Method for producing l-cysteine

A technology of cysteine ​​and production capacity, applied in the field of producing L-cysteine ​​or its related substances, can solve the problem of not suggesting cysG gene and the like

Inactive Publication Date: 2014-11-19
AJINOMOTO CO INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Furthermore, none of the above findings involving amino acid production suggest that enhanced expression of the cysG gene, cysJ gene, or cysI gene is effective for amino acid production using thiosulfuric acid as a sulfur source

Method used

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  • Method for producing l-cysteine
  • Method for producing l-cysteine
  • Method for producing l-cysteine

Examples

Experimental program
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Effect test

Embodiment 1

[0175] Example 1: Production of cysteine ​​by Escherichia coli overexpressing the cysG or cysGJI gene using thiosulfuric acid as a sulfur source

[0176] (1) Construction of expression plasmids of genes involved in sulfurous acid reduction

[0177] As for genes involved in sulfite reduction, the cysG gene of Pantoea ananatis SC17 strain, the cysGJI gene of Pantoea ananatis SC17 strain, the cysG gene of Escherichia coli M1655 strain, and the cysGJI gene of Escherichia coli MG1655 strain were used. Each of these genes was cloned into an expression vector to construct an expression plasmid for the gene. The steps are described in (1-1) to (1-7).

[0178] (1-1) Construction of expression vector

[0179] As the expression vector, pMIV-Pnlp23-ter (Japanese Patent Laid-Open No. 2008-99668 ) constructed from pMIV-5JS was used. This vector has a strong promoter nlp23 promoter (Pnlp23) and rrnB terminator, and has SalI and XbaI sites between Pnlp23 and rrnB terminator. By using prim...

Embodiment 2

[0249] Example 2: Production of cysteine ​​by Pantoea ananatis overexpressing the cysG gene or the cysGJI gene using thiosulfate as a sulfur source

[0250] (1) Construction of Pantoea ananatis cysteine ​​producing strain

[0251] Regarding the Pantoea ananatis cysteine-producing strain, an EYPSint-1MΔd0191(S) strain was constructed in which the cysE5 gene, the yeaS gene, the serA348 gene, and the cysM gene were introduced into the chromosome, the expression of the cysPTWA gene was enhanced in the strain, and d0191 gene deletion. The construction steps are shown in (1-1) to (1-5).

[0252] (1-1) Introducing cysE5 gene and yeaS gene into Pantoea ananatis SC17 strain

[0253] The cysE5 gene and the yeaS gene were introduced into the chromosome of the Pantoea ananatis SC17 strain to construct the EY19(s) strain. The cysE5 gene is a mutant gene of the cysE gene, which encodes a mutant serine acyltransferase (SAT), in which the Val residue and Asp residue at positions 95 and 96 ...

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Abstract

A method for producing L-cysteine and the like is provided by developing a novel technique for improving bacterial L-cysteine-producing ability. By culturing a bacterium belonging to the family Enterobacteriaceae and having L-cysteine-producing ability, of which expression of a gene involved in sulfite reduction is enhanced, in a medium containing thiosulfate and collecting L-cysteine, a related substance thereof, or a mixture thereof accumulated in the medium, these compounds are produced.

Description

field of invention [0001] The present invention relates to a method for producing L-cysteine ​​or its related substances. L-cysteine ​​and its related substances are used in the fields of medicine, beauty and food. Background technique [0002] Conventionally, L-cysteine ​​is extracted from keratin-containing substances such as hair, horn, and feather, or converted into DL-2-aminothiazoline-4-carboxylic acid (DL-2 -aminothiazoline-4-carboxylic acid) obtained. There are also plans to use the new enzyme to produce L-cysteine ​​on a large scale by the immobilized enzyme method. Furthermore, there have been attempts to produce L-cysteine ​​by fermentation using microorganisms. [0003] The L-cysteine ​​productivity of microorganisms can be improved by enhancing the activity of enzymes in the L-cysteine ​​biosynthetic pathway, or enhancing the activity of enzymes involved in the production of substrates (such as L-serine) of the pathway . As microorganisms having the ability...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/09C12P13/12C12R1/01C12R1/19
CPCC12N9/1007C12N9/0051C12Y108/01002C12Y201/01107C12P13/12
Inventor 山崎俊介野中源宅见和浩
Owner AJINOMOTO CO INC
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