.

12. The process of claims 1-11, wherein the material to be fermented is liquefied whole grain mash or a side stream from starch processing, in particular liquefied starch with a DE of 8-10.

13. The process of claims 1-12, wherein the product is beer or wine

14. The process of claims 1-12, wherein the product is ethanol.

26. The process of any of claims 1-25 wherein further one or more yeast cell wall degrading enzyme(s) is(are) present.

60. The process of any of claims 1-26 and 34-59 where one or more process step(s) is(are) performed batch wise and/or one or more process step(s) is(are performed as a continuous flow

15. A process for the production of ethanol, comprising the steps of:

(a) milling whole grains;

(b) liquefying the product of step (a), in the presence of an alpha-amylase;

(c) saccharifying the liquefied material obtained in step (b);

(d) fermenting the saccharified material obtained in step (c) using a micro-organism, and optionally;

(e) distilling of the fermented and saccharified material obtained in step (d) providing two fraction;

     1) an alcohol fraction and

     2) a Whole Stillage fraction;

(f) separating the Whole Stillage into two fractions;

     1) Wet Grain fraction, and

     2) Thin Stillage;

(g) optionally the Thin Stillage is evaporated to provide two fractions;

     1) Condensate and

     2) Syrup;

wherein the steps (c) and/or (d) are carried out in the presence of carbohydrate-source generating enzyme activity and an alpha-amylase activity and the and steps (c) and (d) may be carried out either simultaneously or separately/sequentially.

16. The process of claim 15, wherein the liquefaction step comprises the following sub-steps;

b1) the hot slurry is heated to between 60-95°

C., preferably 80-85°

C., and at least one alpha-amylase is added;

b2) the slurry is jet-cooked at a temperature between 95-140°

C., preferably 105-125°

C. to complete gelatinization of the slurry;

b3) the slurry is cooled to 60-95°

C. and more alpha-amylase is added to finalize hydrolysis.

17. The process of claims 15 or 16, wherein the liquefaction process is carried out at pH 4.5-6.5, in particular at a pH between 5 and 6.

18. The process of claims 15-17, wherein further a protease activity, and/or debranching enzyme activity is added during pre-saccharification or saccharification or SSF.

19. The process of claims 15-18, wherein further a protease activity and/or debranching enzyme activity is added during fermentation.

20. The process of claims 15-19, wherein steps (c) and (d) are carried out either simultaneously or separately/sequentially.

21. The process of claims 15-20, wherein an optionally ethanol recovery step is carried out after step (e).

22. The process of claim 15-21, wherein the milling step (a) is a dry milling step or wet milling step.

23. The process of claims 15-22, wherein the alpha-amylase(s) used for liquefaction in step (b) is derived from a strain of the genus Bacillus or a strain of Aspergillus.

24. The process of claims 15-23, wherein the fermentation is carried out using a micro-organism capable of fermenting sugars to ethanol.

25. The process of claim 24, wherein the micro-organism is a yeast, such as a yeast belonging to Saccharomyces spp., in particular Saccharomyces cerevisae.

27. A composition comprising all of the following enzyme activities:

carbohydrate-source generating enzyme activity, alpha-amylase activity, protease activity and debranching enzyme activity.

28. Use of a composition of claim 27 for saccharification and/or fermentation.

29. Use of a composition of claim 27 for ethanol production or for beer or wine production.

30. The composition of claim 27 wherein further one or more yeast cell wall degrading enzyme(s) is(are) present.

31. Use of a composition of claim 30 for saccharification and/or fermentation.

32. Use of a composition of claim 30 for beer or wine production

33. Use of a composition of claim 30 for ethanol production.

44. A process of producing ethanol, wherein the process sequentially comprises the following steps

a) milling whole grains;

b) liquefying the product of step (a), in the presence of an alpha-amylase;

c) saccharifying the liquefied material obtained in step (b);

d) fermenting the saccharified material obtained in step (c) using a micro-organism, and optionally;

e) distilling the fermented and saccharified material obtained in step (d), providing two fraction;

     1) an alcohol fraction and

     2) a Whole Stillage fraction;

f) separating the Whole Stillage into two fractions;

     1) Wet Grain fraction, and

     2) Thin Stillage;

g) optionally the Thin Stillage is evaporated to provide two fractions;

     1) Condensate and

     2) Syrup;

wherein step (c ) and/or (d) is carried out in the presence of one or more yeast cell wall degrading enzyme(s), in particular one more enzyme activities as defined in claims 33 or 34.

45. The process of claim 44, wherein the treated Thin Stillage and/or condensate thereof is recycled to the liquefaction step b), in particular to the milled whole grain slurry.

46. The process of claims 44 or 45, wherein the liquefaction step comprises the following sub-steps:

b1) the hot slurry is heated to between 60-95°

C., preferably 80-85°

C., and at least one alpha-amylase is added;

b2) the slurry is jet-cooked at a temperature between 95-140°

C., preferably 105-125°

C. to complete gelanitization of the slurry;

b3) the slurry is cooled to 60-95°

C. and more alpha-amylase is added to finalize hydrolysis.

47. The process of claims 44-46, wherein the liquefaction process is carried out at pH 4.5-6.5, in particular at a pH between 5 and 6.

48. The process of claims 44-47, wherein the cell wall degrading enzyme is added during pre-saccharification or saccharification.

49. The process of claims 44-48, wherein the yeast cell wall degrading enzyme is added during fermentation.

50. The process of claims 44-49, wherein steps (c) and (d) are carried out either simultaneously or separately/sequentially.

51. The process of claims 44-50, wherein an optionally ethanol recovery step is carried out after step (e).

52. The process of claims 44-51 wherein the milling step (a) is a dry milling step or wet milling step.

53. The process of claims 44-52, wherein the alpha-amylase used for liquefaction in step (b) is derived from a strain of the genus Bacillus or a strain of Aspergillus.

54. The process of claims 44-53, wherein the fermentation is carried out using a micro-organism capable of fermenting sugars to ethanol.

55. The process of claim 54, wherein the micro-organism is a yeast, such as derived from Saccharomyces spp., in particular Saccharomyces cerevisae.

56. The process of claims 54-55, wherein the fermentation is carried out in the presence of at least one carbohydrate-generating enzyme, in particular a glucoamylase, and further a protease.

57. The process of claim 56, wherein the protease is selected from the group of fungal proteases, such as an acid fungal protease derived from a strain of Aspergillus, in particular A. niger.

58. The process of claim 56, wherein the protease is a neutral or alkaline protease, such as a protease derived from a strain of Bacillus, in particular Alcalase®

or Neutrase®

.

34. A process of producing a fermentation product, which process comprises a fermentation step, wherein the fermentation step is carried out in the presence of one or more yeast cell wall degrading enzyme(s).

35. The process of claim 34, wherein the yeast cell wall degrading enzyme(s) is(are) selected from the group including beta-1,3-glucanase, 1,3-beta-glucanase, laminarinase, xylanase, chitinase, mannanase, alpha-1,3-glucanase (mutanase), and protease.

36. The process of claim 34-35, wherein the yeast cell wall degrading enzyme is a preparation, such as the product GLUCANEX™

derived from Trichoderma harzianum.

37. The process of claims 34-36, wherein the fermentation is carried out using a micro-organism capable of fermenting sugars or converted sugars, such as glucose or maltose.

38. The process of claim 37, wherein the micro-organism is a yeast, in particular Saccharomyces cerevisae.

39. The process of claims 34-38, wherein the material to be fermented is liquefied whole grain mash or a side stream from starch processing, in particular liquefied starch with a DE of 8-10.

40. The process of claims 34-39, wherein the fermentation is carried out in the presence of further a carbohydrate-source generating enzyme, such as glucoamylase, beta-amylase or maltogenic amylase, and/or further a protease.

41. The process of claims 34-40, wherein the protease is selected from the group of fungal proteases, such as an acid fungal protease derived from a strain of Aspergillus, in particular A. niger.

42. The process of claims 34-41, wherein the protease is a bacterial protease, such as an acid, neutral or alkaline protease, such as a protease derived from a strain of Bacillus, in particular ALCALASE®

or NEUTRASE®

.

43. The process of claims 34-42, wherein the product is ethanol or the product is beer or wine.

59. The process of any of claims 34-58 wherein further at least one carbohydrate-source generating enzyme activity and at least one alpha-amylase activity are present.

61. Use of a yeast cell wall degrading enzyme for saccharification and/or fermentation

62. Use of a yeast cell wall degrading enzyme for ethanol production.

63. A composition comprising a yeast cell wall degrading enzyme and at least one carbohydrate-generating enzyme, in particular a glucoamylase.

64. A composition of claim 63, said composition further comprising a protease, in particular an acid protease, especially an acid fungal protease.

65. The composition of claim 63 wherein further at least one alpha-amylase activity is present.

66. Use of a composition of claim 63 for saccharification and/or fermentation.

67. Use of a composition of claim 63 for ethanol production.

68. Use of a composition of claim 63 for beer or wine production.