Glycoside hydrolases having multiple hydrolase activities
AssigneesSandia Corporation, The Regents of the University of California
InventorsZhiwei Chen, Gregory D. Friedland, Swapnil R. Chhabra, Dylan C. Chivian, Blake A. Simmons
Life of this patent
Estimating the term…
Abstract
Glycoside hydrolases having at least two different hydrolytic activities are provided. In one embodiment, an isolated recombinant hydrolase having at least two activities selected from a group including asparagine derivatives, glutamine derivatives, and histidine derivatives is provided. Further, a method of generating free sugars from a mixture comprising asparagine derivatives, glutamine derivatives, and histidine derivatives is provided.
Claims21
An isolated recombinant glycoside hydrolase having at least two activities selected from the group consisting of cellulase activity, xylanase activity, and mannanase activity, wherein the glycoside hydrolase comprises the following amino acids (corresponding to positions in SEQ ID NO:1):
- asparagine (N), aspartic acid (D), glutamine (Q) or glutamic acid (E) at position 20, aspartic acid (D) or glutamic acid (E) at position 23, proline (P) at position 53, histidine (H) at position 95, histidine (H) at position 96 and aspartic acid (D) or glutamic acid (E) at position 287, wherein the recombinant glycoside hydrolase has an increased cellulase activity, xylanase activity, and/or mannanase activity compared to the glycoside hydrolase of SEQ ID NO:1.
A method of generating free sugars from a mixture comprising at least two polymeric substrates selected from the group consisting of cellulose, xylan and mannan, the method comprising contacting the mixture with a glycoside hydrolase under conditions such that the glycoside hydrolase hydrolyzes the at least two polymeric substrates, wherein the glycoside hydrolase comprises the following amino acids (corresponding to positions in SEQ ID NO:1):
- asparagine (N), aspartic acid (D), glutamine (Q) or glutamic acid (E) at position 20, aspartic acid (D) or glutamic acid (E) at position 23, proline (P) at position 53, histidine (H) at position 95, histidine (H) and position 96 and aspartic acid (D) or glutamic acid (E) at position 287, thereby generating free sugars.
A method of generating a polynucleotide encoding a mutated glycoside hydrolase with activity to hydrolyze at least two polymeric substrates selected from the group consisting of cellulose, xylan and mannan, the method comprising:
- introducing at least one nucleotide change to a coding region of a polynucleotide encoding a native glycoside hydrolase, resulting in at least one amino acid change in the glycoside hydrolase to generate a mutated polynucleotide encoding a mutated glycoside hydrolase, wherein the introducing changes at least one amino acid at positions 20, 23, 53, 95, 96, or 287 (corresponding to positions in SEQ ID NO:1) of the native glycoside hydrolase such that the mutated glycoside hydrolase comprises each of the following amino acids (corresponding to positions in SEQ ID NO:1):
- asparagine or aspartic acid at position 20, aspartic acid or glutamate at position 23, proline at position 53, histidine at position 95, histidine and position 96 and aspartate or glutamate at position 287, wherein the mutated glycoside hydrolase has an increased cellulase activity, xylanase activity, and/or mannanase activity compared to the native glycoside hydrolase.
Dependencies are inferred from the claim text.