# US 2013/0157321 A1 · Use of Phosphoketolase for Producing Useful Metabolites

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- Number: US 2013/0157321 A1
- Document ID: US20130157321A1
- Status: Published application
- Kind: A1
- Application number: 13/774,153
- Filed: 2013-02-22
- Published: 2013-06-20
- Earliest claimed: 2004-08-10
- Assignees: Ajinomoto Co., Inc.
- Inventors: Yury Ivanovich Kozlov; Akito Chinen; Hiroshi Izui; Yoshihiko Hara; Hisashi Yasueda; Konstantin Vyacheslavovich Rybak; Ekaterina Aleksandrovna Slivinskaya; Joanna Yosifovna Katashkina
- Web page: https://patentgrep.com/patent/US20130157321A1

## CPC

- C12P 13/24: Proline; Hydroxyproline; Histidine
- C12N 9/88: Lyases (4.)
- C12P 7/42: Hydroxy-carboxylic acids
- C12P 7/44: Polycarboxylic acids
- C12P 7/46: Dicarboxylic acids having four or less carbon atoms, e.g. fumaric acid, maleic acid
- C12P 7/62: Carboxylic acid esters
- C12P 13/06: Alanine; Leucine; Isoleucine; Serine; Homoserine
- C12P 13/10: Citrulline; Arginine; Ornithine
- C12P 13/12: Methionine; Cysteine; Cystine
- C12P 13/14: Glutamic acid; Glutamine
- C12P 13/20: Aspartic acid; Asparagine
- C12Y 401/02009: Phosphoketolase (4.1.2.9)
- C12Y 401/02022: Fructose-6-phosphate phosphoketolase (4.1.2.22)
- 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

## Abstract

The present invention provides a bacterium which has an ability to produce a useful metabolite derived from acetyl-coenzyme A, such as L-glutamic acid, L-glutamine, L-proline, L-arginine, L-leucine, L-cysteine, succinate, and polyhydroxybutyrate, wherein said bacterium is modified so that activities of D-xylulose-5-phosphate phosphoketolase and/or fructose-6-phosphate phosphoketolase are enhanced. The present invention also provides a method for producing the useful metabolite using the bacterium.

## Claims

### Claim 1

A method for producing a useful metabolite derived from acetyl-coenzyme A comprising cultivating a bacterium in a culture medium to produce a useful metabolite derived from acetyl-coenzyme A in the culture medium and/or the bacterium, and collecting said useful metabolite from the culture medium and/or the bacterium, wherein said bacterium is modified to have an increased activity of D-xylulose-5-phosphate phosphoketolase and/or fructose-6-phosphate phosphoketolase.

### Claim 2 (depends on 1)

The method according to claim 1, wherein the useful metabolite is selected from the group consisting of L-glutamic acid, L-glutamine, L-proline, L-arginine, L-leucine, L-cysteine, succinate, and polyhydroxybutyrate.

### Claim 3 (depends on 1)

The method according to claim 1, wherein the bacterium inherently does not have an activity of D-xylulose-5-phosphate phosphoketolase and fructose-6-phosphate phosphoketolase, and wherein said bacterium is transformed with a DNA fragment coding for D-xylulose-5-phosphate phosphoketolase and/or fructose-6-phosphate phosphoketolase.

### Claim 4 (depends on 1)

The method according to claim 1, wherein the bacterium is further modified to have a reduced activity of 6-phosphofructokinase.

### Claim 5 (depends on 1)

The method according to claim 1, wherein the bacterium is selected from the group consisting of Enterobacteriaceae, coryneform bacterium, and Bacillus bacterium.

### Claim 6 (depends on 1)

The method according to claim 1, wherein the bacterium belongs to the genus Escherichia or Pantoea.

### Claim 7 (depends on 1)

The method according to claim 1, wherein the bacterium is Escherichia coli or Pantoea ananatis.