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

A gene encoding 6-phosphogluconate dehydrogenase and its application

A technology of phosphoglucose and acid dehydrogenase, which is applied in the field of genetic engineering and can solve problems such as low yield

Active Publication Date: 2019-03-15
GUANGXI UNIV
View PDF5 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Aiming at the problems of feedback inhibition regulation and relatively low yield of L-serine produced by the fermentation of Methylobacterium sp.MB200, the present invention utilizes plasmid transposon insertion technology to construct a mutant library of Methylobacterium sp.MB200, A mutant with D-serine resistance was screened out, the 6-phosphogluconate dehydrogenase (6-PGDH) gene with L-serine inhibitory effect was cloned, and a pgdh gene-deficient L-serine high-yielding strain was constructed , relieve the inhibitory effect of 6-PGDH on the production of L-serine during the metabolism of methylotrophic bacteria, thereby increasing the production of L-serine

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
  • A gene encoding 6-phosphogluconate dehydrogenase and its application
  • A gene encoding 6-phosphogluconate dehydrogenase and its application
  • A gene encoding 6-phosphogluconate dehydrogenase and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] 1.1 Cloning and analysis of the D-serine tolerance mutant gene of methylobacterium Methylobacterium sp.MB200

[0034] 1.1.1 A mutant library of methylobacterium Methylobacterium sp.MB200 was constructed by plasmid transposon insertion technology, and the mutant D-2 with D-serine resistance was screened out.

[0035] 1.1.2 Extract the total DNA from the mutant D-2, connect it to the cloning vector pMD-18T after enzyme digestion, and introduce it into the competent E. Plasmids were further sequenced after digestion and verification, and Blast sequence alignment and analysis were performed in NCBI. As a result, the obtained DNA fragment was 1875bp, and the sequence had the highest similarity with the 6-phosphogluconate dehydrogenase gene of Methylobacterium populi BJ001, with a consistency of 89%. See Table 1 for details.

[0036] Mutant DNA fragments cloned in table 1

[0037]

[0038] 1.2 Bioinformatics analysis of inserted DNA fragments in mutant strain D-2

[003...

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

The invention provides a gene of coded 6-phosphogluconate dehydrogenase and application thereof. The nucleotide sequence is shown in SEQ ID NO:1. The 6-phosphogluconate dehydrogenase gene pgdhD2 is derived from Methylobacterium sp. MB200 and has the L-serine inhibiting effect. The Methylobacterium sp. MB200 is used as an original strain, pgdh gene deletion mutation DMB is built through a homologous double-crossover method, an L-serine high-yield mutant strain with the L-serine producing capacity greatly higher than that of original strain Methylobacterium sp. MB200 is obtained, and the tolerated D-serine concentration of the L-serine high-yield mutant strain is greatly higher than that of the original strain. The pgdh gene deletion mutation stain has the good application prospect in production of L-serine through a fermentation method.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, in particular to a gene encoding 6-phosphogluconate dehydrogenase and its application. Background technique [0002] L-serine has important physiological functions and application value, and is widely used in the fields of medicine, food and chemical industry. At present, the production methods of L-serine mainly include chemical synthesis, proteolysis, enzymatic transformation and microbial fermentation. The production of L-serine by microbial fermentation is a green and environmentally friendly production method, but due to the feedback inhibition regulation, it is difficult for microorganisms to accumulate L-serine in large quantities, so the yield of L-serine produced by microbial fermentation is relatively low. application of this method. [0003] The method for removing the feedback inhibition regulation of L-serine and improving the yield of L-serine produced by fermentation ...

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): C12N15/53C12N9/04C12N15/70
CPCC12N9/0006
Inventor 申佩弘武波李献蒋承建
Owner GUANGXI UNIV
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