US 11,891,637 B2
GH61 polypeptide variants and polynucleotides encoding same
Janine Lin, Davis, CA (US); Doreen Bohan, Fairfield, CA (US); Michelle Maranta, Davis, CA (US); Leslie Beresford, Roseville, CA (US); Michael Lamsa, Woodland, CA (US); Matt Sweeney, Sacramento, CA (US); Mark David Wogulis, Davis, CA (US); Elizabeth Znameroski, Davis, CA (US); and Frank Winther Rasmussen, Roskilde (DK)
Assigned to Novozymes, Inc., Davis, CA (US)
Filed by Novozymes A/S, Bagsvaerd (DK)
Filed on Apr. 6, 2022, as Appl. No. 17/714,461.
Application 17/714,461 is a division of application No. 16/269,212, filed on Feb. 6, 2019, granted, now 11,345,905, issued on May 31, 2022.
Application 16/269,212 is a division of application No. 14/395,984, granted, now 10,227,579, issued on Mar. 12, 2019, previously published as PCT/US2013/038477, filed on Apr. 26, 2013.
Claims priority of provisional application 61/639,648, filed on Apr. 27, 2012.
Prior Publication US 2022/0307002 A1, Sep. 29, 2022
Int. Cl. C12N 9/24 (2006.01); C12P 19/02 (2006.01); C12P 19/14 (2006.01); C12N 9/42 (2006.01); C12N 9/02 (2006.01); D21C 5/00 (2006.01); D21H 17/00 (2006.01); C11D 3/386 (2006.01)
CPC C12N 9/2437 (2013.01) [C11D 3/38645 (2013.01); C12N 9/0071 (2013.01); C12N 9/2434 (2013.01); C12P 19/02 (2013.01); C12P 19/14 (2013.01); C12Y 302/01021 (2013.01); D21C 5/005 (2013.01); D21H 17/005 (2013.01); C12P 2203/00 (2013.01); C12Y 114/00 (2013.01); C12Y 302/01004 (2013.01)] 32 Claims
 
1. A variant of a parent GH61 polypeptide having cellulolytic enhancing activity, comprising a substitution at one or more positions corresponding to positions 138, 219, 26, 32, 34, 40, 41, 42, 47, 56, 72, 102, 123, 149, 152, 163, 164, 166, 169, 186, 200, 207, 213, 222, 234, 246, 249, and 250 of the mature polypeptide of SEQ ID NO: 30, wherein the substitution at position 138 is with Cys, Glu, Gly, Lys, Leu, or Met, the substitution at position 219 is with Glu, Met, Gln, or Cys, the substitution at position 26 is with Ile, the substitution at position 32 is with Glu or Ser, the substitution at position 34 is with Phe, the substitution at position 40 is with Ala, the substitution at position 41 is with Thr, the substitution at position 42 is with Ile, Glu, or Val, the substitution at position 47 is with Glu, Leu, or Arg, the substitution at position 56 is with Cys, Glu, or Thr, the substitution at position 72 is with Gln or Thr, the substitution at position 102 is with Lys or Pro, the substitution at position 123 is with Arg, the substitution at position 149 is with Ile, the substitution at position 152 is with Ser, the substitution at position 163 is with Glu, Phe, or Val, the substitution at position 164 is with Cys or Leu, the substitution at position 166 is with Leu, the substitution at position 169 is with Arg or Cys, the substitution at position 186 is with Phe, Lys, Thr, or Tyr, the substitution at position 200 is with Ile or Val, the substitution at position 207 is with Pro, the substitution at position 213 is with Glu, the substitution at position 222 is with Arg, the substitution at position 234 is with Gly or Lys, the substitution at position 246 is with Pro, the substitution at position 249 is with Gln, Arg, or Cys, and the substitution at position 250 is with Cys, wherein the variant has cellulolytic enhancing activity, wherein the variant has increased thermostability relative to the parent GH61 polypeptide, and wherein the parent GH61 polypeptide is selected from the group consisting of:
(a) a GH61 polypeptide having at least 90% sequence identity to the mature polypeptide of SEQ ID NO: 8 or 28; and
(b) a GH61 polypeptide encoded by a polynucleotide having at least 90% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 7 or 27;
wherein the variant has at least 90% but less than 100% sequence identity to the mature polypeptide of SEQ ID NO: 8 or 28.