# US 11,898,177 B2 · Variant maltohexaose-forming alpha-amylase variants

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- Number: US 11,898,177 B2
- Document ID: US11898177B2
- Status: Granted
- Kind: B2
- Application number: 17/373,307
- Filed: 2021-07-12
- Published: 2022-01-06
- Granted: 2024-02-13
- Earliest claimed: 2011-10-28
- Assignees: Danisco US Inc.
- Applicants: Danisco US Inc.
- Inventors: Luis G Cascao-Pereira; David A Estell; Marc Anton Bernhard Kolkman; Harm Mulder
- Web page: https://patentgrep.com/patent/US11898177B2

## CPC

- C12N 9/2417: {from microbiological source}
- A21D 8/042: {with enzymes}
- C11D 3/386: Preparations containing enzymes {, e.g. protease or amylase}
- C12P 7/06: Ethanol, i.e. non-beverage
- C12P 19/14: produced by the action of a carbohydrase {(EC 3.2.x)}, e.g. by alpha-amylase {, e.g. by cellulase, hemicellulase}
- C12Y 302/01001: Alpha-amylase (3.2.1.1)
- D06L 1/14: De-sizing
- D06L 4/40: using enzymes
- Y02E 50/10: Biofuels, e.g. bio-diesel
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- A21D: TREATMENT OF FLOUR OR DOUGH FOR BAKING, e.g. BY ADDITION OF MATERIALS; BAKING; BAKERY PRODUCTS (preservation of flour or dough before baking A23B40/00; preservation of bakery products A23B45/00)
- C11D: DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
- C12P: FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {(brewing of beer C12C; producing vinegar C12J; producing specific peptides or proteins C07K; producing enzymes C12N9/00; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; measuring or testing processes involving enzymes or microorganisms C12Q; measuring or testing processes involving nucleic acid amplification reactions C12Q1/6844; fermentation processes to form a food composition, A21 or A23; compounds in general, see the relevant compound class, e.g. C01, C07)}
- C12Y: ENZYMES
- D06L: DRY-CLEANING, WASHING OR BLEACHING FIBRES, FILAMENTS, THREADS, YARNS, FABRICS, FEATHERS OR MADE-UP FIBROUS GOODS; BLEACHING LEATHER OR FURS
- Y02E: REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION

## Abstract

Disclosed are compositions and methods relating to variant maltohexaose-forming alpha-amylases. The variant alpha-amylases are useful, for example, for starch liquefaction and saccharification, for cleaning starchy stains in laundry, dishwashing, and other applications, for textile processing (e.g., desizing), in animal feed for improving digestibility, and for baking and brewing.

## Claims

### Claim 1

A variant α-amylase polypeptide derived from a parental α-amylase polypeptide, comprising at least two combinable mutations at productive amino acid positions; wherein the mutations are at position H1 and E391, wherein the variant has at least 90% amino acid sequence identity with SEQ ID NO: 3.

### Claim 2 (depends on 1)

The variant amylase of claim 1, wherein the mutations are H1A and E391A.

### Claim 3 (depends on 1)

The variant amylase of claim 1, further comprising a plurality of combinable mutations.

### Claim 4 (depends on 1)

The variant amylase of claim 1, further comprising a deletion corresponding to one or more residue selected from the group consisting of Arg-181, Gly-182, His-183, and Gly-184, using SEQ ID NO: 3 for numbering.

### Claim 5 (depends on 1)

The variant amylase of claim 1, further comprising deletions corresponding to residues Arg-181 and Gly-182, using SEQ ID NO: 3 for numbering.

### Claim 6 (depends on 1)

A composition comprising the variant amylase of claim 1.

### Claim 7 (depends on 6)

The composition of claim 6, wherein the composition is effective for removing starchy stains from laundry, dishes, or textiles.

### Claim 8 (depends on 6)

The composition of claim 6, further comprising a surfactant.