Alicyclobacillus variants and polynucleotides encoding same
AssigneeNovozymes A/S
InventorsCarsten Andersen, Iben Damager, Astrid Munch, Tine Hoff
Life of this patent
Estimating the term…
Abstract
The present invention relates to alpha-amylase variants. The present invention also relates to polynucleotides encoding the variants; nucleic acid constructs, vectors, and host cells comprising the polynucleotides; and methods of using the variants.
Claims32
An alpha-amylase variant of a parent alpha-amylase, comprising a substitution at one or more positions corresponding to positions 109, 51, 201, 269, 294, 297, 298, 193, and 314 of the polypeptide according to SEQ ID NO:3, wherein said variant has alpha-amylase activity, and wherein said parent alpha-amylase has at least 89%, such as at least 90%, such as at least 91%, such as at least 92%, such as at least 93%, such as at least 94%, such as at least 95%, sequence identity to the polypeptide of SEQ ID NOs:
- 3 or 13.
A method for obtaining an alpha-amylase variant, comprising introducing into a parent alpha-amylase having at least 89% sequence identity to the polypeptide of SEQ ID NOs:
- 3 or 13 a substitution at one or more positions said substitutions corresponding to positions 109, 51, 201, 269, 294, 297, 298, 193, and 314 of SEQ ID NO:3, wherein the variant has at least 67%, such as at least 70%, such as at least 75%, such as at least 80%, such as at least 85%, such as at least 90%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 99%, but less than 100% sequence identity with the amino acid sequence of SEQ ID NOs:
- 3 or 13, wherein the variant has alpha-amylase activity;
- and recovering said variant.
A method of improving the detergent stability of a parent alpha-amylase having the amino acid sequence of SEQ ID NOs:
- 3 or 13, or having at least 89% sequence identity thereto, said method comprising the steps of:
- a) a substitution at one or more positions said substitutions corresponding to positions 109, 51, 201, 269, 294, 297, 298, 193, and 314 of SEQ ID NO:3, wherein the variant has at least 67%, such as at least 70%, such as at least 75%, such as at least 80%, such as at least 85%, such as at least 90%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 99%, but less than 100% sequence identity with the amino acid sequence of SEQ ID NOs:
- 3 or 13, wherein the variant has alpha-amylase activity;
- and
- b) introducing into the parent alpha-amylase one or more of the following substitutions;
- 105L,I,F+206Y;
- 105L,I+206Y+217I;
- 105F+206Y+208Y+217V+246V;
- 105L+206F;
- 1051+206Y+208Y+217I+246V;
- 195F+2135+214T;
- 195F+206Y+208Y+213T+214T+217M,V;
- 195F+206Y+208F,L+213T+214T+217V;
- 195F+206Y+2135+214T;
- 195F+206Y+208Y+213S+214T+217M;
- 195F+206Y+208F+213T+214T+217M;
- 195F+206Y+208Y+213T+214T+217Q;
- 195F+206Y+213G+214T;
- 195F+206Y+213S;
- 195F+206Y+208Y+213T+214T+217M;
- 195F+213S;
- 195F+206Y+208L+213T+214T+217M;
- 195F+213G+214T;
- 206Y,F+208Y+217Q;
- 206Y+208Y+217I;
- 206F+208Y+217M;
- 206Y+208Y;
- 206Y+217M;
- 206Y+208Y+213A+217M;
- 206Y+208Y+217V+246V;
- 206Y+213G;
- 206Y+208F+217V;
- 206N+208Y+217M;
- 206F+208Y+217V;
- 206Y+246V;
- 206Y+217I,V;
- 206F+208F+217I;
- 206Y+208L+213S;
- 206F+217I;
- 206Y+217I+246I;
- 206L+217V;
- 206Y+208F+217H;
- 206L+208F+217I;
- 206L+217V+246L;
- 206F+246V;
- 208Y+213S+217M;
- 208Y+213A+217Q;
- 631+206Y;
- 63I+206Y+241V;
- 63V+206Y;
- 63V+105L+206Y;
- 63V+206Y+2171;
- 63V+105F+206Y+208F+217I;
- 63V+206Y+246V;
- 63V+206F;
- 63V+206L+217V;
- 63V+105F+206Y;
- 63V+206Y+241V+246L;
- 195F+206Y+208Y+214T+217V;
- 186E+195F+206Y;
- 195F+206Y+208Y+213T+217V;
- 186E+195F+202T+206Y+209S;
- 63I+195F+206Y+210S;
- 195F+206Y+213P+214T;
- 195F+206Y+208Y+213T+214T+217I;
- 186E+195F+206Y+210S;
- 195F+213P;
- 186E+195F+202T+206Y+210S;
- 195F+206H;
- 195F+208Y+213T+214T+217V;
- 206Y+208Y+213T+214T+217V;
- 195F+206Y+217V;
- 195F+206Y+208Y+213S+214T;
- 195F+206Y+208Y;
- 195F+213I+214P;
- 195F+206Y+208Y+213T+214T;
- 195F+206Y;
- 206Y+213S;
- 182P+186E;
- 182S+186E;
- 182V+186K;
- 179L+186H+190P;
- 179L+186K,R,S+190P;
- 179L+190P;
- 179L+182C+186K+190P;
- 179L+182P+186S,V+190P;
- 179L+182S+186Q+190P;
- 173F+174Q;
- 173Y+174S;
- 172K+173Y+174E;
- 193A,D,N,S+195F;
- 213A+214Q;
- 213P+214L;
- 213S+214R;
- 48V+60V;
- 213G+214T;
- 213I+214P;
- 213N+214I;
- 213N+214Q, and 213P,S+214T, wherein numbering is according to SEQ ID NO:11,
- wherein the variant has at least 67%, such as at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, but less than 100% sequence identity with the polypeptide of SEQ ID NOs:
- 3 or 13, and wherein the variant has alpha-amylase activity and improved detergent stability and/or improved performance compared to the parent alpha-amylase.
(canceled)
Dependencies are inferred from the claim text.