Method for increasing activity of GAD (Glutamate Decarboxylase) by using D101 macroporous adsorption resin

A technology of glutamic acid decarboxylase and adsorption resin, which is applied in the biological field and can solve the problems of high cost of gamma-aminobutyric acid

Active Publication Date: 2017-11-07
LINGNAN NORMAL UNIV
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  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0029] The object of the present invention is to provide a method for improving the conversion activity of Enterococcus faecium glutamic acid decarboxylase by D101 macroporous adsorption resin, by using D101 macroporous adsorption resin as auxiliary catalyst, D101 macroporous adsorption resin, containing glutamic acid Decarboxylase Enterococcus fae

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  • Method for increasing activity of GAD (Glutamate Decarboxylase) by using D101 macroporous adsorption resin
  • Method for increasing activity of GAD (Glutamate Decarboxylase) by using D101 macroporous adsorption resin
  • Method for increasing activity of GAD (Glutamate Decarboxylase) by using D101 macroporous adsorption resin

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Embodiment 1

[0064] A kind of method of improving glutamic acid decarboxylase activity with D101 macroporous adsorption resin of the present invention comprises the following steps:

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

[0066] ②. Pretreat the D101 macroporous adsorption resin with 95% ethanol and distilled water according to the instructions of the resin, and then balance it with the substrate solution;

[0067] ③. According to step ②, the mass of D101 macroporous adsorption resin balanced: the volume of substrate solution: the volume of Enterococcus faecium GDMCC 60203 bacterial suspension is mixed in a ratio of 1:1:1, and the mixture is glutamic acid decarboxylase compound Transformation reaction system;

[0068] ④. React the mixture in step ③ in 80r / min, 37℃~43℃ water bath shaker or stir...

Embodiment 2

[0071] A kind of method of improving glutamic acid decarboxylase activity with D101 macroporous adsorption resin of the present invention comprises the following steps:

[0072] ①. Dissolve L-glutamic acid in 0.2mol / L acetic acid-sodium acetate buffer solution, adjust the pH to 4.2-4.6, and prepare a 0.2mol / L-0.3mol / L L-glutamic acid solution as a substrate solution;

[0073] ②. Pretreat the D101 macroporous adsorption resin with 95% ethanol and distilled water according to the instructions of the resin, and then balance it with the substrate solution;

[0074] ③. According to step ②, the mass of D101 macroporous adsorption resin balanced: the volume of substrate solution: the volume of Enterococcus faecium GDMCC 60203 bacterial suspension is mixed in a ratio of 1:1:1, and the mixture is glutamic acid decarboxylase compound Transformation reaction system;

[0075] ④. React the mixture in step ③ in 80r / min, 37℃~43℃ water bath shaker or stir at 37℃~43℃ at low speed or stand fo...

Embodiment 3

[0078] A kind of method utilizing D101 macroporous adsorption resin of the present invention to improve glutamic acid decarboxylase activity comprises the following steps:

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

[0080] ②. Pretreat the D101 macroporous adsorption resin with 95% ethanol and distilled water according to the instructions of the resin, and then balance it with the substrate solution;

[0081] ③. The mass of D101 macroporous adsorption resin balanced according to step ②: volume of substrate solution: volume of Enterococcus faecium GDMCC 60203 glutamate decarboxylase free enzyme solution is 1:1:1 and the three are mixed, and the mixture is gluten Amino acid decarboxylase complex transformation reaction system;

[0082] ④. React the mixture in step ③ in 80r / min, 37℃~43...

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Abstract

The invention relates to a method for increasing the activity of enterococcus faecium Glutamate Decarboxylase by using D101 macroporous adsorption resin, and belongs to the technical field of biology. According to the method disclosed by the invention, the D101 microporous adsorption resin is used as an enzyme activity accelerator of enterococcus faecium GAD, and a D101 microporous adsorption resin-GAD composite catalytic system is constructed according to a proportion that the mass of the D101 macroporous adsorption resin to the volume of a substrate solution to the volume of an enterococcus faecium bacterial suspension solution or a GAD free enzyme solution is 1 to 1 to 1; when the D101 microporous adsorption resin-GAD composite catalytic system is subjected to reaction in a water bath oscillator of which the temperature is 37 to 43 DEG C at 80r/min or stirring reaction in a stirring tank at a low speed for 24 to 36 hours, the yield of GABA (Gamma-Aminobutyric Acid) can be increased by 18.54 to 149.11 percent; the D101 macroporous adsorption resin is capable of remarkably increasing the activity of the GAD, an exchange adsorption function of the D101 microporous adsorption resin on the GABA is also a purifying process of the GABA, a downstream extracting and purifying technology is simplified, and the production cost is reduced; the method is simple, green and environment-friendly.

Description

technical field [0001] The invention relates to a method for improving glutamic acid decarboxylase activity by using D101 macroporous adsorption 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 attr...

Claims

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Application Information

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IPC IPC(8): C12P13/00C12R1/01
CPCC12P13/005
Inventor 杨胜远韦锦
Owner LINGNAN NORMAL UNIV
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