Glucoamylase variants and polynucleotides encoding same

Patent metadata

Field	Value
Publication number	US-9534239-B2
Application number	US-201214343278-A
Country	US
Kind code	B2
Filing date	Sep 5, 2012
Priority date	Sep 6, 2011
Publication date	Jan 3, 2017
Grant date	Jan 3, 2017

Abstract

The present invention relates to glucoamylase variants having reduced sensitivity to protease nicking. The present invention also relates to polynucleotides encoding the variants; nucleic acid constructs, vectors, and host cells comprising the polynucleotides; and methods of using the variants.

First claim

The invention claimed is:

1. A glucoamylase variant, comprising a substitution at least at a position corresponding to position 79 of the mature polypeptide of SEQ ID NO: 2, wherein the variant has glucoamylase activity.

2. The glucoamylase variant of claim 1 , selected from the group consisting of:
a) a polypeptide having at least 90% sequence identity to the mature polypeptide of SEQ ID NO: 2; and
b) a polypeptide encoded by a polynucleotide that hybridizes under high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 1, or (ii) the full-length complement of (i), wherein the high stringency conditions include hybridization in 50% formamide, 1 M NaCl, 1% SDS at 37° C.

3. The glucoamylase variant of claim 1 , wherein the variant is a polypeptide having at least 95% sequence identity to the polypeptide of SEQ ID NO: 2.

4. The glucoamylase variant of claim 1 , wherein the number of substitutions is 1-20, 1-10 or 1-5.

5. The glucoamylase variant of claim 1 , wherein the variant comprises a substitution selected from K79V, K79A, K79G, K79I, K79L, K79S, K79T.

6. The glucoamylase variant of claim 1 , wherein the variant comprises substitution K79V.

7. A variant glucoamylase catalytic domain comprising a substitution at least at a position corresponding to position 79 of the mature polypeptide of SEQ ID NO: 2, wherein the variant has glucoamylase activity.

8. The variant glucoamylase catalytic domain of claim 7 selected from the group consisting of:
(a) a catalytic domain having at least 95% sequence identity to amino acids 30 to 494 of SEQ ID NO: 2; and wherein the catalytic domain has glucoamylase activity.

9. A composition comprising the glucoamylase variant of claim 1 .

Assignees

• Matsui Tomoko
• Clark Suzanne
• Novozymes As

Inventors

• Matsui Tomoko
• Clark Suzanne

Classifications

• C12C5/004 Primary

  Enzymes · CPC title

• Y02P20/52

  using catalysts, e.g. selective catalysts · CPC title

• C13K1/06

  obtained by saccharification of starch or raw materials containing starch · CPC title

• C12P7/14 Primary

  Multiple stages of fermentation; Multiple types of microorganisms or re-use of microorganisms · CPC title

• C12Y302/01003

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