Polynucleotides encoding polypeptides having alpha-amylase activity and methods of making the same

Patent metadata

Field	Value
Publication number	US-9994834-B2
Application number	US-201214345364-A
Country	US
Kind code	B2
Filing date	Sep 7, 2012
Priority date	Sep 9, 2011
Publication date	Jun 12, 2018
Grant date	Jun 12, 2018

Abstract

Provided are isolated polypeptides having alpha-amylase activity, catalytic domains, carbohydrate binding domains and polynucleotides encoding the polypeptides, catalytic domains or carbohydrate binding domains. Also provided are nucleic acid constructs, vectors and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides, catalytic domains or carbohydrate binding domains.

First claim

What is claimed is:

1. A non-natural nucleic acid construct or non-natural expression vector comprising a polynucleotide having at least 95% identity to the mature polypeptide coding sequence of SEQ ID NO:19 operably linked to one or more heterologous control sequences that direct the production of a polypeptide in an expression host, wherein the polynucleotide encodes a polypeptide having alpha-amylase activity.

2. A non-natural recombinant host cell comprising the nucleic acid construct or expression vector of claim 1 .

3. A method of producing a polypeptide having alpha-amylase activity, comprising:
(a) cultivating the host cell of claim 2 under conditions conducive for production of the polypeptide; and
(b) recovering the polypeptide.

4. A nucleic acid construct or expression vector comprising a gene encoding a protein, wherein the gene is operably linked to a polynucleotide encoding a signal peptide comprising or consisting of amino acids 1 to 16 of SEQ ID NO: 20, wherein the gene is foreign to the polynucleotide encoding the signal peptide, wherein the protein is selected from the group consisting of an aminopeptidase, amylase, carbohydrase, carboxypeptidase, catalase, cellulase, chitinase, cutinase, cyclodextrin glycosyltransferase, deoxyribonuclease, esterase, alpha-galactosidase, beta-galactosidase, glucoamylase, alpha-glucosidase, beta-glucosidase, invertase, laccase, lipase, mannosidase, mutanase, oxidase, pectinolytic enzyme, peroxidase, phytase, polyphenoloxidase, proteolytic enzyme, ribonuclease, transglutaminase and xylanase.

5. A method of producing a protein, comprising:
(a) cultivating a recombinant host cell transformed with the nucleic acid construct or expression vector of claim 4 , under conditions conducive for production of the protein; and
(b) recovering the protein.

6. A non-natural nucleic acid construct or non-natural expression vector comprising the mature polypeptide coding sequence of SEQ ID NO:19 operably linked to one or more heterologous control sequences that direct the production of a polypeptide in an expression host, wherein the polypeptide has at least 95% sequence identity to SEQ ID NO:20.

7. A non-natural nucleic acid construct or non-natural expression vector comprising a polynucleotide having at least 97% sequence identity to the mature polypeptide coding sequence of SEQ ID NO:19 operably linked to one or more heterologous control sequences that direct the production of a polypeptide in an expression host, wherein the polypeptide has at least 97% sequence identity to SEQ ID NO:20.

8. A non-natural expression vector comprising a polynucleotide having at least 99% sequence identity to the mature polypeptide coding sequence of SEQ ID NO:19 operably linked to one or more heterologous control sequences that direct the production of a polypeptide.

9. A composition comprising a polypeptide encoded by a polynucleotide having at least 95%, identity to the mature polypeptide coding sequence of SEQ NO: 19 operably linked to one or more heteroloqous control sequences and an enzyme selected from the group consisting of: a fungal alpha-amylase (EC 3.2.1.1), a beta-amylase (E.C. 3.2.1.2), a glucoamylase (E.C.3.2.1.3), a pullulanases (E.C. 3.2.1.41), a phytase (E.C.3.1.2.28) and a protease (E.C. 3.4.).

10. The nucleic acid construct or expression vector of claim 4 , wherein the gene is eukaryotic.

11. The nucleic acid construct or expression vector of claim 4 , wherein the gene is prokaryotic.

12. A recombinant host cell comprising the nucleic acid construct or expression vector of claim 4 .

13. The host cell of claim 12 , wherein the protein is native to the cell.

14. The host cell of claim 12 , wherein the protein is heterologous to the cell.

Assignees

• Sun tianqi
• Li Ming
• Duan Junxin
• Novozymes As

Inventors

• Sun tianqi
• Li Ming
• Duan Junxin

Classifications

• C12Y302/01001

  Alpha-amylase (3.2.1.1) · 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

• C12N9/242 Primary

  Fungal source · CPC title

• C12N9/2411

  Amylases · CPC title

• C12P19/02

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