Preparation method of S-adenosylmethionine (SAMe) by enzyme-catalyzed method

A technology of adenosylmethionine and methionine, which is applied in the field of enzymatic synthesis and preparation of adenosylmethionine, can solve the problem of high speed, short reaction time, and high cost of SAMe synthase immobilization carrier problems, to achieve the effect of increased accumulation, fast SAMe conversion, and cost savings of carriers

Inactive Publication Date: 2016-12-07
BEIJING KAWIN TECH SHARE HLDG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] When utilizing SAMe synthetase to synthesize SAMe in the prior art, the problem that the cost of SAMe synthetase immobilized carrier is too high, the object of the present invention is to provide a kind of method that uses carrier-free immobilized SAMe synthetase enzymatically to synthesize SAMe, the The method is immobilized in the form of cross-linked enzyme aggregates, which not only has no immobilized carrier cost, but also requires a short reaction time, high speed, and high product yield. After the immobilized enzyme is used many times, the ability to convert SAMe is not reduced. Suitable for industrial production needs

Method used

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  • Preparation method of S-adenosylmethionine (SAMe) by enzyme-catalyzed method
  • Preparation method of S-adenosylmethionine (SAMe) by enzyme-catalyzed method
  • Preparation method of S-adenosylmethionine (SAMe) by enzyme-catalyzed method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1: Preparation of SAMe synthetase solution can be prepared by referring to the method of Chinese patent CN101979645B.

Embodiment 2

[0029] Example 2 Determination of Enzyme Activity

[0030] Composition of bioassay reaction solution: ATP 10mM, L-methionine 20mM, KCl 50mM, MgCl 2 50mM, 100mM Tris-HCl buffer, pH7.5. The reaction system is 100ml, the reaction temperature is 40°C, and the reaction is 30min. When finishing the reaction, the free enzyme uses 1.0ml concentrated hydrochloric acid to finish the reaction, and the immobilized enzyme uses the membrane separation enzyme and the reaction liquid to finish the reaction.

Embodiment 3

[0031] Example 3 Preparation of SAMe synthetase cross-linked enzyme aggregates

[0032]One volume of SAMe synthetase bacteriostasis solution (specific activity 90U / ml), was added with solid ammonium sulfate to make the saturation of ammonium sulfate reach 60%. Stir for 30 min, add glutaraldehyde reagent to make the final concentration reach 1.0%, stir and cross-link for 8.0 h. The above operations are carried out below 10°C (above 0°C, such as around 4°C). Centrifuge to obtain cross-linked enzyme aggregates and immobilized enzymes for later use.

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Abstract

The invention relates to a method for producing S-adenosylmethionine (or adenosyl methionine) by enzyme catalysis. SAMe synthases are prepared to form cross-linked enzyme aggregates, and the cross-linked enzyme aggregates form immobilized enzymes which are not soluble in water, therefore, the immobilized enzymes are prepared by the method to catalyze adenosine triphosphate (ATP) and methionine to synthesize adenosyl methionine. The preparation method of S-adenosylmethionine (SAMe) by the enzyme-catalyzed method disclosed by the invention is high in synthesis speed because the reaction can be completed by about 3 hours, high in synthetic quantity, stable in catalytic performance and not decreased in conversion capability after 60 times of reaction.

Description

technical field [0001] The invention relates to an enzymatic synthesis and preparation method of adenosylmethionine, in particular to the preparation of adenosylmethionine by catalytic synthesis with immobilized adenosylmethionine synthetase, and belongs to the field of biopharmaceuticals. Background technique [0002] China is a country with a large population, but also a "big hepatitis country". According to the national viral hepatitis epidemiological survey in 1992, about 120 million people in the country carry hepatitis B virus, and the infection rate of hepatitis C in the general population is 3.1%. There are currently 20 million chronic hepatitis B patients in my country, some of whom may transform into liver cirrhosis or even liver cancer. [0003] Adenosylmethionine (SAMe) is an extremely important intermediate metabolite in the human body. It can generate the key antioxidant and detoxification substance in the body-cysteine ​​through the transsulfurization pathwa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/40C12N11/18
CPCC12N11/18C12P19/40C12Y205/01006Y02P20/50
Inventor 付大雁曲智慧李岩侯建华
Owner BEIJING KAWIN TECH SHARE HLDG
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