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Method for preparing triphosadenine by immobilized enzyme method

A technology of adenosine triphosphate and dry products of adenosine triphosphate, applied to biochemical equipment and methods, fixed on/in organic carriers, enzymes, etc., can solve the problem of high cost, reduce raw material costs, and be easy to control the reaction and purify Effect

Pending Publication Date: 2020-02-11
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

However, this method uses AMP as the substrate, and the cost of the substrate is much higher than that of adenosine (Ar).

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] A method for preparing adenosine triphosphate by an immobilized enzyme method, specifically comprising the following steps:

[0018] (1) Preparation of ATP-producing enzymes: ATP-producing enzymes were immobilized on an immobilized carrier, and adenosine kinase (EC2.7.1.20, Ark), adenylate kinase (EC 2.7.4.3, Adk) and polyphosphate- Adenylate phosphotransferase (EC2.7.4.-, Pap) can be synthesized by three enzymatic reactions, and ATP is prepared by immobilized ATP production enzyme;

[0019] (2) Immobilized ATP-producing enzyme: using the immobilized ATP-producing enzyme, adenosine is catalyzed to prepare an adenosine triphosphate reaction solution, and the carrier is collected by filtration to obtain the immobilized ATP-producing enzyme;

[0020] (3) Separation of products: directly separate the above-mentioned immobilized ATP production enzyme in the reaction tank, recover the immobilized ATP production enzyme from the separated reaction solution through a filter bag,...

Embodiment 2

[0025] A method for preparing adenosine triphosphate by an immobilized enzyme method, specifically comprising the following steps:

[0026] (1) Preparation of ATP-producing enzymes: ATP-producing enzymes were immobilized on an immobilized carrier, and adenosine kinase (EC2.7.1.20, Ark), adenylate kinase (EC 2.7.4.3, Adk) and polyphosphate- Adenylate phosphotransferase (EC2.7.4.-, Pap) can be synthesized by three enzymatic reactions, and ATP is prepared by immobilized ATP production enzyme;

[0027] (2) Immobilized ATP-producing enzyme: using the immobilized ATP-producing enzyme, adenosine is catalyzed to prepare an adenosine triphosphate reaction solution, and the carrier is collected by filtration to obtain the immobilized ATP-producing enzyme;

[0028] (3) Separation of products: directly separate the above-mentioned immobilized ATP production enzyme in the reaction tank, recover the immobilized ATP production enzyme from the separated reaction solution through a filter bag,...

Embodiment 3

[0033] A method for preparing adenosine triphosphate by an immobilized enzyme method, specifically comprising the following steps:

[0034] (1) Preparation of ATP-producing enzymes: ATP-producing enzymes were immobilized on an immobilized carrier, and adenosine kinase (EC2.7.1.20, Ark), adenylate kinase (EC 2.7.4.3, Adk) and polyphosphate- Adenylate phosphotransferase (EC2.7.4.-, Pap) can be synthesized by three enzymatic reactions, and ATP is prepared by immobilized ATP production enzyme;

[0035] (2) Immobilized ATP-producing enzyme: using the immobilized ATP-producing enzyme, adenosine is catalyzed to prepare an adenosine triphosphate reaction solution, and the carrier is collected by filtration to obtain the immobilized ATP-producing enzyme;

[0036](3) Separation of products: directly separate the above-mentioned immobilized ATP production enzyme in the reaction tank, recover the immobilized ATP production enzyme from the separated reaction solution through a filter bag, ...

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PUM

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Abstract

The invention relates to the field of a biological technology, in particular to a method for preparing triphosadenine by an immobilized enzyme method. The method particularly comprises the following steps: (1) preparing ATP production enzyme; (2) immobilizing the ATP production enzyme; and (3) separating products. The ATP can be synthesized by adopting three kinds of ATP production enzyme such asadenosine kinase (EC 2.7.1.20, Ark), adenylate kinase (EC 2.7.4.3, Adk) and polyphosphoric acid-adenylate phosphotransferase (EC 2.7.4.-, Pap) and only through two-step enzymatic reaction, the reaction process is simple, the reaction is easy to control and the product quality is more stable; the ATP is prepared by an immobilized enzyme catalysis method, the immobilized enzyme can be used continuously and repeatedly for many times, and the production cost is greatly reduced; meanwhile, introduction of a large amount of pigment and other types of nucleotide impurities caused by the use of yeastis avoided and purification is facilitated; and the method is suitable for producing the ATP on a large scale.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for preparing adenosine triphosphate by an immobilized enzyme method. Background technique [0002] Utilizing the glycolysis pathway of yeast and using adenosine as a substrate to phosphorylate and synthesize ATP at the substrate level is the most commonly used method, and it is also the method commonly used in the industrial production of adenosine triphosphate (ATP). However, the reaction process of synthesizing ATP catalyzed by yeast cell enzymes is complex, and there are many enzymes involved in the catalytic reaction, the reaction process is difficult to control, and the quality of products varies greatly between batches. At the same time, the quality of yeast enzymes often varies greatly due to different suppliers, different batches, and even different seasons. The quality of yeast enzymes is unstable, the enzyme activity decreases rapidly, and the service life is sho...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/32C12N11/18C12N11/10C12N9/12
CPCC12P19/32C12N11/18C12N11/10C12N9/1205C12N9/1229C12Y207/0102C12Y207/04003
Inventor 邓壮梅徐超男王宏傅得响张燕
Owner 美亚药业海安有限公司
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