Polypeptides having glucoamylase activity and method of producing the same

Patent metadata

Field	Value
Publication number	US-9677058-B2
Application number	US-201214366214-A
Country	US
Kind code	B2
Filing date	Dec 20, 2012
Priority date	Dec 22, 2011
Publication date	Jun 13, 2017
Grant date	Jun 13, 2017

Abstract

The present invention relates to polypeptides having glucoamylase activity with improved properties and to compositions comprising these polypeptides suitable for use in the production of a food, beverage (e.g. beer), feed, or biofuel. Also described is an improved and cost-effective process for isolating glucoamylases suitable for large scale protein purification procedures. Furthermore, different methods and uses related to glucoamylases according to the invention are disclosed, such as in a brewing process.

First claim

What is claimed is:

1. An isolated polypeptide having glucoamylase activity comprising an amino acid sequence having at least 90% sequence identity to an amino acid sequence selected from the group consisting of the mature polypeptide of SEQ ID NO: 3 and 6.

2. The polypeptide according to claim 1 , wherein the amino acid sequence comprises at least one or more amino acid residue(s) selected from the following groups:
a) an amino acid residue selected from the group consisting of A and P at a position corresponding to position 459 in SEQ ID NO: 3 or 6,
b) an amino acid residue selected from the group consisting of C and S at a position corresponding to position 473 in SEQ ID NO: 3 or 6,
c) an amino acid residue selected from the group consisting of A and R at a position corresponding to position 474 in SEQ ID NO: 3 or 6,
d) an amino acid residue selected from the group consisting of A and P at a position corresponding to position 475 in SEQ ID NO: 3 or 6,
e) an amino acid residue selected from the group consisting of T and Y at a position corresponding to position 476 in SEQ ID NO: 3 or 6,
f) an amino acid residue selected from the group consisting of P and G at a position corresponding to position 477 in SEQ ID NO: 3 or 6,
g) an amino acid residue selected from the group consisting of A and G at a position corresponding to position 479 in SEQ ID NO: 3 or 6 and/or
h) an amino acid residue selected from the group consisting of V and R at a position corresponding to position 480 in SEQ ID NO: 3 or 6.

3. The polypeptide according to claim 1 , comprising the mature polypeptide of SEQ ID NO:3 or 6.

4. The polypeptide according to claim 1 , wherein the polypeptide is inactivated by pasteurisation.

5. The polypeptide according to claim 1 , wherein the polypeptide is obtained by recombinant expression in a host cell.

6. A composition comprising the polypeptide of claim 1 .

7. The composition according to claim 6 , wherein the composition is selected from the group consisting of a starch hydrolyzing composition, a saccharifying composition, a detergent composition, an alcohol fermentation enzymatic composition, and an animal feed composition.

8. The composition according to claim 6 further comprising an additional enzyme.

9. The composition according to claim 8 , wherein the additional enzyme is selected from the group consisting of alpha-amylase, beta-amylase, peptidase (protease, proteinase, endopeptidase, exopeptidase), pullulanase, isoamylase, cellulase, endo-glucanase and related beta-glucan hydrolytic accessory enzymes, xylanase and xylanase accessory enzymes (for example, arabinofuranosidase, ferulic acid esterase, xylan acetyl esterase), acetolactate decarboxylase and glucoamylase, and combinations thereof.

10. The polypeptide according to claim 2 , wherein the polypeptide is inactivated by pasteurisation.

11. The polypeptide according to claim 3 , wherein the polypeptide is inactivated by pasteurisation.

Assignees

• Dupont Nutrition Biosci Aps

Inventors

• Cramer Jacob Flyvholm
• Fish Neville Marshall
• Degn Peter Edvard
• Nikolaev Igor
• Kruithof Paulien
• Van Solingen Piet
• Van Stigt Thans Sander
• Breneman Suzy
• Shetty Jayarama K
• Lee Sung Ho

Classifications

• C12C12/02

  Beer with low calorie content (C12C12/04 takes precedence) · CPC title

• Y02E50/17

  Cross-Sectional Technologies · mapped topic

• C12N9/2408 Primary

  acting on alpha -1,4-glucosidic bonds · CPC title

• C12Y302/01003

  Glucan 1,4-alpha-glucosidase (3.2.1.3), i.e. glucoamylase · CPC title

• C12C11/003

  Fermentation of beerwort · CPC title