# US 10,501,763 B2 · Microorganism producing O-acetyl-homoserine and method for producing O-acetylhomoserine using the same

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- Number: US 10,501,763 B2
- Document ID: US10501763B2
- Status: Granted
- Kind: B2
- Application number: 15/316,475
- Filed: 2015-06-05
- Published: 2017-05-18
- Granted: 2019-12-10
- Earliest claimed: 2014-06-05
- Assignees: Cj Cheiljedang Corporation
- Applicants: Cj Cheiljedang Corporation
- Inventors: Jee Yeon Bae; Hyun Ah Kim; Yong Uk Shin; So Young Kim; Sang Kyoum Kim; Kwang Ho Na; Ju Hee Seo; Sung Kwang Son; Hye Ryun Yoo; Jin Geun Choi
- Web page: https://patentgrep.com/patent/US10501763B2

## CPC

- C12P 13/06: Alanine; Leucine; Isoleucine; Serine; Homoserine
- C07K 14/245: Escherichia (G)
- C12N 9/0008: {acting on the aldehyde or oxo group of donors (1.2)}
- C12N 9/1029: {transferring groups other than amino-acyl groups (2.3.1)}
- C12N 9/1096: {transferring nitrogenous groups (2.6)}
- C12N 9/1205: {Phosphotransferases with an alcohol group as acceptor (2.7.1), e.g. protein kinases}
- C12N 9/1217: {Phosphotransferases with a carboxyl group as acceptor (2.7.2)}
- C12N 9/88: Lyases (4.)
- C12N 15/52: Genes encoding for enzymes or proenzymes
- C12P 13/02: Amides, e.g. chloramphenicol {or polyamides; Imides or polyimides; Urethanes, i.e. compounds comprising N-C=O structural element or polyurethanes (peptides C12P21/00 or C07K)}
- C12Y 102/01011: Aspartate-semialdehyde dehydrogenase (1.2.1.11)
- C12Y 203/01031: Homoserine O-acetyltransferase (2.3.1.31)
- C12Y 206/01001: Aspartate transaminase (2.6.1.1), i.e. aspartate-aminotransferase
- C12Y 207/01039: Homoserine kinase (2.7.1.39)
- C12Y 207/02004: Aspartate kinase (2.7.2.4)
- C12Y 401/01031: Phosphoenolpyruvate carboxylase (4.1.1.31)
- C12Y 402/01022: Cystathionine beta-synthase (4.2.1.22)
- C12N 1/205: Bacterial isolates
- C12R 2001/12: Bacillus polymyxa {; Paenibacillus polymyxa}
- 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)}
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12Y: ENZYMES
- C12R: INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS

## Abstract

The present disclosure relates to a microorganism producing O-acetylhomoserine with high efficiency and a method for producing O-acetylhomoserine and L-methionine using the microorganism. The present disclosure provides a microorganism producing O-acetylhomoserine having an enhanced activity of a protein which is predicted to export O-acetylhomoserine, and a method for producing O-acetylhomoserine and L-methionine using the microorganism.

## Claims

### Claim 1

A modified microorganism of the genus Escherichia producing O-acetylhomoserine, wherein an activity of the protein comprising the amino acid sequence of SEQ ID NO: 1 in a microorganism of the genus Escherichia capable of producing O-acetylhomoserine is enhanced compared to a non-modified microorganism;
wherein an activity of cystathionine synthase is weakened or inactivated compared to a non-modified microorganism; and
wherein an activity of homoserine acetyltransferase is further enhanced compared to a non-modified microorganism.

### Claim 2 (depends on 1)

The modified microorganism of the genus Escherichia according to claim 1, wherein the microorganism is Escherichia coli.

### Claim 3 (depends on 1)

The modified microorganism of the genus Escherichia according to claim 1, wherein an activity of homoserine kinase is further weakened or inactivated.

### Claim 4 (depends on 1)

The modified microorganism of the genus Escherichia according to claim 1, wherein the activity of at least one enzyme selected from the group consisting of phosphoenolpyruvate carboxylase, aspartate aminotransferase, and aspartate semialdehyde dehydrogenase is further enhanced.

### Claim 5 (depends on 1)

The modified microorganism of the genus Escherichia according to claim 1, wherein an activity of aspartate kinase is enhanced compared to a non-modified microorganism.

### Claim 6 (depends on 1)

A method for producing O-acetylhomoserine comprising culturing the modified microorganism of the genus Escherichia producing O-acetylhomoserine of claim 1 to obtain a cultured medium.

### Claim 7 (depends on 6)

The method according to claim 6, further comprising recovering O-acetylhomoserine from the cultured microorganism or the cultured medium.

### Claim 8 (depends on 1)

A method for producing L-methionine, comprising:
culturing a modified microorganism of the genus Escherichia producing O-acetylhomoserine of claim 1 thereby accumulating O-acetylhomoserine; and
producing L-methionine from the accumulated O-acetylhomoserine by using an enzyme reaction.