Hetero-transglycosylase and uses thereof

Patent metadata

Field	Value
Publication number	US-10647965-B2
Application number	US-201816118956-A
Country	US
Kind code	B2
Filing date	Aug 31, 2018
Priority date	Sep 24, 2013
Publication date	May 12, 2020
Grant date	May 12, 2020

Abstract

The present invention relates to a hetero-transglycosylase protein having cellulose:xyloglucan endotransglucosylase (CXE) activity in addition to mixed-linkage beta-glucan:xyloglucan endotransglucosylase (MXE) activity. The protein may comprise the amino acid sequence of any one of SEQ ID NOs: 2, 6 and 8 or a functional fragment thereof; or an amino acid sequence having at least 60% sequence identity to any one of SEQ ID NO: 2, 6 and 8, or to SEQ ID NO: 2 from amino acid 22 to 280, to SEQ ID NO: 6 from amino acid 26 to 283, or to SEQ ID NO: 8 from amino acid 29 to 287. The invention furthermore relates to an isolated nucleic acid encoding the protein described herein, a chimeric gene comprising, inter alia, the nucleic acid described herein, a vector comprising said chimeric gene, a host cell comprising said vector or said chimeric gene and an according transgenic plant. Further disclosed herein in are a method of producing a transgenic plant and a method of improving properties of cellulosic material.

First claim

The invention claimed is:

1. A method for producing a protein having cellulose:xyloglucan endotransglucosylase activity comprising
1. the amino acid sequence of any one of SEQ ID NOs: 2, 6 and 8; or
2. an amino acid sequence having at least 95% sequence identity to the sequence of any one of SEQ ID NOs: 2, 6 and 8; or
3. an amino acid sequence having at least 95% sequence identity to the sequence of SEQ ID NO: 2 from amino acid 22 to 280, or to the sequence of SEQ ID NO: 6 from amino acid 26 to 283, or to the sequence of SEQ ID NO: 8 from amino acid 29 to 287,

comprising culturing a host cell comprising a chimeric gene comprising the following operably linked elements:
(a) a promoter,
(b) the nucleic acid encoding said protein and, optionally,
(c) a transcription termination and polyadenylation region;

and isolating the protein produced.

2. The method according to claim 1 , wherein said nucleic acid encoding said protein comprises a nucleic acid comprising:
a. a nucleic acid sequence having the sequence of any one of SEQ ID NOs: 1, 5 and 7; or
b. a nucleic acid sequence having the sequence of SEQ ID NO: 1 from nucleotide 64 to 840, or to the sequence of SEQ ID NO: 5 from nucleotide 76 to 849, or to the sequence of SEQ ID NO: 7 from nucleotide 85 to 861.

3. The method according to claim 1 , wherein said promoter is a constitutive promoter, a seed-specific promoter, a stem-specific promoter or a fiber-specific promoter.

4. The method of claim 1 , wherein the protein having cellulose:xyloglucan endotransglucosylase activity comprises: an amino acid sequence having at least 98% sequence identity to the sequence of any one of SEQ ID NOs: 2, 6 and 8; an amino acid sequence having at least 98% sequence identity to the sequence of SEQ ID NO: 2 from amino acid 22 to 280; or having at least 98% sequence identity to the sequence of SEQ ID NO: 6 from amino acid 26 to 283.

Assignees

• Basf Se
• Univ Court Univ Of Edinburgh

Inventors

• Meulewaeter Frank
• Van Den Brande Ilse
• Fry Stephen C
• Mohler Kyle E
• Frankova Lenka
• Simmons Tom J
• Holland Claire
• Hudson Andrew

Classifications

• C12N15/8246

  Non-starch polysaccharides, e.g. cellulose, fructans, levans · CPC title

• C08B37/0057

  beta-D-Xylans, i.e. xylosaccharide, e.g. arabinoxylan, arabinofuronan, pentosans; (beta-1,3)(beta-1,4)-D-Xylans, e.g. rhodymenans; Hemicellulose; Derivatives thereof · CPC title

• C08B1/00

  Preparatory treatment of cellulose for making derivatives thereof {, e.g. pre-treatment, pre-soaking, activation} · CPC title

• D06M15/03

  Polysaccharides or derivatives thereof · CPC title

• C12Y204/01207

  Xyloglucan:xyloglucosyl transferase (2.4.1.207) · CPC title