Endoglucanase 1b (eg1b) variants
AssigneeCodexis, Inc.
InventorsEe Lui Ang, Ellen D. Eberhard, Xiyun Zhang, Wei Zhang, Jing Tian, Derek J. Smith, Vesna Mitchell
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Abstract
The present invention provides endoglucanase 1b (EG1b) variants suitable for use in saccharification reactions. The present application further provides genetically modified fungal organisms that produce EG1b variants, as well as enzyme mixtures exhibiting enhanced hydrolysis of cellulosic material to fermentable sugars, enzyme mixtures produced by the genetically modified fungal organisms, and methods for producing fermentable sugars from cellulose using such enzyme mixtures.
Claims51
A variant endoglucanase or biologically active variant endoglucanase fragment comprising (a) an amino acid sequence comprising at least about 70%, at least about 75%, at least about 80%, at least about 81%, at least 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to a wild-type endoglucanase (SEQ ID NO:3);
- and (b) an amino acid substitution from one or more of the amino acid residues selected from 5, 6, 7, 8, 9, 10, 11, 12, 13, 14,15, 16, 17, 18, 19, 20, 21, 22, 34, 38, 39, 43, 49, 63, 66, 75, 85, 126, 137, 143, 201, 218, 228, 241, 269, 274, 282, 328, 342, 342, 350, 397, 398, 399, 400, 401, 403, 404, 405, 407, 408, 409, 411, 412, 413, 414, 415, 417, 418, 419, 420, 425, 428, 433, 434, 435, 438, 439, 440, 443, 444, 446, 457, and 464, wherein the residues are numbered with reference to SEQ ID NO:2.
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A recombinant nucleic acid construct comprising at least one polynucleotide sequence encoding at least one variant endoglucanase, wherein the polynucleotide is selected from:
- (a) a polynucleotide that encodes a polypeptide comprising an amino acid sequence comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to SEQ ID NOS:5, 7 and/or 9, wherein the amino acid sequence comprises at least one substitution and/or substitution set provided herein;
- (b) a polynucleotide that hybridizes under stringent hybridization conditions to at least a fragment of a polynucleotide that encodes a polypeptide comprising the amino acid sequence of SEQ ID NOS:
- 1, 4, 6, and/or 8, and wherein said amino acid sequence comprises at least one substitution and/or at least one substitution set provided herein;
- and/or
- (c) a polynucleotide that hybridizes under stringent hybridization conditions to the complement of at least a fragment of a polynucleotide that encodes a polypeptide comprising the amino acid sequence of SEQ ID NOS:
- 1, 4, 6, and/or 8, and wherein said amino acid sequence comprises at least one substitution and/or at least one substitution set provided herein.
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Dependencies are inferred from the claim text.