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Enzyme preparation for preparing citicoline, and method for preparing citicoline through enzyme catalysis

A technology for citicoline and enzyme preparation, which is applied in the field of preparation of citicoline and enzyme catalyzed preparation of citicoline, can solve the problem of toxic reagents and low synthesis yield, conversion rate and reaction rate The problems of low cost and high raw material cost can achieve the effect of fast reaction speed, high conversion rate and low raw material cost.

Active Publication Date: 2021-03-12
TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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Problems solved by technology

[0006] In view of the problems of toxic reagents and low synthesis yield in the existing chemical method, and the problems of high raw material cost, low conversion rate and reaction rate in the multi-enzyme catalytic method, this patent application is hereby proposed

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  • Enzyme preparation for preparing citicoline, and method for preparing citicoline through enzyme catalysis
  • Enzyme preparation for preparing citicoline, and method for preparing citicoline through enzyme catalysis
  • Enzyme preparation for preparing citicoline, and method for preparing citicoline through enzyme catalysis

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Embodiment Construction

[0046] Below in conjunction with embodiment, the present invention is further described, and following embodiment is descriptive, not limiting, can not limit protection scope of the present invention with following embodiment.

[0047] The raw materials used in the present invention, if no special instructions, are conventional commercially available products, the methods used in the present invention, if no special instructions, are conventional methods in this area, and the quality of each material used in the present invention is routine use quality.

[0048] An enzyme preparation for preparing citicoline, the preparation steps are as follows:

[0049] First, construct the engineering strain Escherichia coli CKI-CCT to express choline kinase and phosphorylcholine cytidine transferase derived from Fusobacterium nucleatum. Choline kinase can catalyze the formation of phosphorylcholine from choline chloride, and the transfer of phosphorylcholine cytidine The enzyme can cataly...

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Abstract

The invention relates to an enzyme preparation for preparing citicoline. The preparation method comprises the following steps: constructing engineering bacteria Escherichia coli CKI-CCT to express choline kinase and phosphorylcholine cytidyltransferase derived from fusobacterium nucleatum; constructing engineering bacteria Escherichia coli PPK-UDK to express polyphosphate kinase derived from rhodobacter sphaeroides and cytidine kinase derived from thermophilic thermophile; constructing engineering bacteria Escherichia coli CMK-NDK to express cytidylate kinase and nucleoside diphosphate kinasederived from escherichia coli; and crushing enzyme-producing cells obtained by fermentation, and centrifugally collecting supernatant to obtain a crude enzyme solution. According to the invention, itis reported that phosphorylcholine cytidine transferase with the fusobacterium nucleatum source can be used for synthesizing citicoline for the first time, and the heterologous soluble expression quantity and the enzyme activity of the phosphorylcholine cytidine transferase with the fusobacterium nucleatum source are higher.

Description

technical field [0001] The invention belongs to the technical field of compound biology, in particular to an enzyme preparation for preparing citicoline and a method for preparing citicoline by enzymatic catalysis. Background technique [0002] Citicoline is an important precursor for lecithin synthesis in organisms. Citicoline was first successfully developed by Japan's Takeda Pharmaceutical Co., Ltd., and its trade name is "Nicholin" (Nicolin). It has been included in the Chinese Pharmacopoeia as a drug for improving cell metabolism. Citicoline is the most clinically used Nerve activator, can promote the biosynthesis of lecithin, restore nerve tissue function, improve brain metabolism and circulation, can be used for recovery of consciousness disturbance caused by acute craniocerebral trauma and brain surgery, and can be used for Parkinson's syndrome and senile dementia adjuvant treatment of lecithin. Citicoline is the precursor of lecithin biosynthesis. When the brain fu...

Claims

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Application Information

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IPC IPC(8): C12N9/12C12N15/70C12P19/30C12R1/19
CPCC12N9/1205C12N9/1229C12N9/1241C12N15/70C12P19/305C12Y207/01032C12Y207/07015C12Y207/04001C12Y207/04025C12Y207/04006
Inventor 范晓光胡晨龙宋雪静安俊侠谢希贤徐庆阳
Owner TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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