Glucoamylase variants and polynucleotides encoding same

Patent metadata

Field	Value
Publication number	US-9963690-B2
Application number	US-201414787422-A
Country	US
Kind code	B2
Filing date	Apr 29, 2014
Priority date	Apr 30, 2013
Publication date	May 8, 2018
Grant date	May 8, 2018

Abstract

The present invention relates to glucoamylase variants. 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 one or more positions corresponding to positions 45, 46, 61, 86, 119, 120, 318, 348 of the polypeptide of SEQ ID NO: 3 wherein the variant has glucoamylase activity and wherein the variant has at least 80%, but less than 100% sequence identity to the polypeptide of SEQ ID NO: 3.

2. The variant according to claim 1 , wherein the number of substitutions is 1-20.

3. The variant of claim 1 , wherein the variant comprises or consists of one or more substitutions selected from the group consisting of T45K, S46H, V61A, V61T, S86C, T119W, D120G, M318W, N348T and wherein the variant has glucoamylase activity and wherein the variant has at least 80%, but less than 100% sequence identity to the polypeptide of SEQ ID NO: 3.

4. The variant of claim 1 , wherein the variant comprises at least one of the following substitutions or combinations of substitutions:
T119W;
V61A;
D120G;
S86C+T119W;
T45K+T119W;
T119W+D120G;
T45K+D120G;
V61T+T119W;
D120G+M318W;
T45K+S46H;
T119W +D120G+N348T;
T119W+D120G+M318W;
S86C+T119W+D120G;
S46H+T119W+D120G;
T45K+T119W+D120G and wherein the variant has glucoamylase activity and wherein the variant has at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100% sequence identity to the polypeptide of SEQ ID NO: 3.

5. The variant of claim 1 , which has an improved property relative to the parent, wherein the improved property is increased thermostability.

6. A composition comprising the polypeptide of claim 1 .

7. The composition according to claim 6 , comprising an alpha-amylase and a polypeptide of claim 1 .

8. A process of producing a fermentation product from starch-containing material comprising the steps of: (a) liquefying starch-containing material in the presence of an alpha amylase; (b) saccharifying the liquefied material; and (c) fermenting with a fermenting organism; wherein step (a) and/or step (b) is carried out using at least a glucoamylase variant of claim 1 .

9. A process of producing a syrup product from starch-containing material, comprising the step of: (a) liquefying starch-containing material in the presence of an alpha amylase; (b) saccharifying the liquefied material in the presence of a variant glucoamylase of claim 1 .

10. An isolated polynucleotide encoding the variant of claim 1 .

11. A nucleic acid construct comprising the polynucleotide of claim 10 .

12. An expression vector comprising the polynucleotide of claim 10 .

13. A host cell comprising the polynucleotide of claim 10 .

14. A method of producing a glucoamylase variant, comprising: cultivating the host cell of claim 13 under conditions suitable for expression of the variant; and recovering the variant glucoamylase.

15. The variant of claim 1 , having at least at least 85% sequence identity to the polypeptide of SEQ ID NO: 3.

16. The variant of claim 1 , having at least at least 90% sequence identity to the polypeptide of SEQ ID NO: 3.

17. The variant of claim 1 , having at least at least 95% sequence identity to the polypeptide of SEQ ID NO: 3.

18. The variant of claim 1 , having at least at least 97% sequence identity to the polypeptide of SEQ ID NO: 3.

19. The variant of claim 1 , having at least at least 98% sequence identity to the polypeptide of SEQ ID NO: 3.

20. The variant of claim 1 , having at least at least 99% sequence identity to the polypeptide of SEQ ID NO: 3.

Assignees

• Novozymes As

Inventors

• Vind Jesper
• Mepadam Vasu Santhosh
• Saikia Rakhi
• Basu Aditya

Classifications

• C12P19/02

  Monosaccharides (2-ketogulonic acid C12P7/60) · CPC title

• C12N9/2408

  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

• C12N9/2428 Primary

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

• C12P19/20

  produced by the action of an exo-1,4 alpha-glucosidase, e.g. dextrose · CPC title