# US 2019/0169655 A1 · Methods of producing lactic acid or a salt or an ester thereof by using a versatile extremely thermophilic bacteria

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- Number: US 2019/0169655 A1
- Document ID: US20190169655A1
- Status: Published application
- Kind: A1
- Application number: 16/271,674
- Filed: 2019-02-08
- Published: 2019-06-06
- Earliest claimed: 2011-10-07
- Assignees: DIREVO Industrial Biotechnology GmbH
- Applicants: DIREVO Industrial Biotechnology GmbH
- Inventors: Vitaly Svetlichnyi; Simon Curvers
- Web page: https://patentgrep.com/patent/US20190169655A1

## CPC

- C12P 7/065: {with microorganisms other than yeasts}
- C12N 1/20: Bacteria; Culture media therefor
- C12N 1/205: Bacterial isolates
- C12P 7/10: substrate containing cellulosic material
- C12P 7/14: Multiple stages of fermentation; Multiple types of microorganisms or re-use of microorganisms
- C12P 7/54: Acetic acid (vinegar C12J)
- C12P 7/56: Lactic acid
- C12P 2201/00: Pretreatment of cellulosic or lignocellulosic material for subsequent enzymatic treatment or hydrolysis
- C12P 2203/00: Fermentation products obtained from optionally pretreated or hydrolyzed cellulosic or lignocellulosic material as the carbon source
- C12R 2001/01: Bacteria or Actinomycetales {; using bacteria or Actinomycetales}
- 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)
- C12R: INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- Y02E: REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION

## Abstract

A method of producing lactic acid or a salt or an ester thereof, which includes incubating lignocellulosic hydrolysates in the presence of cells of an isolated strain of Thermoanaerobactor at a temperature above 70 degrees Celsius, wherein the Thermoanaerobactor comprises a 16S rDNA sequence at least 99% identical to the nucleic acid sequence selected from the group consisting of SEQ ID NO. 1, SEQ ID NO. 2, SEQ ID NO. 3, SEQ ID NO. 4, SEQ ID NO. 5, SEQ ID NO. 6, SEQ ID NO. 7, and SEQ ID NO. 8

## Claims

### Claim 1

A method of producing lactic acid or a salt or an ester thereof, the method comprising incubating lignocellulosic hydrolysates in the presence of cells of an isolated strain of Thermoanaerobactor at a temperature above 70 degrees Celsius, wherein the Thermoanaerobactor comprises a 16S rDNA sequence at least 99% identical to the nucleic acid sequence selected from the group consisting of SEQ ID NO. 1, SEQ ID NO. 2, SEQ ID NO. 3, SEQ ID NO. 4, SEQ ID NO. 5, SEQ ID NO. 6, SEQ ID NO. 7, and SEQ ID NO. 8.

### Claim 2 (depends on 1)

The method of claim 1, wherein the lignocellulosic hydrolysates are obtained by providing a lignocellulosic biomass and subjecting the biomass to a treatment that separates the biomass into cellulose, hemicellulose and lignin.

### Claim 3 (depends on 2)

The method of claim 2, wherein the biomass is a grass, optionally selected from the group consisting of switch grass, cord grass, rye grass, reed canary grass, mixed prairie grass, and miscanthus.

### Claim 4 (depends on 2)

The method of claim 2, wherein the biomass is a straw, optionally selected from the group consisting of sugarcane straw, rice straw, barley straw, cereal straw, wheat straw, canola straw, and oat straw.

### Claim 5 (depends on 2)

The method of claim 2, wherein the biomass is selected from the group consisting of agricultural waste, corn cobs, oat hulls, corn fiber, stover, soybean stover, corn stover, cotton stalks, forestry wastes, recycled wood pulp fiber, paper sludge, sawdust, sugar-methoding residues, sugarcane bagasse, and rice hulls.

### Claim 6 (depends on 2)

The method of claim 2, wherein the biomass is a hardwood or a softwood.

### Claim 7 (depends on 2)

The method of claim 2, wherein the treatment is selected from the group consisting of a mechanical treatment, a thermochemical treatment, and a biochemical treatment.

### Claim 8 (depends on 7)

The method of claim 7, wherein the treatment is mechanical comminution, the method further comprising performing a subsequent treatment with sulfurous acid or its anhydride under increased heat and pressure, optionally with a sudden release of pressure.

### Claim 9 (depends on 7)

The method of claim 7, wherein the treatment is mechanical comminution, the method further comprising performing a subsequent treatment with a chemical under increased heat and pressure, optionally with a sudden release of pressure, wherein the chemical is selected from the group consisting of sodium hydroxide, ammonium hydroxide, calcium hydroxide, and potassium hydroxide.

### Claim 10 (depends on 7)

The method of claim 7, wherein the treatment is the mechanical comminution, the method further comprising steaming the biomass, separating partially and fully solubilized components from insoluble components, and providing the insoluble components as the lignocellulosic hydrolysates.

### Claim 11 (depends on 2)

The method of claim 2, wherein the treatment is a steam treatment.

### Claim 12 (depends on 11)

The method of claim 11, wherein the steam treatment comprises sulfurous acid, the treatment further comprising an enzymatic treatment.

### Claim 13 (depends on 12)

The method of claim 12, wherein the enzymatic treatment comprises treatment with a cellulose, a glucosidase, and/or a hemicellulase.

### Claim 14 (depends on 1)

The method of claim 1, wherein the strain is selected from the group consisting of DIB004G, DIB097G, DIB101G, DIB101X, DIB087G, DIB103X, DIB104X and DIB107X, wherein: DIB004G is DSMZ Accession No. 25179, DIB097G is DSMZ Accession No. 25308, DIB101G is DSMZ Accession No. 25180, DIB101X is DSMZ Accession No. 25181, DIB087G is DSMZ Accession No. 25777, DIB103X is DSMZ Accession No. 25776, DIB104X is DSMZ Accession No. 25778, and DIB107X is DSMZ Accession No. 25779.

### Claim 15 (depends on 14)

The method of claim 14 wherein the strain is DIB004G (DSMZ Accession No. 25179).

### Claim 16 (depends on 1)

The method of claim 1, wherein the 16S rDNA sequence comprises the nucleic acid sequence of SEQ ID NO. 1.