Inventors:
William S. Adney - Golden CO, US
Michael E. Himmel - Littleton CO, US
Stephen R. Decker - Berthoud CO, US
Eric P. Knoshaug - Golden CO, US
Mark R. Nimlos - Golden CO, US
Michael F. Crowley - Lakewood CO, US
Tina Jeoh - Davis CA, US
Assignee:
Alliance for Sustainable Energy, LLC - Golden CO
International Classification:
C12N 9/26, C12N 1/20, C12N 15/00, C12P 21/06, C12P 21/04, C12Q 1/00, C12Q 1/68, C07H 21/04
US Classification:
435201, 435440, 435 691, 435 711, 4352523, 4353201, 435 61, 435 4, 536 232
Abstract:
Provided herein is an isolated Cel7A polypeptide comprising mutations in the catalytic domain of the polypeptide relative to the catalytic domain of a wild type Cel7A polypeptide, wherein the mutations reduce N-linked glycosylation of the isolated polypeptide relative to the wild type polypeptide. Also provided herein is an isolated Cel7A polypeptide comprising increased O-linked glycosylation of the linker domain relative to a linker domain of a wild type Cel7A polypeptide. The increased O-linked glycosylation is a result of the addition of and/or substitution of one or more serine and/or threonine residues to the linker domain relative to the linker domain of the wild type polypeptide. In some embodiments, the isolated Cel7A polypeptide comprising mutations in the catalytic domain of the polypeptide relative to the catalytic domain of a wild type Cel7A polypeptide further comprises increased O-linked glycosylation of the linker domain relative to a linker domain of a wild type Cel7A polypeptide. The mutations in the catalytic domain reduce N-linked glycosylation of the isolated polypeptide relative to the wild type polypeptide. The addition of and/or substitution of one or more serine and/or threonine residues to the linker domain relative to the linker domain of the wild type polypeptide increases O-linked glycosylation of the isolated polypeptide.