# US 2017/0280742 A1 · Method of fermenting cocoa beans

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- Number: US 2017/0280742 A1
- Document ID: US20170280742A1
- Status: Published application
- Kind: A1
- Application number: 15/507,505
- Filed: 2015-08-27
- Published: 2017-10-05
- Earliest claimed: 2014-08-29
- Assignees: Dupont Nutrition Biosciences Aps
- Applicants: Dupont Nutrition Biosciences Aps
- Inventors: PASCAL FOURCASSIE; PATRICK BOYAVAL; CLAIRE BONARD
- Web page: https://patentgrep.com/patent/US20170280742A1

## CPC

- A23G 1/02: Preliminary treatment, e.g. fermentation of cocoa
- A23G 1/003: {with introduction or production of gas, or under vacuum; Whipping; Manufacture of cellular mass}
- A23G 1/56: Liquid products; Solid products in the form of powders, flakes or granules for making liquid products, e.g. for making chocolate milk {, drinks and the products for their preparation, pastes for spreading or milk crumb (A23G1/305 takes precedence)}
- C12N 1/02: Separating microorganisms from their culture media
- C12N 1/16: Yeasts; Culture media therefor
- C12N 1/20: Bacteria; Culture media therefor
- A23G 1/00: Cocoa; Cocoa products, e.g. chocolate; Substitutes therefor (preservation thereof A23B80/00)
- A23G 1/0006: {Processes specially adapted for manufacture or treatment of cocoa or cocoa products (A23G1/002 - A23G1/005 take precedence)}
- C12N 1/00: Microorganisms; Compositions thereof (medicinal preparations containing material from protozoa, bacteria or viruses A61K35/66, from algae A61K36/02, from fungi A61K36/06; preparing medicinal bacterial antigen or antibody compositions, e.g. bacterial vaccines, A61K39/00); Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- Y02E 50/10: Biofuels, e.g. bio-diesel
- A23G: COCOA; COCOA PRODUCTS, e.g. CHOCOLATE; SUBSTITUTES FOR COCOA OR COCOA PRODUCTS; CONFECTIONERY; CHEWING GUM; ICE-CREAM; PREPARATION THEREOF
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- Y02E: REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION

## Abstract

Use of a yeast capable of producing ethanol and acetic acid by fermentation of a carbohydrate, in a method for the fermentation of cocoa beans, is disclosed.

## Claims

### Claim 1

A method for the fermentation of cocoa beans comprising the steps of:
a) adding at least one yeast to a plant material comprising:
(i) beans derived from fruit pods of the species Theobroma cacao; and
(ii) pulp; and
b) fermenting the plant material to obtain fermented cocoa beans, the fermentation environment containing at least one other microorganism selected from the group consisting of yeast, lactic acid bacteria and acetic acid bacteria;
wherein the added yeast is able to produce ethanol and acetic acid by fermentation of a carbohydrate.

### Claim 2 (depends on 1)

A method according to claim 1, wherein said added yeast is able:
to produce acetic acid during assay I in a concentration of at least 0.7 g/L, and/or
to produce acetic acid during assay I in a ratio of the concentration of acetic acid produced by said yeast over the final OD of at least 50 mg/L/OD.

### Claim 3 (depends on 1)

The method according to claim 1, wherein no acetic acid bacterium is added during the fermentation step.

### Claim 4 (depends on 1)

The method according to claim 1, wherein more than 60% of the acetic acid produced during the fermentation of cocoa beans is carried out by said at least one added yeast.

### Claim 5 (depends on 1)

The method according to claim 1, wherein the ratio (log/log) total yeast/total acetic acid bacteria present in the fermentation environment is more than 0.8 during the 3 first days of fermentation.

### Claim 6 (depends on 1)

The method according to claim 1, wherein said at least one yeast is from a genus selected from the group consisting of Dekkera, Brettanomyces, Hanseniaspora, Kloeckera, Pichia, Candida, Saccharomyces, Kluyveromyces, Debaryomyces, Kazachstania, Wickerhamomyces, Lindnera and Zygotorulaspora, or any mixture thereof.

### Claim 7 (depends on 1)

The method according to claim 1, wherein said at least one yeast is from a species selected from the group consisting of Dekkera bruxellensis (anamorph Brettanomyces bruxellensis), Brettanomyces custersianus, Brettanomyces nanus, Brettanomyces naardenensis, Hanseniaspora uvarum (anamorph Kloeckera apiculata), Hanseniaspora guillermondii (anamorph Kloeckera apis), Hanseniaspora osmophila (anamorph Kloeckera cortisis), Pichia fermentans (anamorph Candida lambica), Pichia membranifaciens (anamorph Candida valida), Wickerhamomyces anomalus, Pichia kluyveri, Kluyveromyces lactis, Debaryomyces hansenii (anamorph Candida famata), Lindnera jadinii (anamorph Candida utilis), Kazachstania unispora, Zygotorulaspora florentina, and Saccharomyces bayanus or a mixture of any thereof.

### Claim 8 (depends on 1)

The method according to claim 1, wherein said at least one yeast has a pectinolytic activity.

### Claim 9 (depends on 1)

The method according to claim 1, wherein:
at least one other yeast is added in step (a), and
said other yeast has a pectinolytic activity.

### Claim 10 (depends on 1)

The method according to claim 1, wherein at least one enzyme having a pectinolytic activity is added in step (a).

### Claim 11 (depends on 1)

A method of producing a cocoa-based product, wherein the method comprises:
(a) providing fermented cocoa beans obtained or obtainable by a method according to claim 1, and
(b) carrying out one or more further processing steps on said fermented cocoa beans to produce the cocoa-based product.

### Claim 12 (depends on 11)

The method according to claim 11 wherein the one more more further processing steps of (b) comprise(s) drying said fermented cocoa beans.

### Claim 13 (depends on 11)

The method according to claim 11, wherein the method comprises a further step of thermally treating the dried beans.

### Claim 14 (depends on 13)

The method according to claim 13, wherein the method comprises further steps of:
removing shells from the roasted beans to produce cocoa nibs, and
grinding the nibs to produce a cocoa liquor.

### Claim 15

(canceled)

### Claim 16

A mixture of yeasts, wherein the mixture comprises at least 2 yeasts of different species selected independently from the group consisting of Saccharomyces bayanus, Saccharomyces cerevisiae, Kluyveromyces marxianus (Candida kefyr), Dekkera bruxellensis, Hanseniaspora guillermondii, Pichia kluyveri and Pichia anomala.

### Claim 17 (depends on 16)

A mixture of yeasts according to claim 16, wherein:
one yeast is selected from the group consisting of Saccharomyces bayanus and Dekkera bruxellensis; and
one yeast is selected from the group consisting of Saccharomyces bayanus, Dekkera bruxellensis, Kluyveromyces marxianus (Candida kefyr), Saccharomyces cerevisiae, Hanseniaspora guillermondii, Pichia kluyveri and Pichia anomala.

### Claim 18 (depends on 1)

A method according to claim 1, wherein said added yeast is able:
to produce ethanol during assay I in a concentration of at least 30 g/L, and/or
to produce ethanol during assay I in a ratio of the concentration of ethanol produced by said yeast over the final OD of at least 3 g/L/OD.

### Claim 19 (depends on 1)

The method according to claim 1, wherein the ratio (log/log) total yeast/total acetic acid bacteria present in the fermentation environment is more than 0.8 during the whole fermentation step.

### Claim 20 (depends on 1)

The method according to claim 1, wherein said at least one yeast is a species selected from the group consisting of Dekkera bruxellensis and Saccharomyces bayanus.

### Claim 21 (depends on 1)

The method according to claim 1, wherein said at least one yeast is selected from the group consisting of a mixture of yeasts comprising Dekkera bruxellensis and a mixture of yeasts comprising Saccharomyces bayanus.

### Claim 22 (depends on 7)

The method according to claim 7, wherein any yeast species from Pichia kluyveri is other than the strains Pichia kluyveri PK-KR1 (V06/022711) and Pichia kluyveri PKKR2 (V06/022712).