Ionic liquid-tolerant cellulase enzymes

Patent metadata

Field	Value
Publication number	US-9803182-B2
Application number	US-201514612201-A
Country	US
Kind code	B2
Filing date	Feb 2, 2015
Priority date	Feb 3, 2014
Publication date	Oct 31, 2017
Grant date	Oct 31, 2017

Abstract

The present invention provides ionic liquid-tolerant cellulases and method of producing and using such cellulases. The cellulases of the invention are useful in saccharification reactions using ionic liquid treated biomass.

First claim

What is claimed is:

1. A composition comprising: (i) an ionic-liquid-tolerant β-glucosidase comprising an amino acid sequence having at least 99% sequence identity to the amino acid sequence of SEQ ID NO:2 and having β-glucosidase activity; (ii) an ionic liquid that has an imidazolium cation: and (iii) a lignocellulosic biomass.

2. The composition of claim 1 , wherein the ionic liquid is [C 2 mim][OAc].

3. The composition of claim 1 , wherein the ionic-liquid-tolerant β-glucosidase comprises the amino acid sequence of SEQ ID NO:2.

4. The composition of claim 1 , further comprising soluble sugars.

5. The composition of claim 3 , wherein the ionic liquid is [C 2 mim][OAc].

6. The composition of claim 1 , further comprising at least one ionic-liquid-tolerant endoglucanase comprising an amino acid sequence selected from the group consisting of SEQ ID NOS:16-26.

7. A method of increasing the yield of soluble sugar from a lignocellulosic biomass, the method comprising incubating lignocellulosic biomass with a composition comprising an ionic-liquid-tolerant β-glucosidase and an ionic liquid having an imidazolium cation in an enzymatic hydrolysis reaction, wherein the ionic-liquid-tolerant β-glucosidase comprises an amino acid sequence having at least 99% sequence identity to the amino acid sequence of SEQ ID NO:2 and has β-glucosidase activity.

8. The method of claim 7 , wherein the biomass is pretreated with ionic liquid.

9. The method of claim 7 , wherein the ionic liquid is [C 2 mim][OAc].

10. The composition of claim 6 , wherein the ionic liquid-tolerant endoglucanase comprises an amino acid sequence selected from the group consisting of SEQ ID NO:16, 17, 18, 19, 20, and 21.

11. The composition of claim 6 , wherein the ionic liquid-tolerant endoglucanase comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 17, 18, and 21.

12. The method of claim 7 , wherein the ionic-liquid-tolerant β-glucosidase comprises the amino acid sequence of SEQ ID NO:2.

13. The method of claim 7 , wherein the enzymatic hydrolysis reaction further comprises at least one ionic-liquid-tolerant endoglucanase comprising an amino acid sequence selected from the group consisting of SEQ ID NOS: 16-26.

14. The method of claim 13 , wherein the ionic-liquid-tolerant endoglucanase comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 16, 17, 18, 19, 20, and 21.

15. The method of claim 13 , wherein the ionic-liquid-tolerant endoglucanase comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 17, 18, and 21.

Assignees

• Univ California
• Sandia Corp

Inventors

• Gladden John
• Park Joshua
• Singer Steven
• Simmons Blake
• Sale Ken

Classifications

• C12N9/2437

  Cellulases (3.2.1.4; 3.2.1.74; 3.2.1.91; 3.2.1.150) · CPC title

• C12P19/02

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

• C12N9/2445 Primary

  Beta-glucosidase (3.2.1.21) · 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

• C12Y302/01004

  Cellulase (3.2.1.4), i.e. endo-1,4-beta-glucanase · CPC title