Fungal proteases

Patent metadata

Field	Value
Publication number	US-2016333329-A1
Application number	US-201615226522-A
Country	US
Kind code	A1
Filing date	Aug 2, 2016
Priority date	Sep 30, 2011
Publication date	Nov 17, 2016
Grant date	—

Abstract

The present invention provides fungal proteases and improved fungal strains that are deficient in protease production.

First claim

What is claimed is:

1 . A method for producing a genetically modified Myceliophthora deficient in at least one protease native to said Myceliophthora , comprising providing a Myceliophthora having protease activity, wherein said protease activity comprises lysing Myceliophthora cellobiohydrolase enzymes and said protease comprises an amino acid sequence having at least 98% identity with the polypeptide sequence set forth in SEQ ID NO:3, 6, 9, or 12; and mutating said Myceliophthora under conditions such that said protease is mutated to produce a protease-deficient Myceliophthora.

2 . The method of claim 1 , wherein said Myceliophthora is Myceliophthora thermophila.

3 . The method of claim 1 , wherein said Myceliophthora further produces at least one cellulase selected from endoglucanases, cellobiohydrolases, cellobiose dehydrogenases, endoxylanases, beta-xylosidases, arabinofuranosidases, alpha-glucuronidases, acetylxylan esterases, feruloyl esterases, and/or alpha-glucuronyl esterases.

4 . The method of claim 3 , wherein said cellulase is a recombinant cellulase selected from EG1b, EG2, EG3, EG4, EG5, EG6, CBH1a, CBH1b, CBH2a, CBH2b, GH61a, GH61f, and/or GH61p.

5 . The method of claim 1 , wherein at least one polynucleotide sequence selected from SEQ ID NOS:1, 2, 4, 5, 7, 8, 10, or 11 have been deleted from the genome of said Myceliophthora , such that the Myceliophthora produces a reduced level of at least one protease, as compared to a wild-type Myceliophthora.

6 . The method of claim 1 , wherein at least one polynucleotide sequence selected from SEQ ID NOS:1, 2, 4, 5, 7, 8, 10, or 11 have been mutated in the genome of said Myceliophthora , such that the Myceliophthora produces a reduced level of at least one protease, as compared to a wild-type Myceliophthora.

7 . The method of claim 5 , wherein said Myceliophthora is Myceliophthora thermophila.

8 . The method of claim 6 , wherein said Myceliophthora is Myceliophthora thermophila.

Assignees

• Codexis Inc

Inventors

• Szabo Lorand
• Molnar Zsolt
• Nemeth Attila Laszlo

Classifications

• C12N9/58 Primary

  derived from fungi · CPC title

• C12N9/48

  acting on peptide bonds (3.4) · CPC title

• C12N1/14

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

• C12P21/02

  having a known sequence of two or more amino acids, e.g. glutathione · CPC title

• C12N9/2482

  Endo-1,4-beta-xylanase (3.2.1.8) · CPC title