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Method for preparing uridine diphosphate glucose by means of immobilized enzyme method

A technology of uridine diphosphate, immobilized enzyme, applied in biochemical equipment and methods, immobilized on or in inorganic carriers, immobilized on/in organic carriers, etc., can solve problems affecting practical application value, Inability to effectively recycle, rapid decline in enzyme activity, etc., to achieve the effects of stable product quality, avoidance of impurities, and easy reaction

Pending Publication Date: 2020-02-28
美亚药业海安有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method uses free enzymes to produce uridine diphosphate glucose, and the enzyme activity decreases rapidly, which cannot be effectively recycled, which will inevitably lead to high production costs and affect the actual application value.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] A method for preparing uridine diphosphate glucose by an immobilized enzyme method, specifically comprising the following steps:

[0019] (1) Preparation of UDPG production enzyme: UDPG production enzyme is immobilized on an immobilized carrier, using uridine diphosphate glucose pyrophosphorylase (EC 2.7.7.9, UGP), sucrose phosphorylase (EC 2.4.1.7, SPA) Two enzymatic reactions can be synthesized to prepare immobilized UDPG production enzyme;

[0020] (2) Immobilized UDPG production enzyme: use immobilized UDPG production enzyme to catalyze the preparation of uridine diphosphate glucose reaction liquid, filter and collect the carrier to obtain immobilized UDPG production enzyme;

[0021] (3) Separation of products: the above-mentioned immobilized UDPG production enzyme is directly separated in the reaction tank, and the separated reaction solution is recovered through a filter bag to recover the immobilized UDPG production enzyme, and the permeate is separated by chroma...

Embodiment 2

[0026] A method for preparing uridine diphosphate glucose by an immobilized enzyme method, specifically comprising the following steps:

[0027] (1) Preparation of UDPG production enzyme: UDPG production enzyme is immobilized on an immobilized carrier, using uridine diphosphate glucose pyrophosphorylase (EC 2.7.7.9, UGP), sucrose phosphorylase (EC 2.4.1.7, SPA) Two enzymatic reactions can be synthesized to prepare immobilized UDPG production enzyme;

[0028] (2) Immobilized UDPG production enzyme: use immobilized UDPG production enzyme to catalyze the preparation of uridine diphosphate glucose reaction liquid, filter and collect the carrier to obtain immobilized UDPG production enzyme;

[0029] (3) Separation of products: the above-mentioned immobilized UDPG production enzyme is directly separated in the reaction tank, and the separated reaction solution is recovered through a filter bag to recover the immobilized UDPG production enzyme, and the permeate is separated by chroma...

Embodiment 3

[0034] A method for preparing uridine diphosphate glucose by an immobilized enzyme method, specifically comprising the following steps:

[0035] (1) Preparation of UDPG production enzyme: UDPG production enzyme is immobilized on an immobilized carrier, using uridine diphosphate glucose pyrophosphorylase (EC 2.7.7.9, UGP), sucrose phosphorylase (EC 2.4.1.7, SPA) Two enzymatic reactions can be synthesized to prepare immobilized UDPG production enzyme;

[0036](2) Immobilized UDPG production enzyme: use immobilized UDPG production enzyme to catalyze the preparation of uridine diphosphate glucose reaction liquid, filter and collect the carrier to obtain immobilized UDPG production enzyme;

[0037] (3) Separation of products: the above-mentioned immobilized UDPG production enzyme is directly separated in the reaction tank, and the separated reaction solution is recovered through a filter bag to recover the immobilized UDPG production enzyme, and the permeate is separated by chromat...

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PUM

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Abstract

The invention relates to the technical field of biology, in particular to a method for preparing uridine diphosphate glucose by means of an immobilized enzyme method. The method specifically comprisesthe steps of: (1) preparing UDPG (uridine diphosphate glucose) production enzymes; (2) immobilizing the UDPG production enzymes; and (3) separating a product. According to the invention, two UDPG production enzymes UGP and SPA are adopted, and the UDPG can be synthesized by just two-step enzymatic reaction; compared with a complex process for producing UDPG by chemical and fermentation methods, the method here has a simpler reaction process, with the reaction easier to control, and the product quality being more stable; the UDPG is prepared by means of an immobilized enzyme catalysis method,and the immobilized enzymes can be continuously and repeatedly used many times, so that the production cost is greatly reduced; in addition, a large amount of pigments, other types of nucleotides andother impurities introduced by the use of yeast are avoided, so that purification is easier; the method is suitable for large-scale production of UDPG.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for preparing uridine-glucose diphosphate by an immobilized enzyme method. Background technique [0002] The synthesis of UDPG mainly includes chemical synthesis, fermentation and enzymatic conversion. Chemical synthesis requires the protection and deprotection of active groups on sugar groups, expensive catalysts, harsh reaction conditions, large amounts of organic solvents, serious environmental pollution, long reaction cycles, and low yields. Fermentation also has the disadvantage of low yield. Because the enzyme has strong stereoselectivity and specificity to the substrate, the enzymatic synthesis of UDPG becomes a simple and easy way. Moreover, the rapid development of gene recombination technology in recent years has enabled large-scale production of various enzymes at a lower cost, making the advantages of enzyme synthesis more significant. [0003] The enzymatic s...

Claims

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Application Information

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IPC IPC(8): C12P19/30C12N11/18C12N11/14C12N11/10
CPCC12P19/305C12N11/18C12N11/14C12N11/10C12N9/1241C12N9/1051C12Y207/07009C12Y204/01007
Inventor 邓壮梅王宏傅得响张燕
Owner 美亚药业海安有限公司
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