Variant gene of glutamate decarboxylase and purpose thereof

A glutamic acid decarboxylase, glutamic acid technology, applied in the field of molecular biology, can solve the problem of low catalytic activity

Inactive Publication Date: 2010-12-15
ZHEJIANG UNIV
View PDF3 Cites 13 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The invention provides a variant gene of glutamic acid decarboxylase, which solves the problem of

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
  • Variant gene of glutamate decarboxylase and purpose thereof
  • Variant gene of glutamate decarboxylase and purpose thereof
  • Variant gene of glutamate decarboxylase and purpose thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0017] Construction of recombinant plasmids

[0018] Lactobacillus brevis (Lactobacillus brevis) CGMCC NO.1306 was cultured to the mid-exponential growth stage, and 4.5ml of the bacterial liquid was centrifuged at 10,000rpm for one minute, then the supernatant was discarded, and genomic DNA was extracted using a kit (Shanghai Sangong) according to the instructions.

[0019] Design the following degenerate primers:

[0020] Upstream degenerate primer 1: 5'-cg ggatcc atgaaYaaRaaYgaYcaRgaRc-3';

[0021] Upstream degenerate primer 2: 5'-cg ggatcc atggcWatgttRtaYggWaaac-3';

[0022] and a downstream degenerate primer:

[0023] 5'-gg gaattc ttagtgHgtgaaYccgtattt-3'

[0024] Among them, the " ggatcc "is the BamH I restriction site, in the downstream primer" gaattc” It is the restriction site of EcoR I.

[0025] Use upstream primer 1 (Up1) and upstream primer 2 (Up2) to pair with downstream primer (Down1) respectively, select and optimize different template / Pfu / primer ra...

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 discloses a variant gene of glutamate decarboxylase. DNA sequence of the gene is SEQ ID: No.1. By site-directed mutagenesis on the basis of coded wild GAD enzyme gene, the variant gene according to the invention enhances catalytic activity of the corresponding variant enzyme thereof in neutral pH value and widens the most proper scope of pH value of enzyme, so that the GABA generation ability of the corresponding variant enzyme through the catalysis of glutamic acid on condition of pH=6.0 is twice as much as that of wild-type enzyme. As the most proper scope of pH value of enzyme is widened, catalysis reaction can be conducted on condition that pH is higher than 5.0, thus the problem that the solubility of substrate, i.e. glutamic acid, is low on condition of pH from 4.0 to 5.0 is solved, and conditions are created for GABA production by biotransformation of enzyme.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a variant gene of glutamic acid decarboxylase and its application. Background technique [0002] Site-directed mutagenesis (site-directed mutagenesis or oligonucleotide-directed mutagenesis or site-specific mutagenesis) refers to the technique of introducing specific base pair changes at designated sites of the target DNA fragment. It is a powerful tool for studying the complex relationship between protein structure and function, and it is also a common means of modifying genes in the laboratory. Site-specific changes, deletions, or insertions of specific bases in a known gene can change the corresponding amino acid sequence and the structural and functional characteristics of the protein, and help to understand the relationship between protein structure and function. Especially in the rational design of enzymes, or the use of random mutation and other directed evoluti...

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
IPC IPC(8): C12N15/60C12N9/88C12N15/63C12N1/15C12N1/19C12N1/21C12N5/10C12P13/00
Inventor 梅乐和郁凯胡升黄俊姚善泾
Owner ZHEJIANG UNIV
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