Enhanced processive cellulases

Patent metadata

Field	Value
Publication number	US-10036051-B2
Application number	US-201715594031-A
Country	US
Kind code	B2
Filing date	May 12, 2017
Priority date	Dec 16, 2011
Publication date	Jul 31, 2018
Grant date	Jul 31, 2018

Abstract

Nucleic acid sequences encoding chimeric polypeptides that exhibit enhanced cellulase activities are disclosed herein. These nucleic acids may be expressed in hosts such as fungi, which in turn may be cultured to produce chimeric polypeptides. Also disclosed are chimeric polypeptides and their use in the degradation of cellulosic materials.

First claim

We claim:

1. An isolated chimeric fungal Cel7A polypeptide that has a cellulase activity at least 1.5-fold greater than the wild-type Cel7A polypeptide, comprising a catalytic domain (CD), a carbohydrate-binding molecule (CBM) and a linker domain, wherein at least one of the domains is from a Penicillium funiculosum Cel7A polypeptide and at least one of the domains is from a Trichoderma reesei Cel7A polypeptide; and wherein each domain is from the Penicillium funiculosum or Trichoderma reesei Cel7A polypeptides.

2. The isolated chimeric fungal Cel7A polypeptide of claim 1 , wherein the chimeric fungal Cel7A polypeptide has a cellulase activity of at least 2-fold greater than the wild-type Cel7A polypeptide.

3. The isolated chimeric fungal Cel7A polypeptide of claim 1 , wherein the chimeric fungal Cel7A polypeptide comprises the catalytic domain (CD) from the Penicillium funiculosum Cel7A polypeptide.

4. The isolated chimeric fungal Cel7A polypeptide of claim 3 , wherein the chimeric fungal Cel7A polypeptide further comprises the carbohydrate-binding molecule (CBM) from the Trichoderma reesei Cel7A peptide.

5. The isolated chimeric fungal Cel7A polypeptide of claim 4 , wherein the chimeric fungal Cel7A polypeptide further comprises the linker domain from the Penicillium funiculosum Cel7A polypeptide.

6. The isolated chimeric fungal Cel7A polypeptide of claim 4 , wherein the chimeric fungal Cel7A polypeptide comprises the linker domain from the Trichoderma reesei Cel7A polypeptide.

7. The isolated chimeric fungal Cel7A polypeptide of claim 3 , wherein the chimeric fungal Cel7A polypeptide further comprises the linker domain from the Trichoderma reesei Cel7A polypeptide.

8. The isolated chimeric fungal Cel7A polypeptide of claim 7 , wherein the chimeric fungal Cel7A polypeptide further comprises the carbohydrate-binding molecule (CBM) from the Penicillium funiculosum Cel7A polypeptide.

9. The isolated chimeric fungal Cel7A polypeptide of claim 1 , wherein the chimeric fungal Cel7A polypeptide comprises the catalytic domain (CD) from the Trichoderma reesei Cel7A polypeptide.

10. The isolated chimeric fungal Cel7A polypeptide of claim 9 , wherein the chimeric fungal Cel7A polypeptide further comprises the linker domain from the Penicillium funiculosum Cel7A polypeptide.

11. The isolated chimeric fungal Cel7A polypeptide of claim 10 , wherein the chimeric fungal Cel7A polypeptide further comprises the carbohydrate-binding molecule (CBM) form the Penicillium funiculosum Cel7A polypeptide.

12. The isolated chimeric fungal Cel7A polypeptide of claim 10 , wherein the chimeric fungal Cel7A polypeptide further comprises the carbohydrate-binding molecule (CBM) from the Trichoderma reesei Cel7A polypeptide.

13. The isolated chimeric fungal Cel7A polypeptide of claim 9 , wherein the chimeric fungal Cel7a polypeptide further comprises the carbohydrate-binding molecule (CBM) from the Penicillium funiculosum Cel7A polypeptide.

14. The isolated chimeric fungal Cel7A polypeptide of claim 13 , wherein the chimeric fungal Cel7A polypeptide further comprises the linker domain from the Trichoderma reesei Cel7A polypeptide.

15. A method for degrading cellulose or lignocellulose biomass, comprising contacting the cellulose or lignocellulosic biomass with the isolated chimeric Cel7A polypeptide according to claim 1 .

16. A method for producing a biofuel from lignocellulosic biomass, comprising:
a) contacting the lignocellulosic biomass with an enzyme cocktail comprising the isolated chimeric fungal Cel7A polypeptide according to claim 1 to generate sugars; and
b) converting the sugars to a biofuel by fermentation.

17. The method of claim 16 , wherein the enzyme cocktail further comprises an endoglucanase, a β-glucosidase, or both.

Assignees

• Alliance Sustainable Energy

Inventors

• Adney William S
• Beckham Gregg T
• Jarvis Eric
• Himmel Michael E
• Decker Stephen R
• Linger Jeffrey G
• Podkaminer Kara
• Baker John O
• Taylor Ii Larry
• Xu Qi
• Singh Arjun

Classifications

• C12Y302/01004

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

• C10L2290/26

  Composting, fermenting or anaerobic digestion fuel components or materials from which fuels are prepared · CPC title

• C12P19/14 Primary

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

• C12N9/2437

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

• C12Y302/01021

  Beta-glucosidase (3.2.1.21) · CPC title