Thermostable alpha-amylases

Patent metadata

Field	Value
Publication number	US-9394533-B2
Application number	US-201414231062-A
Country	US
Kind code	B2
Filing date	Mar 31, 2014
Priority date	Dec 17, 2002
Publication date	Jul 19, 2016
Grant date	Jul 19, 2016

Abstract

The present invention relates to an isolated polynucleotide comprising an open reading frame encoding a polypeptide having alpha-amylase activity, the polypeptide selected from the group consisting of:
a) a polypeptide comprising an amino acid sequence which has at least 70% identity with amino acids 22 to 450 of SEQ ID NO: 4; b) a polypeptide comprising an amino acid sequence which has at least 70% identity with the polypeptide encoded by the amylase encoding part of the polynucleotide inserted into a plasmid present in the E. coli host deposited under the Budapest Treaty with DSMZ under accession number DSM 15334; c) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence which has at least 70% identity with the sequence shown from position 68 to 1417 in SEQ ID NO: 3; and d) a fragment of (a), (b) or (c) that has alpha-amylase activity.

First claim

The invention claimed is:

1. A recombinant expression vector comprising a nucleic acid construct comprising a polynucleotide operably linked to one or more control sequences that direct the production of a polypeptide in a suitable host cell; wherein the polynucleotide comprises an open reading frame encoding a polypeptide having alpha-amylase activity, the polypeptide selected from the group consisting of:
a) a polypeptide comprising an amino acid sequence which has at least 90% sequence identity with amino acids 22 to 450 of SEQ ID NO: 4;
b) a polypeptide comprising an amino acid sequence which has at least 90% sequence identity with the polypeptide encoded by the amylase encoding part of the polynucleotide inserted into a plasmid present in the E. coli host deposited under the Budapest Treaty with DSMZ under accession number DSM 15334;
c) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence which has at least 90% identity with the sequence shown from position 68 to 1417 in SEQ ID NO: 3; and
d) a fragment of (a), (b) or (c) that has alpha-amylase activity.

2. A recombinant host cell comprising a nucleic acid construct comprising a polynucleotide operably linked to one or more control sequences that direct the production of a polypeptide in a suitable host cell; wherein the polynucleotide comprises an open reading frame encoding a polypeptide having alpha-amylase activity, the polypeptide selected from the group consisting of:
a) a polypeptide comprising an amino acid sequence which has at least 90% sequence identity with amino acids 22 to 450 of SEQ ID NO: 4;
b) a polypeptide comprising an amino acid sequence which has at least 90% sequence identity with the polypeptide encoded by the amylase encoding part of the polynucleotide inserted into a plasmid present in the E. coli host deposited under the Budapest Treaty with DSMZ under accession number DSM 15334;
c) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence which has at least 90% identity with the sequence shown from position 68 to 1417 in SEQ ID NO: 3; and
d) a fragment of (a), (b) or (c) that has alpha-amylase activity.

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

4. A method of producing an enzymatically modified starch derivative, wherein a polypeptide having alpha-amylase activity produced according to a method of claim 3 is used for saccharifying starch.

5. A method for producing a polypeptide having alpha-amylase activity, the method comprising:
(a) cultivating a recombinant host cell comprising a nucleic acid construct comprising a polynucleotide operably linked to one or more control sequences that direct the production of the polypeptide in a suitable host cell; wherein the polynucleotide comprises an open reading frame encoding a polypeptide having alpha-amylase activity, the polypeptide comprising an amino acid sequence which has at least 95% sequence identity with amino acids 22 to 450 of SEQ ID NO: 4 under conditions conducive for production of the polypeptide; and
(b) recovering the polypeptide.

6. The method of claim 5 , wherein the polypeptide comprises an amino acid sequence which has at least 99% sequence identity with amino acids 22 to 450 of SEQ ID NO: 4.

7. A method of making alcohol comprising:
producing a polypeptide having alpha-amylase activity, the method comprising:
(a) cultivating a recombinant host cell comprising a nucleic acid construct comprising a polynucleotide operably linked to one or more control sequences that direct the production of the polypeptide in a suitable host cell; wherein the polynucleotide comprises an open reading frame encoding a polypeptide having alpha-amylase activity, the polypeptide comprising an amino acid sequence which has at least 95% sequence identity with amino acids 22 to 450 of SEQ ID NO: 4 under conditions conducive for production of the polypeptide;
(b) recovering the polypeptide;
(c) saccharrifying a starch material by contacting the starch material with the polypeptide to form a sugar; and
(d) fermenting the sugar to form alcohol.

Assignees

• Novozymes As

Inventors

• Tang Lan
• Wu Wenping
• Duan Junxin
• Johannesen Pia Francke

Classifications

• C12N9/2414

  Alpha-amylase (3.2.1.1.) · CPC title

• C12Y302/01001

  Alpha-amylase (3.2.1.1) · CPC title

• C12C11/003

  Fermentation of beerwort · CPC title

• A21D8/042

  with enzymes · 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