Aspergillus fumigatus cellulolytic enzyme compositions and uses thereof

Patent metadata

Field	Value
Publication number	US-9970015-B2
Application number	US-201715489035-A
Country	US
Kind code	B2
Filing date	Apr 17, 2017
Priority date	Aug 24, 2011
Publication date	May 15, 2018
Grant date	May 15, 2018

Abstract

The present invention relates to recombinant Trichoderma host cells producing Aspergillus fumigatus cellulolytic enzyme compositions and methods of producing and using the compositions.

First claim

What is claimed is:

1. An enzyme composition comprising a recovered fermentation broth of a recombinant Trichoderma host cell, wherein the recombinant Trichoderma host cell comprises polynucleotides encoding: (i) an Aspergillus fumigatus cellobiohydrolase I; (ii) an Aspergillus fumigatus cellobiohydrolase II; (iii) an Aspergillus fumigatus beta-glucosidase; (iv) an Aspergillus fumigatus GH61 polypeptide having cellulolytic enhancing activity; (v) an Aspergillus fumigatus xylanase; and (vi) an Aspergillus fumigatus beta-xylosidase; or homologs thereof;
wherein the Aspergillus fumigatus cellobiohydrolase I or homolog thereof is selected from the group consisting of:
(i) a cellobiohydrolase I comprising the mature polypeptide of SEQ ID NO: 2;
(ii) a cellobiohydrolase I comprising an amino acid sequence having at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 2;
(iii) a cellobiohydrolase I encoded by a polynucleotide comprising a nucleotide sequence having at least 95% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 1; and
(iv) a cellobiohydrolase I encoded by a polynucleotide that hybridizes under very high stringency conditions with the mature polypeptide coding sequence of SEQ ID NO: 1 or the full-length complement thereof;
wherein the Aspergillus fumigatus cellobiohydrolase II or homolog thereof is selected from the group consisting of:
(i) a cellobiohydrolase II comprising the mature polypeptide of SEQ ID NO: 4;
(ii) a cellobiohydrolase II comprising an amino acid sequence having at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 4;
(iii) a cellobiohydrolase II encoded by a polynucleotide comprising a nucleotide sequence having at least 95% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 3; and
(iv) a cellobiohydrolase II encoded by a polynucleotide that hybridizes under very high stringency conditions with the mature polypeptide coding sequence of SEQ ID NO: 3 or the full-length complement thereof;
wherein the Aspergillus fumigatus beta-glucosidase or homolog thereof is selected from the group consisting of:
(i) a beta-glucosidase comprising the mature polypeptide of SEQ ID NO: 6;
(ii) a beta-glucosidase comprising an amino acid sequence having at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 6;
(iii) a beta-glucosidase encoded by a polynucleotide comprising a nucleotide sequence having at least 95% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 5; and
(iv) a beta-glucosidase encoded by a polynucleotide that hybridizes under very high stringency conditions with the mature polypeptide coding sequence of SEQ ID NO: 5 or the full-length complement thereof;
wherein the Aspergillus fumigatus GH61 polypeptide having cellulolytic enhancing activity or homolog thereof is selected from the group consisting of:
(i) a GH61 polypeptide having cellulolytic enhancing activity comprising the mature polypeptide of SEQ ID NO: 8;
(ii) a GH61 polypeptide having cellulolytic enhancing activity comprising an amino acid sequence having at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 8;
(iii) a GH61 polypeptide having cellulolytic enhancing activity encoded by a polynucleotide comprising a nucleotide sequence having at least 95% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 7; and
(iv) a GH61 polypeptide having cellulolytic enhancing activity encoded by a polynucleotide that hybridizes under very high stringency conditions with the mature polypeptide coding sequence of SEQ ID NO: 7 or the full-length complement thereof;
wherein the Aspergillus fumigatus xylanase or homolog thereof is selected from the group consisting of:
(i) an Aspergillus fumigatus xylanase comprising the mature polypeptide of SEQ ID NO: 14, SEQ ID NO: 16, or SEQ ID NO: 18;
(ii) a xylanase comprising an amino acid sequence having at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 14, SEQ ID NO: 16, or SEQ ID NO: 18;
(iii) a xylanase encoded by a polynucleotide comprising a nucleotide sequence having at least 95% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 13, SEQ ID NO: 15, or SEQ ID NO: 17; and
(iv) a xylanase encoded by a polynucleotide that hybridizes under very high stringency conditions with the mature polypeptide coding sequence of SEQ ID NO: 13, SEQ ID NO: 15, or SEQ ID NO: 17; or the full-length complement thereof;
wherein the Aspergillus fumigatus beta-xylosidase or homolog thereof is selected from the group consisting of:
(i) a beta-xylosidase comprising the mature polypeptide of SEQ ID NO: 20;
(ii) a beta-xylosidase comprising an amino acid sequence having at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 20;
(iii) a beta-xylosidase encoded by a polynucleotide comprising a nucleotide sequence having at least 95% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 19; and
(iv) a beta-xylosidase encoded by a polynucleotide that hybridizes under very high stringency conditions with the mature polypeptide coding sequence of SEQ ID NO: 19 or the full-length complement thereof; and

wherein very high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 micrograms/ml sheared and denatured salmon sperm DNA, and 50% formamide, and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 70° C.

2. The enzyme composition of claim 1 , wherein the recombinant Trichoderma host cell further comprises one or more polynucleotides encoding one or more enzymes selected from the group consisting of: (i) an Aspergillus fumigatus endoglucanase I; (ii) an Aspergillus fumigatus endoglucanase II; and (iii) an Aspergillus fumigatus swollenin;
wherein the Aspergillus fumigatus endoglucanase I or homolog thereof is selected from the group consisting of:
(i) an endoglucanase I comprising the mature polypeptide of SEQ ID NO: 10;
(ii) an endoglucanase I comprising an amino acid sequence having at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 10;
(iii) an endoglucanase I encoded by a polynucleotide comprising a nucleotide sequence having at least 95% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 9; and
(iv) an endoglucanase I encoded by a polynucleotide that hybridizes under very high stringency conditions with the mature polypeptide coding sequence of SEQ ID NO: 9 or the full-length complement thereof;
wherein the Aspergillus fumigatus endoglucanase II or homolog thereof is selected from the group consisting of:
(i) an Aspergillus fumigatus endoglucanase II comprising the mature polypeptide of SEQ ID NO: 12;
(ii) an endoglucanase II comprising an amino acid sequence having at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 12;
(iii) an endoglucanase II encoded by a polynucleotide comprising a nucleotide sequence having at least 95% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 11; and
(iv) an endoglucanase II encoded by a polynucleotide that hybridizes under very high stringency conditions with the mature polypeptide coding sequence of SEQ ID NO: 11 or the full-length complement thereof;
(i) wherein the Aspergillus fumigatus swollenin or homolog thereof is selected from the group consisting of:
(i) a swollenin comprising the mature polypeptide of SEQ ID NO: 22;
(ii) a swollenin comprising an amino acid sequence having at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 22;
(iii) a swollenin encoded by a polynucleotide comprising a nucleotide sequence having at least 95% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 21; and
(iv) a swollenin encoded by a polynucle

Assignees

• Novozymes Inc

Inventors

• Shasky Jeffrey
• Mcbrayer Brett

Classifications

• C12N9/248

  Xylanases · CPC title

• C12N1/14

  Fungi (culture of mushrooms A01G18/00; as new plants A01H15/00); Culture media therefor · CPC title

• C12N9/2405

  Glucanases · CPC title

• C12N9/2437

  Cellulases (3.2.1.4; 3.2.1.74; 3.2.1.91; 3.2.1.150) · CPC title

• C12N9/2485

  Xylan endo-1,3-beta-xylosidase (3.2.1.32), i.e. endo-1,3-beta-xylanase · CPC title