Production process for synthetizing L-theanine through biological immobilized enzyme catalysis

A production process and biosynthesis technology, applied to biochemical equipment and methods, fixed on/in organic carriers, lyase, etc., can solve the problems of complex operation, high cost, microbial pollution, etc., and achieve process reaction specialization 1. High yield and good purity

Inactive Publication Date: 2012-07-04
广东乐尔康生物科技股份有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the need to use amino acids and protective reagents as raw materials, there are many reaction steps, long reaction time, complicated operation, especially optical resolution and environmental pollution; environmental pollution and high cost
[0014]3. Microbial fermentation method: construct microbial strains to express glutamine, glutamine synthetase and γ-glutamine transpeptidase, etc., use glutamic acid Fermentation and synthesis of raw materials such as ethylamine and ethylamine; it is necessary to obtain strains that can express relevant enzymes at a high level, and have a cell energy regeneration system; there are many studies, but no industrial production has yet been seen
[0015]4. Callus cell culture extraction method: use tea tree stem tip as explant culture to produce callus, improve the content of theanine in the tissue, and then use Extraction and purification by ion exchange method; the composition of the medium is complex, and nitrate and ethylamine need to be added in particular, and there is a problem of microbial contamination that is not easy to solve; the culture cycle of tea tree callus is long, the production process is slow, and the cost is high

Method used

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  • Production process for synthetizing L-theanine through biological immobilized enzyme catalysis

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Dissolve 16.9g of sodium glutamate, 20.3g of magnesium chloride hexahydrate, 61.1g of sodium hexametaphosphate, 8.54g of ethylamine hydrochloride and 55.1g of disodium adenosine triphosphate in purified water, and adjust the pH to 7.0 with 10% sodium hydroxide solution. Add 100KU solid-phase enzyme and dilute to 5000ml with pure water. (The substrate concentration is 20mmols / L, 20mmols / L, 20mmols / L, 20mmols / L and 20mmols / L) Heated in a water bath, stirred and reacted at 37°C, continuously added 10% sodium hydroxide solution to maintain the pH of the reaction solution =7.0. Samples were taken every 10 minutes to detect the content of sodium glutamate to determine the conversion rate of the reaction; when the conversion rate of L-theanine reached 97%, the reaction solution was separated from the solid-phase enzyme by filtration.

[0046] Slowly flow the filtrate through a strong acidic ion exchange resin (resin equivalent is 1 morl), and then elute with pure water. After...

Embodiment 2

[0050] Dissolve 42.25g of sodium glutamate, 50.75g of magnesium chloride hexahydrate, 152.75g of sodium hexametaphosphate, 21.35g of ethylamine hydrochloride and 137.75g of disodium adenosine triphosphate in purified water, and adjust the pH to 7.0 with 10% sodium hydroxide solution. Add 250KU solid-phase enzyme and dilute to 5000ml with pure water, so that the substrate concentration is 50mmols / L, 50mmols / L, 50mmols / L, 50mmols / L and 50mmols / L respectively. Heat in a water bath, keep stirring at 37°C, and continuously add 10% sodium hydroxide solution to maintain the pH of the reaction solution at 7.0. Samples were taken every 10 minutes to detect the content of sodium glutamate to determine the conversion rate of the reaction; when the conversion rate of L-theanine reached 97%, the reaction solution was separated from the solid-phase enzyme by filtration.

[0051] Slowly flow the filtrate through a strong acidic ion exchange resin (resin equivalent: 2.5mol), and then elute wi...

Embodiment 3

[0055] Dissolve 84.5g of sodium glutamate, 101.5g of magnesium chloride hexahydrate, 305.5g of sodium hexametaphosphate, 242.7g of ethylamine hydrochloride and 275.5g of disodium adenosine triphosphate in purified water, and adjust the pH to 7.0 with 10% sodium hydroxide solution. Add 500KU solid-phase enzyme and dilute to 5000ml with pure water, so that the substrate concentration is 0.1mols / L, 0.1mols / L, 0.1mols / L, 0.1mols / L and 0.1mols / L. Heat in a water bath, keep stirring at 37°C, and continuously add 10% sodium hydroxide solution to maintain the pH of the reaction solution at 7.0. Samples were taken every 10 minutes to detect the content of sodium glutamate to determine the conversion rate of the reaction; when the conversion rate of L-theanine reached 97%, the reaction solution was separated from the solid-phase enzyme by filtration.

[0056] Slowly flow the filtrate through a strong acidic ion exchange resin (resin equivalent: 5.0mol), and then elute with pure water un...

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Abstract

The invention relates to a production process for synthetizing L-theanine through biological immobilized enzyme catalysis, which belongs to the technical field of biosynthesis. The L-theanine is synthetized through the biological immobilized enzyme catalysis by taking sodium glutamate, magnesium chloride hexahydrate, sodium hexametaphosphate, ethylamine hydrochloride and triphosadenine to serve as raw materials. The production process for synthetizing the L-theanine through the biological immobilized enzyme catalysis comprises the following process steps: (1) performing catalytic synthesis on a substrate and an immobilized enzyme to generate the L-theanine, and then, centrifugally separating the biological immobilized enzyme from reaction liquid; (2) removing anions and cations in the reaction liquid through cation exchange resin to generate purified L-theanine; (3) decoloring an L-theanine aqueous solution through activated carbon, and performing vacuum film concentrating to obtain an L-theanine concentrated solution; and (4) adding 95 percent of alcohol to the L-theanine concentrated solution to separate out white L-theanine precipitation crystals; centrifugally separating; and performing vacuum drying to obtain white L-theanine powder. The production process for synthetizing the L-theanine through the biological immobilized enzyme catalysis has the advantages of specific process reaction, high yield, high purity, short period, low energy consumption and the like and is suitable for industrial production.

Description

technical field [0001] The invention relates to a production process for synthesizing L-theanine catalyzed by a biological solid-phase enzyme, belonging to the technical field of biosynthesis. Background technique [0002] Common name: L-theanine, English name: L-Theanine; [0003] Chinese chemical name: (S)-2-amino-4-(N-ethyl formyl)butanoic acid; [0004] English chemical name: (S)-2-amino-4-(N-ethylcarbamoyl)butanoic acid; [0005] CA substance code: CAS: 3081-61-6; [0006] Structural formula: [0007] Molecular formula: C 7 h 14 N 2 o 3 ; [0008] Molecular weight: 174.20; [0009] Properties: This product is white crystalline powder, odorless and slightly sweet. [0010] Theanine L-Theanine is a unique free amino acid in tea. Theanine is glutamic acid γ-ethylamide, which has a sweet taste. The content of theanine varies with the variety and part of the tea. Theanine accounts for 1-2% by weight in dry tea. Theanine is similar in chemical structure to gl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/04C12N15/60C12N15/70C12N9/88C12N11/02
Inventor 周兴华范永军周丙午
Owner 广东乐尔康生物科技股份有限公司
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