Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Method for production of L-lysine

A technology of lysine and diaminopimelate dehydrogenase, which is applied in the direction of biochemical equipment and methods, enzymes, isomerases, etc., can solve the problem of reducing

Active Publication Date: 2014-09-24
AJINOMOTO CO INC
View PDF26 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it was not expected that reducing the activity of enzymes belonging to the meso-DAP synthesis pathway is effective for L-lysine production when DDH is expressed in bacteria of the genus Escherichia

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
  • Method for production of L-lysine
  • Method for production of L-lysine
  • Method for production of L-lysine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0139] [Example 1] Construction of L-lysine-producing bacteria in which dapD is disrupted

[0140] Construction of dapD gene disrupted strain

[0141] First, the dapD-disrupted strain was constructed using the Escherichia coli wild-type strain MG1655.

[0142] Take pMW118(λattL-Km r -λattR) (refer to International Publication No. WO2006093322) plasmid as a template, using SEQ ID NO: 19 and 20, the sequences attL and attR corresponding to the sequence attL and attR at the 3' end of the primer 1. A synthetic oligonucleotide having a sequence corresponding to a part of the dapD gene as the target gene at the 5' end of the primer was used as a primer to carry out PCR, and the λ- MG1655ΔdapD::att-Km strain was constructed by red method. In the λ-red method, Km-resistant recombinants are obtained by plate culture on L-agar medium containing Km (kanamycin) (50mg / L) at 37°C and selecting Km-resistant recombinants. ongoing.

[0143] Transform L-lysine producing strain WC196LC / pC...

Embodiment 2

[0147] [Example 2] Evaluation of L-lysine production capacity of L-lysine-producing bacteria with dapD destroyed

[0148] Melt the glycerol preservation solution of the bacterial strain obtained in Example 1, take 100 μL and evenly spread it on an L plate containing 20 mg / L streptomycin, and incubate at 37° C. for 24 hours. About 1 / 8 of the bacterial cells on the obtained plate were suspended in 0.5 mL of physiological saline, and the turbidity at a wavelength of 600 nm was measured with a spectrophotometer U-2000 (Hitachi). Inoculate 20 mL of the fermentation medium (MS medium, composed as follows) containing 20 mg / L streptomycin in a 500 mL Sakaguchi flask with the resulting suspension containing the bacterium, wherein the amount of liquid inoculated is such that the turbidity at a wavelength of 600 nm was 0.15, and cultured for about 24 hours with a reciprocating shaking culture device at a stirring speed of 114 rpm and a temperature of 37°C. After the cultivation, the amo...

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

L-lysine is produced by culturing the following Escherichia coli having L-lysine-producing ability in a medium modified to reduce the endogenous One or more enzymes of the racemic α,ε-diaminopimelic acid synthesis pathway, such as 2,3,4,5-tetrahydropyridine-2,6-dicarboxylic acid N-succinyltransferase, The activity of succinyldiaminopimelate transaminase, succinyldiaminopimelate desuccinylase or diaminopimelate epimerase, and a gene encoding diaminopimelate dehydrogenase is introduced therein.

Description

technical field [0001] The present invention relates to a method for producing L-lysine using Escherichia coli. L-Lysine is an essential amino acid and is used as a component of nutritional mixtures such as pharmaceuticals or feed supplements for animals. Background technique [0002] L-amino acids such as L-lysine are industrially produced by fermentation using amino acid producing bacteria such as coryneform bacteria or Escherichia bacteria capable of producing these L-amino acids. As these amino acid producing bacteria, strains isolated from nature, artificial mutants of the strains, recombinants in which the activity of L-amino acid biosynthetic enzymes have been enhanced by genetic recombination, etc. have been used in order to increase productivity. The production method of L-lysine includes methods described in Patent Documents 1 to 4, for example. [0003] As a method of improving the productivity of amino acids such as L-lysine, in addition to the method of increa...

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): C12N15/09C12N1/21C12P13/08
CPCC12N9/1096C12P13/08C12N9/90C12N9/1029
Inventor 土井秀高植田拓嗣
Owner AJINOMOTO CO INC
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com