# US 8,465,957 B2 · Alpha-amylase variants

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- Number: US 8,465,957 B2
- Document ID: US8465957B2
- Status: Granted
- Kind: B2
- Application number: 13/478,512
- Filed: 2012-05-23
- Published: 2012-10-04
- Granted: 2013-06-18
- Earliest claimed: 1998-11-16
- Assignees: Novozymes A/S
- Applicants: Allan Svendsen; Soren Kjaerulff; Henrik Bisgaard-Frantzen; Carsten Andersen
- Inventors: Allan Svendsen; Soren Kjaerulff; Henrik Bisgaard-Frantzen; Carsten Andersen
- Web page: https://patentgrep.com/patent/US8465957B2

## CPC

- C12N 9/2414: {Alpha-amylase (3.2.1.1.)}
- C07H 21/04: with deoxyribosyl as saccharide radical
- C11D 3/38609: {Protease or amylase in solid compositions only}
- C11D 3/38618: {Protease or amylase in liquid compositions only}
- C12N 9/2417: {from microbiological source}
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C07H: SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS (derivatives of aldonic or saccharic acids C07C, C07D; aldonic acids, saccharic acids C07C59/105, C07C59/285; cyanohydrins C07C255/16; glycals C07D; compounds of unknown constitution C07G; polysaccharides, derivatives thereof C08B; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; sugar industry C13)
- C11D: DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL

## Abstract

The invention relates to a variant of a parent Termamyl-like alpha-amylase, comprising mutations in two, three, four, five or six regions/positions. The variants have increased stability at high temperatures (relative to the parent). The invention also relates to a DNA construct comprising a DNA sequence encoding an alpha-amylase variant of the invention, a recombinant expression vector which carries a DNA construct of the invention, a cell which is transformed with a DNA construct of the invention, the use of an alpha-amylase variant of the invention for washing and/or dishwashing, textile desizing, starch liquefaction, a detergent additive comprising an alpha-amylase variant of the invention, a manual or automatic dishwashing detergent composition comprising an alpha-amylase variant of the invention, a method for generating a variant of a parent Termamyl-like alpha-amylase, which variant exhibits increased.

## Claims

### Claim 1

A variant alpha-amylase, said variant has an amino acid sequence which is at least 90% identical to SEQ ID NO: 3, has alpha-amylase activity and comprises an insertion at a position corresponding to position 376 of SEQ ID NO:4.

### Claim 2 (depends on 1)

The variant of claim 1, wherein the variant has an amino acid sequence which is at least 95% identical to SEQ ID NO: 3.

### Claim 3 (depends on 1)

The variant of claim 1, wherein the variant has an amino acid sequence which is at least 97% identical to SEQ ID NO: 3.

### Claim 4 (depends on 1)

The variant of claim 1, wherein the variant has an amino acid sequence which is at least 99% identical to SEQ ID NO: 3.

### Claim 5 (depends on 1)

A detergent composition comprising the variant alpha-amylase of claim 1 and surfactant.

### Claim 6 (depends on 5)

A detergent composition of claim 5, further comprising an enzyme selected from the group consisting of a protease, a lipase, a peroxidase, another amylolytic enzyme, a cellulase, and combinations of any of the foregoing.