# US 2013/0280786 A1 · Enzymes and uses thereof

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- Number: US 2013/0280786 A1
- Document ID: US20130280786A1
- Status: Published application
- Kind: A1
- Application number: 13/884,087
- Filed: 2011-11-08
- Published: 2013-10-24
- Earliest claimed: 2010-11-08
- Assignees: Deinove
- Inventors: Jean-Paul Leonetti; Jean-Michel Claverie; Pascale Joseph; Lucie Roux
- Web page: https://patentgrep.com/patent/US20130280786A1

## CPC

- C12N 15/74: Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora
- C12N 9/0006: {acting on CH-OH groups as donors (1.1)}
- C12N 9/0008: {acting on the aldehyde or oxo group of donors (1.2)}
- C12N 9/16: acting on ester bonds (3.1)
- C12N 9/18: Carboxylic ester hydrolases {(3.1.1)}
- C12N 9/2402: {hydrolysing O- and S- glycosyl compounds (3.2.1)}
- C12N 9/2411: {Amylases}
- C12N 9/2417: {from microbiological source}
- C12N 9/2428: {Glucan 1,4-alpha-glucosidase (3.2.1.3), i.e. glucoamylase}
- C12N 9/2437: {Cellulases (3.2.1.4; 3.2.1.74; 3.2.1.91; 3.2.1.150)}
- C12N 9/244: {Endo-1,3(4)-beta-glucanase (3.2.1.6)}
- C12N 9/248: {Xylanases}
- C12N 9/2497: {hydrolysing N- glycosyl compounds (3.2.2)}
- C12P 1/04: by using bacteria
- C12P 7/065: {with microorganisms other than yeasts}
- 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}
- C12P 39/00: Processes involving microorganisms of different genera in the same process, simultaneously
- C12Y 301/01072: Acetylxylan esterase (3.1.1.72)
- C12Y 101/01001: Alcohol dehydrogenase (1.1.1.1)
- C12Y 102/0101: Acetaldehyde dehydrogenase (acetylating) (1.2.1.10)
- C12Y 302/01003: Glucan 1,4-alpha-glucosidase (3.2.1.3), i.e. glucoamylase
- C12Y 302/01004: Cellulase (3.2.1.4), i.e. endo-1,4-beta-glucanase
- C12Y 302/01008: Endo-1,4-beta-xylanase (3.2.1.8)
- Y02E 50/10: Biofuels, e.g. bio-diesel
- 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
- Y02E: REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION

## Abstract

The present invention relates to novel enzymes and the uses thereof. The invention also relates to methods of producing such enzymes, coding nucleic acid molecules, recombinant cells and methods of transforming biomass from such materials. The invention is particularly suited to degrade biomass and/or to improve biomass degradation, and to produce bioenergy products or recombinant proteins. This invention also relates to various applications of the enzymes in the field of paper industry, textile industry as well as in the chemical and medical fields.

## Claims

### Claim at position 1

1-18. (canceled)

### Claim 19

An isolated enzyme or polypeptide selected from:
a) an enzyme, wherein said enzyme is derived from a Deinococcus or a related bacterium and is involved in energetic metabolism; or
b) a polypeptide comprising an amino acid sequence selected from SEQ ID NOs: 1-12, 27-41, 58, 60, 62, 64, 66, 68, 70 or 72 or a fragment thereof comprising at least 15 contiguous residues thereof.

### Claim 20 (depends on 19)

The isolated enzyme of claim 19, which is active at a temperature of 30° C. or more.

### Claim 21 (depends on 19)

The isolated enzyme of claim 19, wherein said enzyme catalyses biomass modification.

### Claim 22 (depends on 21)

The isolated enzyme of claim 21, wherein said enzyme is selected from amylases, glucosidases, cellulases, xylanases, pectinases, esterases, acetyl xylan esterases, or glucuronidases.

### Claim 23 (depends on 22)

The isolated enzyme of claim 22, which comprises all or an active part of an amino acid sequence selected from SEQ ID NOs: 1-12, 58, 60, 62, 64, 66, 68, 70 or 72.

### Claim 24 (depends on 21)

The isolated enzyme of claim 21, wherein said enzyme is involved in biofuel production by fermentation.

### Claim 25 (depends on 24)

The isolated enzyme of claim 24, wherein said enzyme is selected from acetaldehyde dehydrogenases, alcohol dehydrogenases or pyruvate dehydrogenases.

### Claim 26 (depends on 24)

The isolated enzyme of claim 24, which comprises all or an active part of an amino acid sequence selected from SEQ ID NOs: 27-41.

### Claim 27

A isolated polypeptide comprising an amino acid sequence selected from SEQ ID NOs: 1-12, 27-41, 58, 60, 62, 64, 66, 68, 70 or 72 or a fragment thereof comprising at least 15 contiguous residues thereof.

### Claim 28 (depends on 19)

An isolated nucleic acid coding an enzyme or polypeptide of claim 19.

### Claim 29 (depends on 28)

A vector comprising a nucleic acid of claim 28.

### Claim 30 (depends on 28)

A recombinant cell containing a nucleic acid of claim 28.

### Claim 31 (depends on 30)

The recombinant cell of claim 30, wherein said recombinant cell comprises a vector comprising said nucleic acid.

### Claim 32 (depends on 30)

The recombinant cell of claim 30, which is a Deinococcus or a related bacterium.

### Claim 33 (depends on 19)

A method for modifying biomass, comprising exposing biomass to an enzyme of claim 19.

### Claim 34 (depends on 33)

The method of claim 33, wherein said biomass is exposed to a cell expressing said enzyme.

### Claim 35 (depends on 19)

A method for producing metabolites or bioenergy products, comprising exposing a carbon source to an enzyme of claim 19.

### Claim 36 (depends on 35)

The method of claim 35, wherein said carbon source is exposed to a cell expressing said enzyme.

### Claim 37 (depends on 19)

A co-culture of at least two distinct microorganisms, wherein at least one of said microorganisms is a Deinococcus bacterium and at least one of said microorganisms is a prokaryotic or eukaryotic cell, wherein said at least two microorganisms are symbiotic to each other, and wherein said at least one Deinococcus bacterium exhibits an enzymatic activity according to claim 19.

### Claim 38 (depends on 37)

The co-culture of claim 37, wherein at least one of said microorganisms is eukaryotic and is a yeast cell.