Processes for producing ethanol

Patent metadata

Field	Value
Publication number	US-10947567-B2
Application number	US-201715727952-A
Country	US
Kind code	B2
Filing date	Oct 9, 2017
Priority date	Dec 22, 2010
Publication date	Mar 16, 2021
Grant date	Mar 16, 2021

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 ethanol from starch-containing material comprising:
i) liquefying the starch-containing material at a temperature above 80-90° C. using:
a thermostable bacterial alpha-amylase and
a thermostable bacterial protease having a thermostability value of more than 20% determined as Relative Activity at 80° C./70° C.;

ii) saccharifying the liquefied material obtained in step i) using a glucoamylase; and
iii) fermenting the saccharified material obtained in step (ii) using a fermenting organism.

2. 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.

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 variant has at least 80%, but less than 100% identity to the mature part of the polypeptide of SEQ ID NO: 1 herein.

5. The process of claim 4 , wherein the alpha-amylase is truncated to have 491 amino acids.

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*+N193F+V59A+Q89R+E129V+K177L+R179E+H208Y+K220P+N224L+Q254S;
I181*+G182*+N193F+E129V+K177L+R179E; and
I181*+G182*+N193F+E129V+K177L+R179E+K220P+N224L+S242Q+Q254S; 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 protease is a serine protease.

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

9. The process of claim 1 , wherein the protease is the Pyrococcus furiosus protease of SEQ ID NO: 13, or one having at least 90% sequence identity thereto.

10. The process of claim 9 , wherein the protease has as at least 95% sequence identity to SEQ ID NO: 13.

11. The process of claim 1 , further wherein a thermostable glucoamylase is present and/or added during liquefaction step i).

12. The process of claim 11 , wherein the glucoamylase has a heat stability at 85° C., pH 5.3, of at least 30%.

13. The process of claim 11 , wherein the glucoamylase is from a strain of the genus Penicillium.

14. The process of claim 13 , wherein the glucoamylase is a Penicillium oxalicum glucoamylase having the amino acid sequence of SEQ ID NO: 9 or 14.

15. The process of claim 14 , wherein the glucoamylase has a K79V substitution in SEQ ID NO: 9 or 14 (using the mature sequence shown in SEQ ID NO: 14 for numbering).

16. The process of claim 14 , wherein the glucoamylase has at least 80% identity to the mature polypeptide shown in SEQ ID NO: 9 or 14 herein.

17. The process of claim 1 , wherein a pullulanase is present during liquefaction and/or saccharification.

18. The process of claim 1 , wherein the glucoamylase is of fungal origin.

19. The process of claim 18 , wherein the glucoamylase is from a strain selected from the group consisting of Aspergillus, Trichoderma, Talaromyces, Pycnoporus , and Gloeophyllum.

20. The process of claim 1 , wherein the ethanol comprises fuel ethanol.

21. The process of claim 1 , wherein the starch-containing material comprises corn.

22. The process of claim 1 , wherein steps (ii) and (iii) are performed sequentially.

23. The process of claim 1 , wherein steps (ii) and (iii) are performed simultaneously.

24. The process of claim 1 , wherein the temperature in step i) is from 80° C. to 95° C.

25. The process of claim 1 , wherein the fermenting organism is yeast.

26. The process of claim 25 , wherein the yeast is a strain of Saccharomyces cerevisiae.

27. The process of claim 1 , wherein:
(i) the starch-containing material is corn;
(ii) the fermentation product is fuel ethanol;
(iii) the alpha-amylase has a T½ (min) at pH 4.5, 85° C., 0.12 mM CaCl2) of at least 10; and
(iv) the protease has a thermostability value of more than 90% determined as Relative Activity at 80° C./70° C.

28. The process of claim 1 , wherein:
(i) the starch-containing material is corn;
(ii) the fermentation product is fuel ethanol;
(iii) the alpha-amylase has a T½ (min) at pH 4.5, 85° C., 0.12 mM CaCl2) of at least 10;
(iv) the protease has a thermostability value of more than 90% determined as Relative Activity at 80° C./70° C.;
(v) the temperature during liquefying step i) is between 80° C.-95° C.;
(vi) the pH during liquefying step i) is from 4.5-5.0;
(vii) the glucomylase is of fungal origin;
(viii) the fermenting organism is a yeast; and
(ix) the saccharifying step (ii) and fermenting step (iii) are carried out as a simultaneous saccharification and fermentation.

29. The process of claim 1 , wherein:
(i) the starch-containing material is corn;
(ii) the fermentation product is fuel ethanol;
(iii) the alpha-amylase has a T½ (min) at pH 4.5, 85° C., 0.12 mM CaCl2) of at least 20;
(iv) the protease has a thermostability value of more than 90% determined as Relative Activity at 80° C./70° C.;
(v) the temperature during liquefying step i) is between 80° C.-95° C.;
(vi) the pH during liquefying step i) is from 4.5-5.0;
(vii) the glucomylase is of fungal origin;
(viii) the fermenting organism is a Saccharomyces yeast; and
(ix) the saccharifying step (ii) and fermenting step (iii) are carried out as a simultaneous saccharification and fermentation.

30. The process of claim 1 , wherein:
(i) the starch-containing material is corn;
(ii) the fermentation product is fuel ethanol;
(iii) the alpha-amylase has a T½ (min) at pH 4.5, 85° C., 0.12 mM CaCl2) of at least 20;
(iv) the protease has a thermostability value of more than 90% determined as Relative Activity at 80° C./70° C.;
(v) the temperature during liquefying step i) is between 80° C.-90° C.;
(vi) the pH during liquefying step i) is from 4.5-5.0;
(vii) the glucomylase is of fungal origin;
(viii) the fermenting organism is a Saccharomyces yeast; and
(ix) the saccharifying step (ii) and fermenting step (iii) are carried out as a simultaneous saccharification and fermentation.

31. The process of claim 1 , wherein:
(i) the starch-containing material is corn;
(ii) the fermentation product is fuel ethanol;
(iii) the alpha-amylase is a bacterial alpha-amylase derived from the genus Bacillus having a T½ (min) at pH 4.5, 85° C., 0.12 mM CaCl2) of at least 20;
(iv) the protease has a thermostability value of more than 95% determined as Relative Activity at 80° C./70° C.;
(v) the temperature during liquefying step i) is between 80° C.-95° C.;
(vi) the pH during liquefying step i) is from 4.5-5.0;
(vii) the glucomylase is of fungal origin;
(viii) the fermenting organism is a Saccharomyces yeast; and
(ix) the saccharifying step (ii) and fermenting step (iii) are carried out as a simultaneous saccharification and fermentation.

32. The process of claim 1 , wherein:
(i) the starch-containing material is corn;
(ii) the fermentation product is fuel ethanol;
(iii) the alpha-amylase is a bacterial alpha-amylase derived from the genus Bacillus having a T½ (min) at pH 4.5, 85° C., 0.12 mM CaCl2) of at least 20;
(iv) the protease has a thermostability value of more than 95% determined as Relative Activity at 80° C./70° C.;
(v) the temp

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

• C12Y304/24

  Metalloendopeptidases (3.4.24) · CPC title

• C12Y302/01001

  Alpha-amylase (3.2.1.1) · CPC title

• C12P7/06 Primary

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

• C12P19/02

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