Enhanced cellulose degradation

Patent metadata

Field	Value
Publication number	US-2016168609-A1
Application number	US-201514941492-A
Country	US
Kind code	A1
Filing date	Nov 13, 2015
Priority date	Apr 4, 2011
Publication date	Jun 16, 2016
Grant date	—

Abstract

The present disclosure provides compositions and methods related to the degradation of cellulose and cellulose-containing materials. CDH-heme domain polypeptides and GH61 polypeptides and related polynucleotides and compositions are provided herein. Additionally, methods related to CDH-heme domain polypeptides, GH61 polypeptides, and related polynucleotides and compositions, are provided herein

First claim

1 - 18 . (canceled)

19 . A method of degrading cellulose, the method comprising contacting the cellulose with:
one or more cellulases, a recombinant GH61 polypeptide; and a recombinant CDH-heme domain polypeptide comprising a cellulose binding module (CBM),
wherein the contact occurs in a reaction mixture, and
wherein the contact occurs for a time sufficient to yield degraded cellulose.

20 - 27 : (canceled)

28 . The method of claim 19 , wherein at least 50% of the GH61 polypeptides are bound to a copper atom.

29 . The method of claim 19 , wherein at least 90% of the GH61 polypeptides are bound to a copper atom.

30 - 31 : (canceled)

32 . The method of claim 19 , wherein the recombinant GH61 polypeptide comprises the amino acid sequence of SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 28, SEQ ID NO: 30, or SEQ ID NO: 90.

33 - 37 : (canceled)

38 . The method of claim 19 , wherein the recombinant CDH-heme domain polypeptide comprises the amino acid sequence of SEQ ID NO: 32 or SEQ ID NO: 46.

39 . The method of claim 19 , wherein the CDH-heme domain comprises the amino acid sequence selected from the group consisting of SEQ ID NO: 70, SEQ ID NO: 76, SEQ ID NO: 80, and SEQ ID NO: 86, and wherein the CBM comprises the amino acid sequence of SEQ ID NO: 74 or SEQ ID NO: 84.

40 . The method of claim 19 , wherein the method further comprises having a concentration of between 0.1-500 μM copper in the reaction mixture.

41 . The method of claim 40 , wherein the concentration of copper in the reaction mixture is 1-50 μM.

42 . The method of claim 19 , wherein the recombinant GH61 polypeptide comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 28, SEQ ID NO: 30, or SEQ ID NO: 90.

43 . The method of claim 19 , wherein the recombinant CDH-heme domain polypeptide comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 32 or SEQ ID NO: 46.

44 . The method of claim 19 , wherein the CDH-heme domain comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence selected from the group consisting of SEQ ID NO: 70, SEQ ID NO: 76, SEQ ID NO: 80, and SEQ ID NO: 86, and wherein the CBM comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 74 or SEQ ID NO: 84.

45 . The method of claim 19 , wherein the recombinant GH61 polypeptide comprises the motif H-X (4-8) -Q-X-Y.

46 . The method of claim 19 , wherein the recombinant CDH-heme domain polypeptide comprises a first domain and a second domain, wherein the first domain comprises a CDH-heme domain and the second domain comprises a CBM, and wherein the polypeptide does not contain a dehydrogenase domain.

47 . The method of claim 19 , wherein the recombinant CDH-heme domain polypeptide comprises a first domain, a second domain, and a third domain, wherein the first domain comprises a CDH-heme domain, the second domain comprises a CBM, and the third domain comprises a dehydrogenase domain.

Assignees

• Univ California

Inventors

• Marletta Michael A
• Doudna Cate James H
• Beeson Iv William T
• Phillips Christopher M

Classifications

• C12N9/0006 Primary

  acting on CH-OH groups as donors (1.1) · CPC title

• C12P19/14 Primary

  produced by the action of a carbohydrase {(EC 3.2.x)}, e.g. by alpha-amylase {, e.g. by cellulase, hemicellulase} · CPC title

• C12P19/02

  Monosaccharides (2-ketogulonic acid C12P7/60) · CPC title

• D21C5/005

  Treatment of cellulose-containing material with microorganisms or enzymes · CPC title

• C07K2319/00

  Fusion polypeptide · CPC title