An L-amino acid-producing bacterium and a method for producing an L-amino acid

An amino acid and leucine technology, which is applied in microorganism-based methods, biochemical equipment and methods, botanical equipment and methods, etc., can solve the problem of unknown mutation of yeaS gene, and achieve the effect of high-efficiency fermentation production

Active Publication Date: 2014-12-24
AJINOMOTO CO INC
View PDF134 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] As mentioned above, by increasing the expression level of the yeaS gene, its effect on the production of various amino acids can be studied (Non-Patent Document 2), but the mutation of the yeaS gene that improves the productivity of L-amino acid is still unknown

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • An L-amino acid-producing bacterium and a method for producing an L-amino acid
  • An L-amino acid-producing bacterium and a method for producing an L-amino acid
  • An L-amino acid-producing bacterium and a method for producing an L-amino acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0242] (Example 1) Effect of increasing L-cysteine ​​production by strengthening wild-type YeaS

[0243] In order to study the effect on L-cysteine ​​production due to enhanced expression of the wild-type yeaS gene in P. ananatis, a gene cysE5 encoding a mutant SAT was introduced (US Patent Application No. 20050112731), and A strain having an increased copy number of the yeaS gene.

[0244] First, the plasmids used to construct the above strains were constructed. Its method is as follows.

[0245] Using the chromosomal DNA of Escherichia coli MG1655 (ATCC No.47076) as a template, using P1 (agctgagtcg acccccagga aaaattggtt aataac: SEQ ID NO: 20) and P2 (agctgagcat gcttccaact gcgctaatga cgc: SEQ ID NO: 21) as primers, obtained by PCR A DNA fragment of about 300 bp containing the promoter region of the nlpD gene (hereinafter, the wild-type nlpD gene promoter is referred to as "Pnlp0") was obtained. Sites for restriction endonucleases SalI and PaeI were designed at the 5' end a...

Embodiment 2

[0259] (Example 2) Obtaining the mutant yeaS gene and L-cysteine ​​production increasing effect

[0260] Next, random mutations were artificially introduced into the yeaS gene by error-prone PCR in order to obtain a mutant with a higher activity of increasing the L-cysteine ​​production capacity of the host cell than the wild-type YeaS.

[0261] (1) Preparation of error-prone PCR and mutation introduction library of yeaS

[0262] First, the error-prone PCR conditions for introducing 1 to 3 mutations into the yeaS gene were based on existing knowledge (Evert Bokma et al., (2006) FEBS Letters 580:5339-5343, Zao et al., (1998) Nat Biotechnol Mar; 16(3):258-61). DNA polymerase uses taq polymerase (manufactured by QIAGEN), and the composition of the reaction solution is 10mM Tris-HCl (pH8.3), 50mM KCl, 7mM MgCl 2 , 0.2mM dGTP, dATP, 1mM dCTP, dTTP and 12.5μM MnCl 2 . The primers used were P9 (catgccatgg tcgctgaata cggggttctg, SEQ ID NO: 28) and P10 (aactgcagtc aggattgcag cgtcgc...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

Bacteria belonging to the family Enterobacteriaceae that have the ability to produce amino acids such as L-cysteine ​​and have been modified to carry a yeaS gene with a specific mutation are cultivated in the medium, so that L-amino acids are generated and accumulated in the medium, and from This medium collects L-amino acids, thereby producing L-amino acids.

Description

technical field [0001] The present invention relates to a method for producing L-amino acids such as L-cysteine, and more specifically, to a bacterium suitable for producing L-amino acid, and a method for producing L-amino acid using the bacterium. L-amino acids are used in various fields such as seasonings, food additives, feed additives, chemical products, and pharmaceuticals. Background technique [0002] L-amino acids are industrially produced by fermentation methods using microorganisms belonging to the genus Brevibacterium, Corynebacterium, Escherichia, and the like. In these production methods, strains isolated from nature or artificial mutants of the strains are used, and microorganisms modified by recombinant DNA technology to increase the activity of basic L-amino acid biosynthetic enzymes, etc. are used (Patent Documents 1 to 10). 9). [0003] In addition, for example, L-cysteine ​​can be obtained by extraction from keratin-containing substances such as hair, ho...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Patents(China)
IPC IPC(8): C12P13/04C12P13/12C12P13/06C12P13/08C12P13/24C12P13/14C12P13/10C12P13/22C12N1/21C12N15/31C12R1/01C12R1/19
CPCC07K14/245C12P13/12
Inventor 宅见和浩野中源
Owner AJINOMOTO CO INC
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap