# US 9,434,919 B2 · Fungal proteases

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- Number: US 9,434,919 B2
- Document ID: US9434919B2
- Status: Granted
- Kind: B2
- Application number: 14/950,712
- Filed: 2015-11-24
- Published: 2016-03-17
- Granted: 2016-09-06
- Earliest claimed: 2011-09-30
- Assignees: Codexis, Inc.
- Applicants: Codexis, Inc.
- Inventors: Lorand Szabo; Zsolt Molnar; Attila Laszlo Nemeth
- Web page: https://patentgrep.com/patent/US9434919B2

## CPC

- C12N 15/1137: {against enzymes (viral enzymes C12N15/1131; receptors C12N15/1138)}
- C12N 1/14: Fungi (culture of mushrooms A01G18/00; as new plants A01H15/00); Culture media therefor
- C12N 1/145: Fungi isolates
- C12N 9/20: Triglyceride splitting, e.g. by means of lipase
- C12N 9/24: acting on glycosyl compounds (3.2)
- C12N 9/2482: {Endo-1,4-beta-xylanase (3.2.1.8)}
- C12N 9/48: acting on peptide bonds (3.4)
- C12N 9/58: derived from fungi
- C12N 15/80: for fungi
- C12P 19/02: Monosaccharides (2-ketogulonic acid C12P7/60)
- C12P 21/00: Preparation of peptides or proteins (single cell protein C12N1/00)
- C12P 21/02: having a known sequence of two or more amino acids, e.g. glutathione
- C12Y 301/01008: Cholinesterase (3.1.1.8), i.e. butyrylcholine-esterase
- C12Y 304/00: Hydrolases acting on peptide bonds, i.e. peptidases (3.4)
- C12P 19/14: produced by the action of a carbohydrase {(EC 3.2.x)}, e.g. by alpha-amylase {, e.g. by cellulase, hemicellulase}
- C12R 2001/645: Fungi {; Processes using fungi}
- Y02P 20/52: using catalysts, e.g. selective catalysts
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12P: FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {(brewing of beer C12C; producing vinegar C12J; producing specific peptides or proteins C07K; producing enzymes C12N9/00; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; measuring or testing processes involving enzymes or microorganisms C12Q; measuring or testing processes involving nucleic acid amplification reactions C12Q1/6844; fermentation processes to form a food composition, A21 or A23; compounds in general, see the relevant compound class, e.g. C01, C07)}
- C12Y: ENZYMES
- C12R: INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- Y02P: CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS

## Abstract

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

## Claims

### Claim 1

An isolated, genetically modified Myceliophthora deficient in at least one protease native to said Myceliophthora, wherein said protease is capable of degrading Myceliophthora cellobiohydrolase enzymes, and comprises an amino acid sequence having at least 98% identity with the polypeptide sequence set forth in SEQ ID NO:12.

### Claim 2 (depends on 1)

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

### Claim 3 (depends on 1)

The Myceliophthora of claim 1, wherein said Myceliophthora produces at least one enzyme.

### Claim 4 (depends on 3)

The Myceliophthora of claim 3, wherein said at least one enzyme comprises at least one cellulase.

### Claim 5 (depends on 4)

The Myceliophthora of claim 4, wherein said cellulase is beta-glucosidase.

### Claim 6 (depends on 5)

The Myceliophthora of claim 5, wherein said cellulase is a recombinant beta-glucosidase.

### Claim 7 (depends on 4)

The Myceliophthora of claim 4, wherein said Myceliophthora further produces at least one non-cellulase enzyme.

### Claim 8 (depends on 7)

The Myceliophthora of claim 7, wherein said at least one non-cellulase enzyme comprises at least one lipase, amylase, glucoamylase, and/or protease.

### Claim 9 (depends on 1)

A composition comprising the Myceliophthora of claim 1.