Polynucleotides encoding beta-glucosidase variants

• US 8,993,277 B2
• Filed: 12/19/2013
• Issued: 03/31/2015
• 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.
• Primary Examiner(s): ROBINSON, HOPE A
• Application Number:
  US14/134,839
• Publication Number:
  US 20140113352A1
• Time in Patent Office: 467 Days
• Field of Search: None
• US Class Current: 435/105
• 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 recombinant polynucleotide sequence encoding a recombinant β

-glucosidase polypeptide variant, wherein said variant β

-glucosidase comprises an amino acid sequence that is at least 95% identical to SEQ ID NO;

2, wherein said variant comprises a substitution set selected from the group consisting of;

D204G+K292I+E345V+Y747C;

H685Y+P791T;

T151S+Y642N+N651K;

M1T+K55R+K101R+T151S+R331K+Y332C+K343R+N356S+S409N+Y642N;

M1T+K101R+T151S+K292E +K343R+S409N+Y642N+P740S;

M 1T+T151S+K343R+S409N+A479V+Y642N+Y680F;

L150V+T151S+K343R+S409N+K457R+Y642N+N651 K; and

S 87N+T151S +F288Y+Y642N+N651 K, and wherein said variant has greater enzymatic activity than the 13-glucosidase of SEQ ID NO;

2.

2. An expression vector comprising the recombinant polynucleotide sequence of claim 1.

3. A host cell comprising the expression vector of claim 2.

4. A method for producing at least one β

-glucosidase variant comprising providing said host cell of claim 3, and culturing said host cell under conditions such that said β

-glucosidase variant is expressed.

5. The method of claim 4, further comprising the step of isolating said β

-glucosidase variant.