US 12,391,934 B2
Stabilized cellulase variants
Svend Gunnar Kaasgaard, Svoklunde (DK); Frank Winther Rasmussen, Roskilde (DK); Roland Alexander Pache, Valby (DK); Lars Anderson, Malmoe (SE); Mette Louise Dissing Overgaard, Copenhagen (DK); Thomas Agersten Poulsen, Ballerup (DK); Dorte Marie Koefoed Klitgaard, Copenhagen (DK); Christian Isak Joergensen, Bagsvaerd (DK); and Lars Giger, Valby (DK)
Assigned to Novozymes A/S, Bagsvaerd (DK)
Filed by Novozymes A/S, Bagsvaerd (DK)
Filed on Jan. 11, 2023, as Appl. No. 18/152,992.
Application 18/152,992 is a division of application No. 17/041,588, granted, now 11,566,239, previously published as PCT/EP2019/059462, filed on Apr. 12, 2019.
Claims priority of application No. 18168286 (EP), filed on Apr. 19, 2018.
Prior Publication US 2023/0295591 A1, Sep. 21, 2023
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 9/42 (2006.01); C11D 3/386 (2006.01)
CPC C12N 9/2437 (2013.01) [C11D 3/38645 (2013.01); C12Y 302/01004 (2013.01)] 20 Claims
 
1. A variant having endoglucanase activity, where the variant has at least 85% sequence identity to the endoglucanase having SEQ ID NO: 4, comprising two or more substitutions in positions corresponding to following positions in SEQ ID NO: 1: 25, 137, 152, or 169, using SEQ ID NO: 1 for numbering, and where
the substitution at position 25 is with R (arqinine), N (asparaqine), D (aspartic acid), C (cysteine), Q (qlutamine), E (qlutamic acid), G (qlycine), H (histidine), I (isoleucine), L (leucine), K (lysine), M (methionine), F (phenylalanine), P (proline), S (serine), T (threonine), W (tryptophan), Y (tyrosine), or V (valine);
the substitution at position 137 is with A (alanine), R (arginine), N (asparagine), D (aspartic acid), C (cysteine), Q (qlutamine), E (qlutamic acid), G (qlycine), H (histidine), I (isoleucine), L (leucine), K (lysine), M (methionine), F (phenylalanine), P (proline), T (threonine), W (tryptophan), Y (tyrosine), and V (valine);
the substitution at position 152 is with A (alanine), R (arginine), N (asparagine), D (aspartic acid), C (cysteine), Q (qlutamine), E (qlutamic acid), G (qlycine), H (histidine), I (isoleucine), L (leucine), K (lysine), M (methionine), F (phenylalanine), P (proline), T (threonine), W (tryptophan), Y (tyrosine), and V (valine);
the substitution at position 169 is with A (alanine), R (arqinine), N (asparaqine), D (aspartic acid), C (cysteine), E (qlutamic acid), G (qlycine), H (histidine), I (isoleucine), L (leucine), K (lysine), M (methionine), F (phenylalanine), P (proline), S (serine), T (threonine), W (tryptophan), Y (tyrosine), and V (valine); and
the variant has improved stability in an aqueous composition comprising a protease in comparison with the parent endoglucanase.