Beta-glucosidase I variants with improved properties

Patent metadata

Field	Value
Publication number	US-9447400-B2
Application number	US-201013510902-A
Country	US
Kind code	B2
Filing date	Nov 19, 2010
Priority date	Nov 20, 2009
Publication date	Sep 20, 2016
Grant date	Sep 20, 2016

Abstract

The present disclosure is generally directed to enzymes and in particular beta-glucosidase variants. Also described are nucleic acids encoding beta-glucosidase variants, compositions comprising beta-glucosidase variants, methods of using beta-glucosidase variants, and methods of identifying additional useful beta-glucosidase variants.

First claim

What is claimed is:

1. A beta-glucosidase 1 (BGL1) variant having at least 80% amino acid sequence identity to SEQ ID NO: 3 wherein the variant comprises a substitution corresponding to position Q226 in SEQ ID NO: 3 with W or Y, a substitution corresponding to N263 in SEQ ID NO: 3 with C, S, or T, a substitution corresponding to K345 in SEQ ID NO: 3 with E, a substitution corresponding to P661 in SEQ ID NO: 3 with E, F, L, I, or Q, or a substitution corresponding to G372 of SEQ ID NO: 3 with A.

2. The BGL1 variant of claim 1 wherein the variant comprises a substitution corresponding to G372 of SEQ ID NO: 3 with A.

3. A beta-glucosidase 1 (BGL1) variant having at least 80% amino acid sequence identity to SEQ ID NO: 3 wherein the BGL1 variant comprises a substitution corresponding to position Q226 in SEQ ID NO: 3 with W or Y, a substitution corresponding to N263 in SEQ ID NO: 3 of C, S, or T, a substitution corresponding to P661 in SEQ ID NO: 3 of F, L, or Q, a substitution corresponding to K345 in SEQ ID NO: 3 of E, a substitution corresponding to S683 in SEQ ID NO: 3 with W, or a substitution corresponding to G372 in SEQ ID NO: 3 with A.

4. A beta-glucosidase 1 (BGL1) variant having at least 80% amino acid sequence identity to SEQ ID NO: 3 wherein the BGL1 variant comprises a substitution corresponding to position Q226 in SEQ ID NO: 3 with W or Y, a substitution corresponding to N263 in SEQ ID NO: 3 of C or S, a substitution corresponding to P661 in SEQ ID NO: 3 of F, L, or Q, a substitution corresponding to S683 in SEQ ID NO: 3 with W, a substitution corresponding to K345 in SEQ ID NO: 3 with E or a substitution corresponding to G372 in SEQ ID NO: 3 with A.

5. A BGL1 variant according to claim 1 wherein the BGL1 variant comprises a substitution corresponding to N263 in SEQ ID NO: 3 with T.

6. A beta-glucosidase 1 (BGL1) variant having at least 80% amino acid sequence identity to SEQ ID NO: 3 wherein the BGL1 variant comprises a substitution corresponding to position Q226 in SEQ ID NO: 3 with W or Y, a substitution corresponding to N263 in SEQ ID NO: 3 of C or S, a substitution corresponding to P661 in SEQ ID NO: 3 of F, L, or Q, a substitution corresponding to K345 in SEQ ID NO: 3 of E, or a substitution corresponding to S683 in SEQ ID NO: 3 with W.

7. A composition comprising the BGL1 variant of claim 1 .

8. The composition of claim 7 wherein the composition is enriched in the BGL1 variant.

9. An isolated nucleic acid encoding any one of the BGL1 variants of claim 1 .

10. An expression vector comprising the nucleic acid of claim 9 .

11. The expression vector of claim 10 wherein the isolated nucleic acid is operably linked to a regulatory sequence.

12. A host cell comprising the nucleic acid of claim 9 .

13. A method for producing a BGL1 variant, comprising culturing the host cell of claim 12 in a culture medium under suitable conditions to produce the variant.

14. A method of converting biomass to sugars comprising contacting the biomass with a BGL1 variant of claim 1 .

15. A composition comprising the BGL1 variant of claim 3 .

16. The composition of claim 15 wherein the composition is enriched in the BGL1 variant.

17. A composition comprising the BGL1 variant of claim 4 .

18. The composition of claim 17 wherein the composition is enriched in the BGL1 variant.

19. A composition comprising the BGL1 variant of claim 6 .

20. The composition of claim 19 wherein the composition is enriched in the BGL1 variant.

Assignees

• Bott Richard R
• Kaper Thijs
• Kelemen Bradley
• Goedegebuur Frits
• Hommes Ronaldus Wilhelmus
• Kralj Slavko
• Kruithof Paulien
• Nikolaev Igor
• Van Der Kley Wilhelmus Antonious Hendricus
• Van Lieshout Johannes Franciscus Thomas
• Van Stigt Thans Sander
• Vogtentanz Gudrun
• Sandgren Mats
• Danisco Us Inc

Inventors

• Bott Richard R
• Kaper Thijs
• Kelemen Bradley
• Goedegebuur Frits
• Hommes Ronaldus Wilhelmus
• Kralj Slavko
• Kruithof Paulien
• Nikolaev Igor
• Van Der Kley Wilhelmus Antonious Hendricus
• Van Lieshout Johannes Franciscus Thomas
• Van Stigt Thans Sander
• Vogtentanz Gudrun
• Sandgren Mats

Classifications

• C12N9/2445 Primary

  Beta-glucosidase (3.2.1.21) · CPC title

• C12Y302/01021

  Beta-glucosidase (3.2.1.21) · CPC title

• C12P19/02

  Monosaccharides (2-ketogulonic acid C12P7/60) · 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

• C12N15/52

  Genes encoding for enzymes or proenzymes · CPC title