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Method for catalytically synthesizing gamma-aminobutyric acid by using sodium glutamate and immobilized bio-enzyme

A technology of aminobutyric acid and sodium glutamate, applied in directions such as fixing on/in organic carriers, fermentation, etc., can solve the safety problems of Escherichia coli and other problems, achieve short cycle, specific reaction and high yield Effect

Inactive Publication Date: 2012-09-19
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

AI Technical Summary

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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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Weigh 18.7g sodium glutamate (monohydrate) and add 1000ml purified water to dissolve, adjust pH=5.5 with 1:1 hydrochloric acid diluent, add 10kU solid-phase enzyme to make the concentration of sodium glutamate 100mmols / L, heat in water bath, 37 ℃ heat preservation and stirring reaction, continuously add 1:1 hydrochloric acid diluent to maintain the pH of the reaction solution = 5.5. Samples were taken every 30 minutes to detect the content of sodium glutamate to determine the conversion rate of the reaction; when the conversion rate of γ-aminobutyric acid reached 99%, the reaction solution was separated from the solid-phase enzyme by filtration.

[0061] Slowly flow the filtrate through a strong acidic ion exchange resin (resin equivalent: 0.21 mol), and then elute with pure water. After washing to neutrality, elute with 2M dilute ammonia water. The amount of ammonia water is based on resin equivalent.

[0062] Add 0.5 g of activated carbon to the ammonia water eluent, ...

Embodiment 2

[0065] Weigh 93.5g of sodium glutamate (monohydrate), add 500ml of water and 50kU of solid-phase enzyme and dilute to 1000ml with pure water, so that the concentration of sodium glutamate is 500mmols / L. Heat in a water bath, keep stirring at 37°C for reaction, and continuously add 1:1 diluent of hydrochloric acid to maintain the pH of the reaction solution at 5.5. Samples were taken every 30 minutes to detect the content of sodium glutamate to determine the conversion rate of the reaction; when the conversion rate of γ-aminobutyric acid reached 99%, the reaction solution was separated from the solid-phase enzyme by filtration.

[0066] Slowly flow the filtrate through a strong acidic ion exchange resin (resin equivalent: 1.1 mol), and then elute with pure water until neutral, then elute with 2M dilute ammonia.

[0067] Add 2.0 g of activated carbon to the ammonia water eluent, stir for 30 minutes, and decarburize by filtering through a 0.22 μm filter membrane. The filtrate is...

Embodiment 3

[0070] Weigh 1000mmols sodium glutamate (monohydrate) 187.0g, add water 500ml and 100kU solid-phase enzyme, dilute to 1000ml with pure water, so that the concentration of sodium glutamate is 1000mmols / L. Heat in a water bath, keep stirring at 37°C, and keep adding 1:1 diluent of hydrochloric acid to maintain the pH of the reaction solution at 5.5. Samples were taken every 30 minutes to detect the content of sodium glutamate to determine the conversion rate of the reaction; when the conversion rate of γ-aminobutyric acid reached 99%, the reaction solution was separated from the solid-phase enzyme by filtration.

[0071] The filtrate was slowly flowed through a strong acidic ion exchange resin (resin equivalent: 2.1 mol), and then eluted with pure water. After washing to neutrality, it was eluted with 2M dilute ammonia water.

[0072] Add 4.0 g of activated carbon to the ammonia water eluent, stir for 30 minutes, and decarburize by filtering through a 0.22 μm filter membrane. T...

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Abstract

The invention provides a method for catalytically synthesizing gamma-aminobutyric acid by using sodium glutamate and an immobilized bio-enzyme. The main process comprises the following steps: (1) performing catalytic synthesis on a substrate, namely a sodium glutamate solution and an immobilized enzyme to obtain gamma-aminobutyric acid, and performing centrifugal separation on the immobilized bio-enzyme from a reaction liquid; (2) allowing the reaction liquid to pass through a cation exchange resin to remove sodium chloride from the reaction liquid so as to obtain purified gamma-aminobutyric acid; (3) decoloring the aqueous solution of gamma-aminobutyric acid by using active carbon, and concentrating by using a vacuum membrane to obtain a concentrated solution of gamma-aminobutyric acid; and (4) adding 95 percent alcohol into the concentrated solution of gamma-aminobutyric acid, separating out white gamma-aminobutyric acid precipitate crystals, performing centrifugal separation and vacuum drying to obtain white gamma-aminobutyric acid powder. The process has the advantages of reaction specificity, high yield, high 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 sodium glutamate biological solid-phase enzyme, which belongs to the 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 exists almost only in nerve tissue, and the content in brain tissue is about 0.1-0.6mg / g 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 has been studied...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/00C12N11/08
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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