Variant maltohexaose-forming alpha-amylase variants
AssigneeDanisco US Inc.
InventorsLuis G. CASCAO-PEREIRA, David A. ESTELL, Marc Anton Bernhard KOLKMAN, Harm MULDER
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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.
Claims76
A variant α-amylase polypeptide derived from a parental α-amylase polypeptide, comprising at least one combinable mutation at a productive amino acid position;
- wherein:
- (i) the combinable mutation is the substitution of an amino acid residue present in the parental α-amylase with a different amino acid residue, which improves at least one desirable property of the variant α-amylase compared to the parental α-amylase, while not significantly decreasing either expression, activity, or stability of the variant α-amylase, compared to the parental α-amylase,
- (ii) the productive position is an amino acid position that can be substituted with a plurality of different amino acid residues, each of which substitutions result in a variant α-amylase that meets the requirements of (i), and
- (iii) the combinable mutation corresponds to a mutation listed in Lists A, B, C, or D, or in Table C or D, which use SEQ ID NO:
- 3 for numbering.
The variant amylase of any of the preceding claims, wherein the combinable mutation has a suitability score of +++, ++++, or +++++, referring to Table B.
The variant amylase of any of the preceding claims, wherein the combinable mutation has a suitability score of ++++, or +++++, referring to Table B.
The variant amylase of any of the preceding claims, wherein the combinable mutation has a suitability score of +++++, referring to Table B.
The variant amylase of any of the preceding claims, wherein the combinable mutation has a productivity score of 1 or 2.
The variant amylase of any of the preceding claims, having a plurality of combinable mutations.
The variant amylase of any of the preceding claims, further comprising a deletion corresponding to a residue selected from the group consisting of Arg-181, Gly-182, His-183, and Gly-184, using SEQ ID NO:
- 3 for numbering.
The variant amylase of any of the preceding claims, further comprising deletions corresponding to residues Arg-181 and Gly-182, using SEQ ID NO:
- 3 for numbering.
The variant amylase of any of the preceding claims, wherein the parental α-amylase or the variant α-amylase has at least 60% amino acid sequence identity to the amino acid sequence of SEQ ID NO:
- 3 or SEQ ID NO:
- 4, or wherein the parental α-amylase or the variant α-amylase is encoded by a nucleic acid that hybridizes under stringent conditions to the nucleic acid of SEQ ID NO:
- 1 or SEQ ID NO:
- 5.
The variant amylase of any of the preceding claims, wherein the parental α-amylase or the variant α-amylase has at least 70% amino acid sequence identity to the amino acid sequence of SEQ ID NO:
- 3 or SEQ ID NO:
- 4, or wherein the parental α-amylase or the variant α-amylase is encoded by a nucleic acid that hybridizes under stringent conditions to the nucleic acid of SEQ ID NO:
- 1 or SEQ ID NO:
- 5.
The variant amylase of any of the preceding claims, wherein the parental α-amylase or the variant α-amylase has at least 80% amino acid sequence identity to the amino acid sequence of SEQ ID NO:
- 3 or SEQ ID NO:
- 4, or wherein the parental α-amylase or the variant α-amylase is encoded by a nucleic acid that hybridizes under stringent conditions to the nucleic acid of SEQ ID NO:
- 1 or SEQ ID NO:
- 5.
The variant amylase of any of the preceding claims, wherein the parental α-amylase or the variant α-amylase has at least 90% amino acid sequence identity to the amino acid sequence of SEQ ID NO:
- 3 or SEQ ID NO:
- 4, or wherein the parental α-amylase or the variant α-amylase is encoded by a nucleic acid that hybridizes under stringent conditions to the nucleic acid of SEQ ID NO:
- 1 or SEQ ID NO:
- 5.
The method of claim 46, wherein the method further comprises:
- (a) preparing a mash;
- (b) filtering the mash to obtain a wort;
- and
- (c) fermenting the wort to obtain a fermented beverage,
- wherein the variant amylase of any one of claims 1-16 and 74-80 is added to:
- (i) the mash of step (a) and/or
- (ii) the wort of step (b) and/or
- (iii) the wort of step (c).
Dependencies are inferred from the claim text.