# US 10,479,983 B2 · Variant enzymes

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 10,479,983 B2
- Document ID: US10479983B2
- Status: Granted
- Kind: B2
- Application number: 14/909,041
- Filed: 2014-07-24
- Published: 2016-06-30
- Granted: 2019-11-19
- Earliest claimed: 2013-07-29
- Assignees: Danisco US Inc.
- Applicants: Danisco US Inc
- Inventors: Nicholai Douglas; Maria Foukaraki; Ronaldus Wilhelmus Hommes; Thijs Kaper; Bradley R. Kelemen; Slavko Kralj; Suzanne E. Lantz; Jonathan K. Lassila; Igor Nikolaev; Daniel Esteban Torres-Pazmino; Wilhelmus Van Der Kley; Johannes Franciscus Thomas Van Lieshout; Sander Van Stigt Thans
- Web page: https://patentgrep.com/patent/US10479983B2

## CPC

- C12N 9/2437: {Cellulases (3.2.1.4; 3.2.1.74; 3.2.1.91; 3.2.1.150)}
- C12N 9/0083: {Miscellaneous (1.14.99)}
- C12N 9/2402: {hydrolysing O- and S- glycosyl compounds (3.2.1)}
- C12N 9/2434: {acting on beta-1,4-glucosidic bonds}
- C12P 19/02: Monosaccharides (2-ketogulonic acid C12P7/60)
- C12P 19/14: produced by the action of a carbohydrase {(EC 3.2.x)}, e.g. by alpha-amylase {, e.g. by cellulase, hemicellulase}
- C12Y 302/01004: Cellulase (3.2.1.4), i.e. endo-1,4-beta-glucanase
- 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

## Abstract

Disclosed are glycosyl hydrolase enzyme variants, particularly variants of certain oxidoreductases of glycosyl hydrolase family 61. Nucleic acids encoding the glycosyl hydrolyase variants, compositions including the glycosyl hydrolase variants, methods of producing the variants, and methods of using the variants are also described.

## Claims

### Claim 1

A variant glycoside hydrolase family 61 (GH61) polypeptide, wherein said variant has cellulase enhancing activity, has at least 95% sequence identity to the amino acid sequence of SEQ ID NO:3 and the amino acid substitution I114L, and has at least one improved property relative to a parent GH61 polypeptide, wherein said improved property is selected from the group consisting of (a) expression based on protein level and RNA level or activity of the variant GH61 polypeptide, (b) thermostability and/or melting temperature (Tm), (c) performance in a Whole Hydrolysate Dilute Acid Pretreated Corn Stover (whPCS) hydrolysis assay.

### Claim 2 (depends on 1)

The variant of claim 1, wherein said variant is a combinatorial variant.

### Claim 3 (depends on 1)

The variant of claim 1, wherein said parent GH61 polypeptide is a fungal glycosyl hydrolase 61a (GH61A).

### Claim 4 (depends on 3)

The variant of claim 3, wherein said fungal GH61A is from Hypocrea jecorina, Hypocrea rufa, Hypocrea orientalis, Hypocrea atroviridis, Hypocrea virens, Emericella nidulans, Aspergillus terreus, Aspergillus oryzae, Aspergillus niger, Aspergillus kawachii, Aspergillus flavus, Aspergillus clavatus, Gaeumannomyces graminis, Trichoderma saturnisporum, Neurospora tetrasperma, Neurospora crassa, Neosartorya fumigata, Neosartorya fumigate, Neosartorya fischeri, Thielavia terrestris, Talaromyces sp., Sporotricum sp, and Thielavia heterothallica.