Processes for producing ethanol

Patent metadata

Field	Value
Publication number	US-11566266-B2
Application number	US-202017036601-A
Country	US
Kind code	B2
Filing date	Sep 29, 2020
Priority date	Dec 22, 2010
Publication date	Jan 31, 2023
Grant date	Jan 31, 2023

Abstract

The present invention relates to processes for producing fermentation products from starch-containing material, wherein a thermostable alpha-amylase and optionally a thermostable protease are present and/or added during liquefaction. The invention also relates to a composition suitable for use in a process of the invention.

First claim

The invention claimed is:

1. A process for producing fuel ethanol from a starch-containing material comprising:
i) liquefying a starch-containing material at a temperature above the initial gelatinization temperature using:
an alpha-amylase; and
a protease having at least 80% sequence identity to the amino acid sequence set forth in SEQ ID NO: 13 and a thermostability value of more than 20% determined as Relative Activity at 80° C./70° C., wherein the starch-containing material comprises corn;
ii) saccharifying the liquefied material obtained in step i) using a glucoamylase; and
iii) fermenting the saccharified material obtained in step (ii) using a yeast fermenting organism to produce fuel ethanol.

2. The process of claim 1 , wherein the alpha-amylase is a variant of a Bacillus alpha-amylase.

3. The process of claim 1 , wherein the alpha-amylase is a variant of a Bacillus stearothermophilus alpha-amylase.

4. The process of claim 1 , wherein the alpha-amylase has at least 80%, but less than 100% identity to the mature part of the polypeptide of SEQ ID NO: 1.

5. The process of claim 4 , wherein the alpha-amylase is truncated.

6. The process of claim 1 , wherein the alpha-amylase is a truncated Bacillus stearothermophilus alpha-amylase with the mutations selected from the group consisting of:
I181*+G182*; and
I181*+G182*+N193F; wherein each position corresponds to a position in the amino acid sequence set forth in SEQ ID NO: 1.

7. The process of claim 1 , wherein the alpha-amylase has a T½ (min) at pH 4.5, 85° C., 0.12 mM CaCl2) of at least 10.

8. The process of claim 1 , wherein the alpha-amylase has a T½ (min) at pH 4.5, 85° C., 0.12 mM CaCl2) of at least 15.

9. The process of claim 1 , wherein the alpha-amylase has a T½ (min) at pH 4.5, 85° C., 0.12 mM CaCl2) of at least 20.

10. The process of claim 1 , wherein the alpha-amylase has a T½ (min) at pH 4.5, 85° C., 0.12 mM CaCl2) of at least 25.

11. The process of claim 1 , wherein the alpha-amylase has a T½ (min) at pH 4.5, 85° C., 0.12 mM CaCl2) of at least 30.

12. The process of claim 1 , wherein the alpha-amylase has a T½ (min) at pH 4.5, 85° C., 0.12 mM CaCl2) of at least 40.

13. The process of claim 1 , wherein the alpha-amylase has a T½ (min) at pH 4.5, 85° C., 0.12 mM CaCl2) of at least 50.

14. The process of claim 1 , wherein the alpha-amylase has a T½ (min) at pH 4.5, 85° C., 0.12 mM CaCl2) of at least 60.

15. The process of claim 1 , wherein the alpha-amylase has a T½ (min) at pH 4.5, 85° C., 0.12 mM CaCl2) of at least 70.

16. The process of claim 1 , wherein the protease is a serine protease.

17. The process of claim 1 , wherein the protease is from a strain of Pyrococcus.

18. The process of claim 1 , wherein the protease has at least 85% sequence identity to the amino acid sequence set forth in SEQ ID NO: 13.

19. The process of claim 1 , wherein the protease has as at least 90% sequence identity to SEQ ID NO: 13.

20. The process of claim 1 , wherein the protease has as at least 95% sequence identity to SEQ ID NO: 13.

21. The process of claim 1 , wherein the protease has a thermostability value of more than 40% determined as Relative Activity at 80° C./70° C.

22. The process of claim 1 , wherein the protease has a thermostability value of more than 50% determined as Relative Activity at 80° C./70° C.

23. The process of claim 1 , wherein the protease has a thermostability value of more than 60% determined as Relative Activity at 80° C./70° C.

24. The process of claim 1 , wherein the protease has a thermostability value of more than 70% determined as Relative Activity at 80° C./70° C.

25. The process of claim 1 , wherein the protease has a thermostability value of more than 80% determined as Relative Activity at 80° C./70° C.

26. The process of claim 1 , wherein the protease has a thermostability value of more than 90% determined as Relative Activity at 80° C./70° C.

27. The process of claim 1 , wherein the protease has a thermostability value of more than 100% determined as Relative Activity at 80° C./70° C.

28. The process of claim 1 , wherein the protease has a thermostability value of more than 110% determined as Relative Activity at 80° C./70° C.

29. The process of claim 1 , wherein the protease has a thermostability value of more than 12% determined as Relative Activity at 85° C./70° C.

30. The process of claim 1 , wherein the protease has a thermostability value of more than 14% determined as Relative Activity at 85° C./70° C.

31. The process of claim 1 , wherein the protease has a thermostability value of more than 16% determined as Relative Activity at 85° C./70° C.

32. The process of claim 1 , wherein the protease has a thermostability value of more than 18% determined as Relative Activity at 85° C./70° C.

33. The process of claim 1 , wherein the protease has a thermostability value of more than 20% determined as Relative Activity at 85° C./70° C.

34. The process of claim 1 , wherein the protease has a thermostability value of more than 25% determined as Relative Activity at 85° C./70° C.

35. The process of claim 1 , wherein the protease has a thermostability value of more than 20% determined as Remaining Activity at 80° C.

36. The process of claim 1 , wherein the protease has a thermostability value of more than 30% determined as Remaining Activity at 80° C.

37. The process of claim 1 , wherein the protease has a thermostability value of more than 40% determined as Remaining Activity at 80° C.

38. The process of claim 1 , wherein the protease has a thermostability value of more than 50% determined as Remaining Activity at 80° C.

39. The process of claim 1 , wherein the protease has a thermostability value of more than 60% determined as Remaining Activity at 80° C.

40. The process of claim 1 , wherein the protease has a thermostability value of more than 70% determined as Remaining Activity at 80° C.

41. The process of claim 1 , wherein the protease has a thermostability value of more than 80% determined as Remaining Activity at 80° C.

42. The process of claim 1 , wherein the protease has a thermostability value of more than 90% determined as Remaining Activity at 80° C.

43. The process of claim 1 , wherein the protease has a thermostability value of more than 20% determined as Remaining Activity at 84° C.

44. The process of claim 1 , wherein the protease has a thermostability value of more than 30% determined as Remaining Activity at 84° C.

45. The process of claim 1 , wherein the protease has a thermostability value of more than 40% determined as Remaining Activity at 84° C.

46. The process of claim 1 , wherein the protease has a thermostability value of more than 50% determined as Remaining Activity at 84° C.

47. The process of claim 1 , wherein the protease has a thermostability value of more than 60% determined as Remaining Activity at 84° C.

48. The process of claim 1 , wherein the protease has a thermostability value of more than 70% determined as Remaining Activity

Assignees

• Novozymes As

Inventors

• Deinhammer Randy
• Clark Suzanne
• Quiros Mauricio
• Matthews John
• Hjulmand Anne Glud
• Soong Chee Leong
• Matsui Tomoko
• Takagi Shinobu

Classifications

• C12P19/20

  produced by the action of an exo-1,4 alpha-glucosidase, e.g. dextrose · CPC title

• C12P19/14

  produced by the action of a carbohydrase {(EC 3.2.x)}, e.g. by alpha-amylase {, e.g. by cellulase, hemicellulase} · CPC title

• C12P7/06 Primary

  Ethanol, i.e. non-beverage · CPC title

• C12P19/02

  Monosaccharides (2-ketogulonic acid C12P7/60) · CPC title

• C12Y304/24

  Metalloendopeptidases (3.4.24) · CPC title