# US 2019/0316162 A1 · Processes for producing fermentation products

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- Number: US 2019/0316162 A1
- Document ID: US20190316162A1
- Status: Published application
- Kind: A1
- Application number: 16/434,550
- Filed: 2019-06-07
- Published: 2019-10-17
- Earliest claimed: 2013-09-11
- Assignees: Novozymes A/S
- Applicants: Novozymes A/S
- Inventors: Zhengfang Kang; Christie Strahler; Suzanne Clark; Kristian Bertel Rømer M. Krogh; Tang Lan
- Web page: https://patentgrep.com/patent/US20190316162A1

## CPC

- C12P 19/14: produced by the action of a carbohydrase {(EC 3.2.x)}, e.g. by alpha-amylase {, e.g. by cellulase, hemicellulase}
- 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 9/52: derived from bacteria {or Archaea}
- C12P 7/06: Ethanol, i.e. non-beverage
- C12P 7/14: Multiple stages of fermentation; Multiple types of microorganisms or re-use of microorganisms
- C12P 19/02: Monosaccharides (2-ketogulonic acid C12P7/60)
- C12P 2203/00: Fermentation products obtained from optionally pretreated or hydrolyzed cellulosic or lignocellulosic material as the carbon source
- C12Y 302/01001: Alpha-amylase (3.2.1.1)
- C12Y 302/01003: Glucan 1,4-alpha-glucosidase (3.2.1.3), i.e. glucoamylase
- C12Y 302/01006: Endo-1,3(4)-beta-glucanase (3.2.1.6)
- C12Y 304/24: Metalloendopeptidases (3.4.24)
- Y02E 50/10: Biofuels, e.g. bio-diesel
- 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)}
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12Y: ENZYMES
- Y02E: REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION

## Abstract

The present invention relates to processes for producing fermentation products from starch-containing material, wherein an alpha-amylase and a thermostable endoglucanase is present and/or added during liquefaction. The invention also relates to compositions suitable for use in processes of the invention.

## Claims

### Claim 1

A process for producing fermentation products from starch-containing material comprising the steps of:
i) liquefying the starch-containing material at a temperature from 75-95° at a pH from 4-6 using:
an alpha-amylase;
an endoglucanase having a Melting Point (DSC) above 80° C.;
ii) saccharifying using a carbohydrate-source generating enzyme; and
iii) fermenting using a fermenting organism.

### Claim 2 (depends on 1)

The process of claim 1, wherein the alpha-amylase has at least 90% sequence identity to the polypeptide of SEQ ID NO: 1.

### Claim 3 (depends on 2)

The process of claim 2, wherein the alpha-amylase has at least 95% sequence identity to the polypeptide of SEQ ID NO: 1.

### Claim 4 (depends on 1)

The process of claim 1, wherein the alpha-amylase has a T½ (min) at pH 4.5, 85° C., 0.12 mM CaCl2) of at least 10.

### Claim 5 (depends on 1)

The process of claim 1, further wherein a protease is present or added in liquefaction, wherein the protease has a thermostability value of more than 25% determined as Relative Activity at 80° C./70° C.

### Claim 6 (depends on 6)

The process of claim 6, wherein the protease has at least 90%, but less than 100%, sequence identity to the mature polypeptide of SEQ ID NO: 2.

### Claim 7 (depends on 6)

The process of claim 6, wherein the protease has at least 90% sequence identity to the polypeptide of SEQ ID NO: 13.

### Claim 8 (depends on 6)

The process of claim 6, wherein the protease has at least 95% sequence identity to the polypeptide of SEQ ID NO: 13.

### Claim 9 (depends on 1)

The process of claim 1, further wherein a carbohydrate-source generating enzyme is present or added during liquefaction step i).

### Claim 10 (depends on 9)

The process of claim 9, wherein the carbohydrate-source generating enzyme present or added during liquefaction step i) is a glucoamylase having a heat stability at 85° C., pH 5.3, of at least 20%.

### Claim 11 (depends on 9)

The process of claim 9, wherein the carbohydrate-source generating enzyme present or added during liquefaction step i) is a glucoamylase having at least 90% sequence identity to the polypeptide of SEQ ID NO: 9 or at least 90% sequence identity to the mature polypeptide of SEQ ID NO: 14.

### Claim 12 (depends on 9)

The process of claim 9, wherein the carbohydrate-source generating enzyme present or added during liquefaction step i) is a glucoamylase having at least 95% sequence identity to the polypeptide of SEQ ID NO: 9 or SEQ ID NO: 14.

### Claim 13

A process of producing liquefied starch, comprising liquefying a starch-containing material with a composition comprising:
i) an alpha-amylase;
ii) an endoglucanase having a Melting Point (DSC) above 80° C.;
iii) a protease; and
iv) optionally a carbohydrate-source generating enzyme.

### Claim 14 (depends on 13)

The process of claim 13, wherein the alpha-amylase has at least 90% sequence identity to the polypeptide of SEQ ID NO: 1.

### Claim 15 (depends on 13)

The process of claim 13, wherein the alpha-amylase has a T½ (min) at pH 4.5, 85° C., 0.12 mM CaCl2) of at least 10.

### Claim 16 (depends on 13)

The process of claim 13, wherein the protease has a thermostability value of more than 25% determined as Relative Activity at 80° C./70° C.

### Claim 17 (depends on 13)

The process of claim 13, wherein the protease has at least 90%, but less than 100%, sequence identity to the mature polypeptide of SEQ ID NO: 2.

### Claim 18 (depends on 13)

The process of claim 13, wherein the protease has at least 90% sequence identity to the polypeptide of SEQ ID NO: 13.

### Claim 19 (depends on 13)

The process of claim 13, wherein the carbohydrate-source generating enzyme is present and is a glucoamylase having a heat stability at 85° C., pH 5.3, of at least 20%.

### Claim 20 (depends on 19)

The process of claim 19, wherein the glucoamylase has at least 90% sequence identity to the polypeptide of SEQ ID NO: 9 or at least 90% sequence identity to the mature polypeptide of SEQ ID NO: 14.