RECOMBINANT THERMOASCUS AURANTIACUS BETA-GLUCOSIDASE VARIANTS FOR PRODUCTION OF FERMENTABLE SUGARS FROM CELLULOSIC BIOMASS

• US 20140113352A1
• Filed: 12/19/2013
• Published: 04/24/2014
• Est. Priority Date: 11/25/2009
• Status: Active Grant

• Current Assignee: Codexis, Inc.
• Original Assignee: Codexis, Inc.
• Inventors: Baidyaroy, Dipnath, Clark, Louis, Newman, Lisa M.
• Granted Patent: US 8,993,277 B2
• Time in Patent Office: Days
• US Class Current: 435/209
• CPC Class Codes: C12N 9/2445 Beta-glucosidase (3.2.1.21)
• CPC Class Codes: C12P 19/14 produced by the action of a carbohydrase (EC 3.2.x), e.g. by alpha-amylase , e.g. by cellulase, hemicellulase
• CPC Class Codes: C12Y 302/01021 Beta-glucosidase (3.2.1.21)
• CPC Class Codes: Y02E 50/10 Biofuels, e.g. bio-diesel

Abstract

The present invention provides compositions and methods for the expression of recombinant β-glucosidase variants, as well as their use in the production of fermentable sugars from cellulosic biomass.

Claims

1. A polynucleotide sequence encoding said β

-glucosidase polypeptide variant, wherein said variant β

-glucosidase comprises an amino acid sequence that is at least about 70% identical to SEQ ID NO;

2 having at least one modification or which is encoded by a nucleic acid that hybridizes under stringent conditions to the complement of SEQ ID NO;

1, wherein said variant has greater enzymatic activity than the β

-glucosidase of SEQ ID NO;

2.

3. An expression vector comprising the polynucleotide sequence of claim 1.

5. A host cell comprising the expression vector of claim 3.

7. A method for producing at least one β

-glucosidase variant comprising providing said host cell of claim 5, and culturing said host cell under conditions such that said β

-glucosidase variant is expressed.

9. The method of claim 7, further comprising the step of isolating said β

-glucosidase variant.

2. A polynucleotide sequence encoding said β

-glucosidase polypeptide variant, wherein said variant β

-glucosidase comprises an amino acid sequence that is at least about 70% identical to SEQ ID NO;

4, and having at least one modification relative to SEQ ID NO;

4 of an amino acid residue described herein, where said variant has greater enzymatic activity than SEQ ID NO;

4.

4. An expression vector comprising the polynucleotide sequence of claim 2.

6. A host cell comprising the expression vector of claim 4.

8. A method for producing at least one β

-glucosidase variant comprising providing said host cell of claim 6, and culturing said host cell under conditions such that said β

-glucosidase variant is expressed.

10. The method of claim 8, further comprising the step of isolating said β

-glucosidase variant.