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A method for improving the activity of glutamic acid decarboxylase with 732 cation exchange resin

A technology of glutamic acid decarboxylase and cation exchange, applied in the biological field, can solve the problems of high cost of gamma-aminobutyric acid, etc., and achieve the effects of easy separation and recovery, no corrosion of equipment, and no pollution to the environment

Active Publication Date: 2021-03-02
LINGNAN NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0029] The object of the present invention is to provide a kind of method that improves Enterococcus faecium glutamic acid decarboxylase conversion activity by 732 cation exchange resins, by using 732 cation exchange resins as auxiliary catalyst, 732 cation exchange resins, containing glutamic acid decarboxylase Enterococcus faecium cells, buffers and reaction substrates L - Glutamate or L -Glutamic acid monosodium salt constitutes a co-catalytic system, and finally achieves the purpose of promoting the conversion activity of glutamate decarboxylase in Enterococcus faecium cells, increasing the production of γ-aminobutyric acid, and solving the problem of high cost of biosynthesis of γ-aminobutyric acid

Method used

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  • A method for improving the activity of glutamic acid decarboxylase with 732 cation exchange resin
  • A method for improving the activity of glutamic acid decarboxylase with 732 cation exchange resin
  • A method for improving the activity of glutamic acid decarboxylase with 732 cation exchange resin

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Embodiment 1

[0064] A kind of method utilizing 732 cation exchange resins of the present invention to improve glutamic acid decarboxylase activity comprises the following steps:

[0065] ①. Will L - Dissolve monosodium glutamate (MSG) in 0.2 mol / L acetic acid-sodium acetate buffer solution, adjust the pH to 4.2-4.6, and prepare 0.2 mol / L-0.3 mol / L L - monosodium glutamate solution, as substrate solution;

[0066] ②. Regenerate or pretreat the 732 cation exchange resin with 1 mol / L NaOH and 1 mol / L HCl according to the instructions of the resin, then fully equilibrate it with pH 4.2~pH 4.6, 0.2 mol / L acetic acid-sodium acetate buffer solution, and then Equilibrate with substrate solution;

[0067] ③. The mass of the 732 cation exchange resin balanced according to step ②: the volume of the substrate solution: the volume of the Enterococcus faecium GDMCC60203 bacterial suspension is mixed in a ratio of 1:1:1, and the mixture is the compound conversion reaction of glutamic acid decarboxylase...

Embodiment 2

[0071] A kind of method utilizing 732 cation exchange resins of the present invention to improve glutamic acid decarboxylase activity comprises the following steps:

[0072] ①. Will L - Glutamic acid was dissolved in 0.2 mol / L acetic acid-sodium acetate buffer solution, adjusted to pH 4.2-4.6, and prepared as 0.2 mol / L-0.3 mol / L L - monosodium glutamate solution, as substrate solution;

[0073] ②. Regenerate or pretreat the 732 cation exchange resin with 1 mol / L NaOH and 1 mol / L HCl according to the instructions of the resin, then fully equilibrate it with pH 4.2~pH 4.6, 0.2 mol / L acetic acid-sodium acetate buffer solution, and then Equilibrate with substrate solution;

[0074] ③. The mass of the 732 cation exchange resin balanced according to step ②: the volume of the substrate solution: the volume of the Enterococcus faecium GDMCC60203 bacterial suspension is mixed in a ratio of 1:1:1, and the mixture is the compound conversion reaction of glutamic acid decarboxylase syst...

Embodiment 3

[0078] A kind of method utilizing 732 cation exchange resins of the present invention to improve glutamic acid decarboxylase activity comprises the following steps:

[0079] ①. Will L - Dissolve monosodium glutamate (MSG) in 0.2 mol / L acetic acid-sodium acetate buffer solution, adjust the pH to 4.2-4.6, and prepare 0.2 mol / L-0.3 mol / L L - monosodium glutamate solution, as substrate solution;

[0080] ②. Regenerate or pretreat the 732 cation exchange resin with 1 mol / L NaOH and 1 mol / L HCl according to the instructions of the resin, then fully equilibrate it with pH 4.2~pH 4.6, 0.2 mol / L acetic acid-sodium acetate buffer solution, and then Equilibrate with substrate solution;

[0081] ③. The mass of 732 cation exchange resin balanced according to step ②: the volume of substrate solution: the volume of the free enzyme solution of Enterococcus faecium GDMCC60203 glutamic acid decarboxylase is 1: 1: 1. Mix the three, and the mixture is glutamic acid Decarboxylase complex conver...

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Abstract

The invention relates to a method for improving activity of enterococcus faecium glutamate decarboxylase by virtue of 732 cation exchange resin, and belongs to the field of bio-technology. According to the method provided by the invention, the 732 cation exchange resin is taken as an enzyme activity accelerant of the enterococcus faecium glutamate decarboxylase; a 732 cation exchange resin-glutamate decarboxylase composite catalysis system is constructed by mixing the 732 cation exchange resin, an L-glutamic acid solution and an enterococcus faecium suspension or glutamate decarboxylase free enzyme liquid at the ratio of 1 to 1 to 1 of the mass of the 732 cation exchange resin to the volume of the L-glutamic acid solution to the volume of the enterococcus faecium suspension or glutamate decarboxylase free enzyme liquid; and the yield of [gamma]-aminobutyric acid can be improved by 25.31-143.23% when it conducts a reaction in a water-bath oscillator at 80r / min and 37-43 DEG C or it conducts a stirring reaction at a low speed for 24-36h. According to the method provided by the invention, the 732 cation exchange resin can obviously improve the activity of the glutamate decarboxylase,and meanwhile, an exchange adsorption effect of the 732 cation exchange resin on [gamma]-aminobutyric acid is a process of purifying the [gamma]-aminobutyric acid, so that a downstream extraction andpurification process is simplified and production cost is reduced; and the method is simple and convenient and is green and environment-friendly.

Description

technical field [0001] The invention relates to a method for improving the activity of glutamic acid decarboxylase with 732 cation exchange resin, which belongs to the field of biotechnology. Background technique [0002] γ-Aminobutyric acid (γ-Aminobutyric acid, GABA) is a non-protein amino acid with 4 carbon atoms. It is the main inhibitory neurotransmitter in the mammalian central nervous system. Antioxidant, anti-inflammatory, anti-cancer, blood pressure lowering, sedative and sleep improving effects have become the ingredients of pharmaceuticals and health products that have attracted much attention. GABA is also an important raw material for the production of bioplastic polyamide 4 due to the formation of 2-pyrrolidone by lactamization. [0003] Microbes grow and multiply quickly, and their production of GABA is not limited by time and space. Therefore, the production of GABA by microbial glutamate decarboxylase (glutamate decarboxylase, GAD, EC4.1.1.15) has attracted...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P13/00C12R1/01
Inventor 杨胜远韦锦
Owner LINGNAN NORMAL UNIV
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