US 7,960,148 B2 · Glucanases, nucleic acids encoding them and methods for making and using them

Abstract

The invention relates to polypeptides having glucanase, e.g., endoglucanase, mannanase, xylanase activity or a combination of these activities, and polynucleotides encoding them. In one aspect, the glucanase activity is an endoglucanase activity (e.g., endo-1,4-beta-D-glucan 4-glucano hydrolase activity) and comprises hydrolysis of 1,4-beta-D-glycosidic linkages in cellulose, cellulose derivatives (e.g., carboxy methyl cellulose and hydroxy ethyl cellulose) lichenin, beta-1,4 bonds in mixed beta-1,3 glucans, such as cereal beta-D-glucans or xyloglucans and other plant material containing cellulosic parts. In addition, methods of designing new enzymes and methods of use thereof are also provided. In alternative aspects, the new glucanases e.g., endoglucanases, mannanases, xylanases have increased activity and stability at increased pH and temperature.

Claims29

Dependent claims

An isolated, synthetic, or recombinant polypeptide comprising:

(i) an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or more or complete (100%) sequence identity to SEQ ID NO:38, or an enzymatically active fragment thereof, wherein the polypeptide or the enzymatically active fragment has a glucanase activity;
(ii) an amino acid sequence encoded by a nucleic acid having at least 95% sequence identity to SEQ ID NO.:
37, wherein the polypeptide has glucanase activity;
(iii) the amino acid sequence of (i), or (ii), lacking a signal sequence and/or a carbohydrate binding module
(iv) the amino acid sequence of (i), (ii), or (iii), further comprising a heterologous sequence;
or
(v) the amino acid sequence of (i), (ii), (iii) or (iv),
wherein the polypeptide comprises at least one glycosylation site;
or, wherein the polypeptide comprises at least one N-linked glycosylation site, or, wherein the polypeptide is glycosylated after being expressed in a P. pastoris or a S. pombe.

6 more elements

Dependencies are inferred from the claim text.

Glucanases, nucleic acids encoding them and methods for making and using them

Life of this patent

Glucanases, nucleic acids encoding them and methods for making and using them

Life of this patent

Abstract

Claims29