Compositions comprising a low pH alpha-amylase and a thermostable protease for liquefaction

Patent metadata

Field	Value
Publication number	US-12227785-B2
Application number	US-202318492525-A
Country	US
Kind code	B2
Filing date	Oct 23, 2023
Priority date	Dec 22, 2010
Publication date	Feb 18, 2025
Grant date	Feb 18, 2025

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 composition comprising an alpha-amylase and a protease, wherein i) the alpha-amylase has at least 85% identity, but less than 100% identity, to the polypeptide of SEQ ID NO: 1 and has a T½ (min) at pH 4.5, 85 degrees centigrade, 0.12 mM CaCl 2 of at least 10; and
ii) the protease has at least 85% identity, but less than 100% identity, to the polypeptide of SEQ ID NO: 13, wherein the protease has a thermostability value of more than 50 percent determined as Relative Activity at 80 degrees centigrade/70 degrees centigrade.

2. The composition of claim 1 , wherein the alpha-amylase has a double deletion in I181+G182.

3. The composition of claim 1 , wherein the alpha-amylase has a substitution N193F.

4. The composition of claim 1 , wherein the alpha-amylase has a double deletion in I181+G182 and a substitution N193F.

5. The composition of claim 1 , wherein the alpha-amylase has a double deletion in I181+G182 and a T½ (min) at pH 4.5, 85 degrees centigrade, 0.12 mM CaCl 2 of at least 20, and wherein the protease has a thermostability value of more than 70 percent determined as Relative Activity at 80 degrees centigrade/70 degrees centigrade.

6. The composition of claim 1 , wherein the alpha-amylase has a double deletion in I181+G182, a substitution N193F, and a T½ (min) at pH 4.5, 85 degrees centigrade, 0.12 mM CaCl 2 of at least 20, and wherein the protease has a thermostability value of more than 70 percent determined as Relative Activity at 80 degrees centigrade/70 degrees centigrade.

7. The composition of claim 1 , wherein the alpha-amylase has a double deletion in I181+G182, a substitution N193F, and a T½ (min) at pH 4.5, 85 degrees centigrade, 0.12 mM CaCl 2 of at least 25, and wherein the protease has a thermostability value of more than 90 percent determined as Relative Activity at 80 degrees centigrade/70 degrees centigrade.

8. A composition comprising an alpha-amylase and a protease, wherein
i) the alpha-amylase has at least 90% identity, but less than 100% identity, to the polypeptide of SEQ ID NO: 1 and has a T½ (min) at pH 4.5, 85 degrees centigrade, 0.12 mM CaCl 2 of at least 10; and
ii) the protease has at least 90% identity, but less than 100% identity, to the polypeptide of SEQ ID NO: 13, wherein the protease has a thermostability value of more than 50 percent determined as Relative Activity at 80 degrees centigrade/70 degrees centigrade.

9. The composition of claim 8 , wherein the alpha-amylase has a double deletion in I181+G182.

10. The composition of claim 8 , wherein the alpha-amylase has a substitution N193F.

11. The composition of claim 8 , wherein the alpha-amylase has a double deletion in I181+G182 and a substitution N193F.

12. The composition of claim 8 , wherein the alpha-amylase has a double deletion in I181+G182 and a T½ (min) al pH 4.5, 85 degrees centigrade, 0.12 mM CaCl 2 of at least 20, and wherein the protease has a thermostability value of more than 70 percent determined as Relative Activity at 80 degrees centigrade/70 degrees centigrade.

13. The composition of claim 8 , wherein the alpha-amylase has a double deletion in I181+G182, a substitution N193F, and a T½ (min) at pH 4.5, 85 degrees centigrade, 0.12 mM CaCl 2 of at least 20, and wherein the protease has a thermostability value of more than 70 percent determined as Relative Activity at 80 degrees centigrade/70 degrees centigrade.

14. The composition of claim 8 , wherein the alpha-amylase has a double deletion in I181+G182, a substitution N193F, and a T½ (min) at pH 4.5, 85 degrees centigrade, 0.12 mM CaCl 2 of at least 25, and wherein the protease has a thermostability value of more than 90 percent determined as Relative Activity at 80 degrees centigrade/70 degrees centigrade.

15. A composition comprising an alpha-amylase and a protease, wherein
i) the alpha-amylase has at least 95% identity, but less than 100% identity, to the polypeptide of SEQ ID NO: 1 and has a T½ (min) at pH 4.5, 85 degrees centigrade, 0.12 mM CaCl 2 of at least 10; and
ii) the protease has at least 95% identity, but less than 100% identity, to the polypeptide of SEQ ID NO: 13, wherein the protease has a thermostability value of more than 50 percent determined as Relative Activity at 80 degrees centigrade/70 degrees centigrade.

16. The composition of claim 15 , wherein the alpha-amylase has a double deletion in I181+G182.

17. The composition of claim 15 , wherein the alpha-amylase has a substitution N193F.

18. The composition of claim 15 , wherein the alpha-amylase has a double deletion in I181+G182 and a substitution N193F.

19. The composition of claim 15 , wherein the alpha-amylase has a double deletion in I181+G182 and a T½ (min) at pH 4.5, 85 degrees centigrade, 0.12 mM CaCl 2 of at least 20, and wherein the protease has a thermostability value of more than 70 percent determined as Relative Activity at 80 degrees centigrade/70 degrees centigrade.

20. The composition of claim 15 , wherein the alpha-amylase has a double deletion in I181+G182, a substitution N193F, and a T½ (min) at pH 4.5, 85 degrees centigrade, 0.12 mM CaCl 2 of at least 20, and wherein the protease has a thermostability value of more than 70 percent determined as Relative Activity at 80 degrees centigrade/70 degrees centigrade.

21. The composition of claim 15 , wherein the alpha-amylase has a double deletion in I181+G182, a substitution N193F, and a T½ (min) at pH 4.5, 85 degrees centigrade, 0.12 mM CaCl 2 of at least 25, and wherein the protease has a thermostability value of more than 90 percent determined as Relative Activity at 80 degrees centigrade/70 degrees centigrade.

Assignees

• Novozymes As

Inventors

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

Classifications

• Y02E50/10

  Biofuels, e.g. bio-diesel · CPC title

• C12Y302/01003

  Glucan 1,4-alpha-glucosidase (3.2.1.3), i.e. glucoamylase · CPC title

• 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

• C12P19/02

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