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Novel aden osyl methionine in vitro enzymatic transformation method

A technology of adenosylmethionine and methionine, which is applied in the field of in vitro enzymatic conversion of adenosylmethionine, can solve the problems of manpower and material resources, not suitable for large-scale industrial production, and ATP instability

Active Publication Date: 2008-01-23
上海国佳生化工程技术研究中心有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the problem with in vitro enzymatic conversion is that the cost of commercially purchased ATP is extremely high, which is not suitable for large-scale industrial production; and ATP is very unstable and difficult to store
In addition, the traditional in vitro enzymatic conversion method needs to add ATP in batches, which not only needs to prevent its inactivation when adding, but also consumes manpower and material resources. Therefore, it is urgent to develop a S-adenosine that can reduce costs and is easy to operate Method for producing methionine

Method used

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  • Novel aden osyl methionine in vitro enzymatic transformation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0091] ATP production by free Saccharomyces cerevisiae and SAM synthesis reaction involving free SAM synthetase

[0092] 1 material

[0093] 1.1 ATP producing strain: the strain used in this example is Saccharomyces cerevisiae CICC 1012.

[0094] 1.2 SAM synthetase producing strain: The SAM synthetase producing strain used in this example has a strain preservation number of CGMCC, NO.0648, provided by the Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, and its construction process can be found in CN1357630A.

[0095] 1.3 Lysis buffer:

[0096] Phosphate buffer 50mM / L, glycerin 5%, EDTA 1mM / L, PMSF 0.5mM / L

[0097] 1.4 Bioreactor: FUS-15L fermenter, produced by Shanghai Guoqiang Biochemical Engineering Equipment Co., Ltd.

[0098] 1.5 Medium: YPD medium: yeast extract 10g / L, peptone 20g / L, glucose 20g / L.

[0099] 1.6 Mixed reaction solution

[0100] L-methionine 10mM / L, glutathione 16mM / L, dipotassium hy...

Embodiment 2

[0112] ATP production by immobilized Saccharomyces cerevisiae and SAM synthesis reaction involving immobilized SAM synthetase

[0113] 1. Materials

[0114] 1.1 ATP producing strain: the strain used in this example is Saccharomyces cerevisiae CICC 1202.

[0115] 1.2 The production strain of SAM synthetase, yeast lysate buffer, bioreactor or shake flask, and YPD medium required for yeast growth are all the same as in Example 1.

[0116] 1.3 The reaction solution for coupling immobilized yeast with immobilized SAM synthetase contains:

[0117] L-methionine 10mM / L, glutathione 16mM / L, potassium chloride 200mM / L, dipotassium hydrogen phosphate 200mM / L, Ado 40mM / L, glucose 150mM / L, magnesium sulfate 20mM / L, Tris - HCl buffer 10 mM / L; reaction pH 7.0.

[0118] 2 methods

[0119] 2.1 The preparation of SAM synthetase is the same as in Example 1.

[0120] 2.2 Immobilization of SAM synthetase

[0121] Dilute the initially obtained crude enzyme solution with a pH 6.8 phosphate buf...

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Abstract

The invention provides the new method of composing the SAM by means of the enzyme, (1) using the ATP to producing the bacterium individual plant which can fordizethe ATP; (2) transforming the SAM composing enzyme to produce SAM; the said two actions are connected. The method improves the efficiency of the ATP producing bacterium individual plant producing the ATP greatly and depressing the cost of the ectoenzym out of the cell producing the SAM. So it is the same with the mass production of the SAM.

Description

technical field [0001] The invention relates to the field of biochemical industry, more specifically, the invention relates to an in vitro enzymatic conversion method of adenosylmethionine (SAM). Background technique [0002] As we all know, S-adenosyl-L-methionine (S-adenosyl-L-methionine, SAM) is an important metabolic intermediate widely present in animals, plants and microorganisms, and is also an important physiologically active substance in the human body. More than 40 kinds of biochemical reactions are important multi-curative drugs. [0003] At present, the preparation methods of S-adenosylmethionine mainly include chemical synthesis, biotransformation and in vitro enzymatic conversion. In vitro enzymatic conversion is to use S-adenosylmethionine synthetase to catalyze the substrate L-methionine (L-Met) and adenosine triphosphate (ATP) to generate S-adenosylmethionine in vitro. Compared with chemical synthesis and biotransformation, in vitro enzymatic conversion ha...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P19/32
Inventor 庄英萍武鑫储炬张嗣良王永红胡晓清袁中一
Owner 上海国佳生化工程技术研究中心有限公司
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