Recombinant Thermoascus aurantiacus β-glucosidase variants

• US 8,652,813 B2
• Filed: 11/19/2010
• Issued: 02/18/2014
• Est. Priority Date: 11/25/2009
• Status: Expired due to Fees

• Current Assignee: Codexis, Inc.
• Original Assignee: Codexis, Inc.
• Inventors: Baidyaroy, Dipnath, Clark, Louis, Newman, Lisa M.
• Primary Examiner(s): ROBINSON, HOPE A
• Application Number:
  US12/950,150
• Publication Number:
  US 20110124058A1
• Time in Patent Office: 1,187 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 variant β

-glucosidase, comprising a substitution set selected from the group consisting of;

T151S-Y642N-N651K, K101R-T151S-K343R-N356S-S409N-Y642N-N651K, M1T-K55R-K101R-T151S-R331K-Y332C-K343R-N356S-S409N-Y642N, M1T-K101R-T151S-K292E-K343R-S409N-Y642N-P740S, M1T-T151S-K343R-S409N-A479V-Y642N-Y680F, L150V-T151S-K343R-S409N-K457R-Y642N-N651K, and S87N-T151S-F288Y-Y642N-N651K of SEQ ID NO;

4, wherein said recombinant variant β

-glucosidase has greater enzymatic activity than the amino acid sequence of SEQ ID NO;

4.

2. The recombinant variant β

-glucosidase of claim 1, wherein the polynucleotide sequence encoding said variant comprises a polynucleotide sequence having at least one base change selected from t1044c, t1656a, t2052c, and/or a2520g wherein said bases are found in the polynucleotide sequence of SEQ ID NO;

3.

3. A composition comprising the recombinant variant β

-glucosidase of claim 1.

4. The composition of claim 3, further comprising at least one additional enzyme.

5. The composition of claim 4, wherein said at least one additional enzyme is a cellulase.

6. The composition of claim 4, wherein said at least one additional enzyme is an additional recombinant variant β

-glucosidase of SEQ ID NO;

4.