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Method for catalytically synthesizing gamma-aminobutyric acid from glutamate biological solid-phase enzyme

A technology of aminobutyric acid and biosynthesis, applied in biochemical equipment and methods, botany equipment and methods, plant genetic improvement, etc., can solve problems such as the safety of Escherichia coli, achieve short cycle, low energy consumption, and high reaction single-minded effect

Inactive Publication Date: 2012-09-26
广东乐尔康生物科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, if it is to be used for food development, there are undoubtedly various problems in terms of safety when using E. coli

Method used

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  • Method for catalytically synthesizing gamma-aminobutyric acid from glutamate biological solid-phase enzyme

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Weigh 14.7g of glutamic acid, add 1000ml of purified water and 10kU of solid-phase enzyme to make the substrate concentration 100mmols / L, heat in a water bath, and keep stirring at 37°C for reaction. Samples were taken every 30 minutes to detect the content of glutamic acid to determine the conversion rate of the reaction; when the conversion rate of γ-aminobutyric acid reached 99% (usually 180-240 minutes), the reaction solution was separated from the solid-phase enzyme by filtration.

[0059] Add 0.5 g of activated carbon to the filtrate of the reaction liquid, stir for 30 minutes, and decarburize by filtering through a 0.22 μm filter membrane. The filtrate was transferred to a rotary thin-film concentrator, and concentrated in vacuum at 60°C until the content of γ-aminobutyric acid was 230-240g / 100ml.

[0060] Add 100ml of 95% alcohol, the white precipitate of γ-aminobutyric acid precipitates, stir and cool to room temperature, filter with suction, wash with 95% alco...

Embodiment 2

[0062] Weigh 73.5g of glutamic acid, add 500ml of water and dilute 50kU of solid-phase enzyme to 1000ml with pure water to make the substrate concentration 500mmols / L, heat in a water bath, and keep stirring at 37°C for reaction. Samples were taken every 30 minutes to detect the content of glutamic acid to determine the conversion rate of the reaction; when the conversion rate of γ-aminobutyric acid reached 99% (usually 180-240 minutes), the reaction solution was separated from the solid-phase enzyme by filtration.

[0063] Add 2.0 g of activated carbon to the filtrate of the reaction liquid, stir for 30 minutes, and decarburize by filtering through a 0.22 μm filter membrane. Transfer the filtrate to a rotary thin-film concentrator; concentrate in vacuum at 60°C until the content of γ-aminobutyric acid is 230-240g / 100ml.

[0064] Add 250 ml of 95% alcohol, and white precipitate of γ-aminobutyric acid precipitates out. The filtrate is stirred and cooled to room temperature, fil...

Embodiment 3

[0066] Weigh 147.0g of glutamic acid, add 500ml of water and dilute 50kU of solid-phase enzyme to 1000ml with pure water, so that the substrate concentration is 1000mmols / L, heat in a water bath, and keep stirring at 37°C for reaction. Samples were taken every 30 minutes to detect the content of glutamic acid to determine the conversion rate of the reaction; when the conversion rate of γ-aminobutyric acid reached 99% (usually 180-240 minutes), the reaction solution was separated from the solid-phase enzyme by filtration.

[0067] Add 5.0 g of activated carbon to the filtrate of the reaction liquid, stir for 30 minutes, and decarburize by filtering through a 0.22 μm filter membrane. The filtrate is transferred to a rotary thin-film concentrator; concentrated in vacuum at 60°C until the content of γ-aminobutyric acid is 230-240g / 100ml.

[0068] Add 1000 ml of 95% alcohol, and white precipitate of γ-aminobutyric acid precipitates out. The filtrate is stirred and cooled to room te...

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Abstract

The invention provides a method for catalytically synthesizing gamma-aminobutyric acid from a glutamate biological solid-phase enzyme, The method comprises the following steps: (1) catalytically synthesizing substrate glutamic acid solution with solid-phase enzyme to generate gamma-aminobutyric acid solution, and then filtering or centrifugally separating the biological solid-phase enzyme from reaction solution; (2) after the activated carbon fading of the gamma-aminobutyric acid solution, concentrating the obtained product in vacuum by a thin film to obtain concentrated gamma-aminobutyric acid solution; and (3) adding 95% alcohol in the gamma-aminobutyric acid solution, crystallizing, centrifugally separating and drying the precipitated gamma-aminobutyric acid white precipitate in vacuum to obtain white gamma-aminobutyric acid powder. The process has the advantages of special reaction, high yield and purity, short period and low energy consumption and is suitable for industrial production.

Description

technical field [0001] The invention relates to a method for synthesizing gamma-aminobutyric acid catalyzed by glutamic acid biological solid-phase enzyme, which belongs to the technical field of biosynthesis. Background technique [0002] GABA Review: [0003] Introduction: [0004] γ-aminobutyric acid, English name: γ-aminobutyric acid (GABA) γ-aminobutyric acid, chemical name: 4-aminobutyric acid, aliases: γ-aminobutyric acid, aminobutyric acid, pipecolic acid. Molecular formula: C 4 h 9 NO 2 . Molecular weight: 103.1. Widely distributed in animals and plants. Seeds, rhizomes and tissue fluids of plants such as beans, ginseng, and Chinese herbal medicines all contain GABA. In animals, GABA is almost only present in nerve tissue, and the content in brain tissue is about 0.1-0.6 mg / gram tissue. Immunological studies have shown that the region with the highest concentration is the substantia nigra in the brain. GABA is an important inhibitory neurotransmitter that h...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/00C12N15/60C12N15/70C12N11/08
Inventor 范永军周丙午周兴华
Owner 广东乐尔康生物科技股份有限公司
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