# US 10,767,200 B2 · Method for producing L-amino acid

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- Number: US 10,767,200 B2
- Document ID: US10767200B2
- Status: Granted
- Kind: B2
- Application number: 16/111,395
- Filed: 2018-08-24
- Published: 2019-02-07
- Granted: 2020-09-08
- Earliest claimed: 2016-02-24
- Assignees: Ajinomoto Co., Inc.
- Applicants: Ajinomoto Co., Inc.
- Inventors: Kisho Suzuki; Masayuki Mori; Takehiro Hiura; Nobuaki Matsuoka; Takehiko Chikamori
- Web page: https://patentgrep.com/patent/US10767200B2

## CPC

- C12P 13/08: Lysine; Diaminopimelic acid; Threonine; Valine
- C12N 1/005: {after treatment of microbial biomass not covered by C12N1/02 - C12N1/08}
- C12N 1/20: Bacteria; Culture media therefor
- C12N 1/205: Bacterial isolates
- C12R 2001/15: Corynebacterium
- C12R 2001/19: Escherichia coli
- 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

## Abstract

A method for producing a basic acid such as L-lysine is provided. A basic amino acid or a fermentation product containing the same is produced by a method including the following steps (A) and (B): (A) a step of culturing a microorganism able to produce a basic amino acid in a culture medium so that bicarbonate ions and/or carbonate ions serve as counter ions for the basic amino acid, to obtain a fermentation broth containing the basic amino acid; (B) a step of subjecting the fermentation broth to a heat treatment under a pressure sufficient for preventing generation of carbon dioxide gas from the fermentation broth.

## Claims

### Claim 1

A method for producing a basic amino acid or a fermentation product containing the basic amino acid, the method comprising:
(A) culturing a microorganism able to produce a basic amino acid in a culture medium so that bicarbonate ions and/or carbonate ions serve as counter ions for the basic amino acid, to obtain a fermentation broth containing the basic amino acid; and
(B) heat treating the fermentation broth under a pressure sufficient to prevent generation of carbon dioxide gas from the fermentation broth.

### Claim 2 (depends on 1)

The method according to claim 1,
wherein said pressure is gauge pressure and is equal to or higher than the total value of the partial pressure of carbon dioxide and the partial pressure of water vapor at the temperature of said heat treatment, and
wherein said partial pressure of carbon dioxide is determined using the concentrations of the basic amino acid, CO2, NH3, K, Mg, Cl, SO4, acetic acid, and succinic acid in the fermentation broth and the temperature of the heat treating as variables.

### Claim 3 (depends on 1)

The method according to claim 1, wherein said pressure is gauge pressure and is equal to or higher than the pressure shown in the following equation:
Pressure (kPa)=1.99×10−3×T2.54
wherein T is the temperature in centigrade of the heat treating.

### Claim 4 (depends on 1)

The method according to claim 1, wherein said pressure is gauge pressure and is equal to or higher than the pressure of a gas phase in a sealed vessel observed when the fermentation broth is put in the sealed vessel so that the porosity of said vessel becomes 6% v/v and adjusted to the temperature of said heat treating.

### Claim 5 (depends on 1)

The method according to claim 1, wherein said pressure is gauge pressure and is 400 kPa or higher.

### Claim 6 (depends on 1)

The method according to claim 1, wherein said pressure is gauge pressure and is 1000 kPa or higher.

### Claim 7 (depends on 1)

The method according to claim 1, wherein the temperature of the heat treating is 80° C. to 130° C.

### Claim 8 (depends on 1)

The method according to claim 1, wherein the temperature of the heat treating is 100° C. to 130° C.

### Claim 9 (depends on 1)

The method according to claim 1, wherein said heat treating is carried out in a pressure vessel under conditions wherein substantially no gas phase is present in the pressure vessel.

### Claim 10 (depends on 1)

The method according to claim 1, further comprising:
decarboxylating the fermentation broth after said heat treating.

### Claim 11 (depends on 1)

The method according to claim 1, wherein said culturing is carried out while controlling the pH of the culture medium to 7.2 to 9.0 during at least a partial period of culture.

### Claim 12 (depends on 1)

The method according to claim 1, wherein said culturing is carried out so that the pH of the culture medium at the completion of culture is 7.2 or higher.

### Claim 13 (depends on 1)

The method according to claim 1, wherein said culturing is carried out so that bicarbonate ions and/or carbonate ions are present in the culture medium at a concentration of 20 mM or more during at least a partial period of culture by controlling the internal pressure of a fermentation tank to be positive and/or by supplying carbon dioxide gas into the culture medium.

### Claim 14 (depends on 1)

The method according to claim 1, wherein said culturing is carried out so that the concentration of anions other than bicarbonate ions and/or carbonate ions in the culture medium is 900 mM or lower.

### Claim 15 (depends on 13)

The method according to claim 13, wherein the internal gauge pressure of the fermentation tank is 0.03 to 0.2 MPa.

### Claim 16 (depends on 1)

The method according to claim 1, wherein said culturing is carried out so that the total ammonia concentration in the culture medium is controlled to 300 mM or lower during at least a partial period of culture.

### Claim 17 (depends on 1)

The method according to claim 1, wherein the basic amino acid is L-lysine.

### Claim 18 (depends on 1)

The method according to claim 1, wherein the microorganism is a coryneform bacterium or Escherichia coli.

### Claim 19 (depends on 1)

The method according to claim 1, wherein the fermentation product is selected from the group consisting of the fermentation broth, a supernatant of the fermentation broth, a decarboxylation product of the fermentation broth, a concentrated product of the fermentation broth, a dried product of the fermentation broth, and a processed product of the fermentation broth.