Method for synthesizing S-adenosylmethionine through coupling of escherichia coli and saccharomyces cerevisiae

The technology of adenosylmethionine and Escherichia coli is applied in the field of bioengineering, which can solve the problems of difficulty in separation and purification and high cost, and achieve the effects of simplifying the separation and purification process, improving the yield and improving the utilization rate.

Inactive Publication Date: 2014-04-30
JIMEI UNIV
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  • Claims
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Problems solved by technology

[0005] The technical problem to be solved by the present invention is to provide a method for the above-mentioned deficiencies in the prior art, and to provide a preparation method for synthesizing S-adenosylmethionine by coupling E. There are problems such as high cost and difficulty in separation and purification

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  • Method for synthesizing S-adenosylmethionine through coupling of escherichia coli and saccharomyces cerevisiae
  • Method for synthesizing S-adenosylmethionine through coupling of escherichia coli and saccharomyces cerevisiae
  • Method for synthesizing S-adenosylmethionine through coupling of escherichia coli and saccharomyces cerevisiae

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Embodiment Construction

[0027] The present invention will be described in further detail below in conjunction with specific examples.

[0028] 1. Cultivation of Escherichia coli engineering strain BL21 / pET-28a-SAMS (constructed in our laboratory):

[0029] 1. Medium: LB medium (yeast powder 5 g / L, tryptone 10 g / L, NaCl 10 g / L, pH 7.0), solid LB medium contains 2% agar. Add 50 µg / mL kanamycin as needed.

[0030]2. Culture method: Pick a single colony of Escherichia coli engineering strain BL21 / pET-28a-SAMS from the solid LB medium containing 50 μg / mL kanamycin and inoculate it into a 30 mL medium containing 50 μg / mL kanamycin Cultivate in a 250 mL Erlenmeyer flask with mymycin LB (LK) medium, and cultivate for about 12 h at a shaker speed of 180 r / min and a temperature of 37 °C. The culture solution was inoculated into a 250 mL Erlenmeyer flask containing 100 mL of LK medium at an inoculation amount of 1% (v / v) for cultivation at a temperature of 37°C and a shaker speed of 180 r / min. When the bacter...

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Abstract

The invention discloses a method for synthesizing S-adenosylmethionine (SAM) through coupling of escherichia coli and saccharomyces cerevisiae. The method comprises basic processes of firstly, respectively cultivating an escherichia coli engineered strain BL21 / pET-28a-SAMS containing S-adenosylmethionine synthetase gene (SAMS) and saccharomyces cerevisiae at proper conditions, then mixing according to a certain proportion, coupling so as to generate SAM; secondly, allowing the synthesized SAM to secrete to outer side of the cell by taking an organic solvent as a permeability agent; and finally, analyzing and detecting the SAM by a high performance liquid chromatography technology. The invention provides a novel method for producing SAM at low cost, and the method is simple and convenient to operate, and can catalyze ATP and L-Met by the whole cell so as to synthesize SAM.

Description

technical field [0001] The invention belongs to the field of bioengineering, and in particular relates to a preparation method for synthesizing S-adenosylmethionine through the coupling of Escherichia coli and Saccharomyces cerevisiae. Background technique [0002] S-adenosylmethionine (S-Adenosyl-methionine, SAM) is an important intermediate metabolite widely present in animals, plants and microorganisms. In the human body, SAM is an important physiologically active substance that participates in various biochemical reactions. SAM has attracted widespread attention due to its important physiological functions. After years of clinical application research, it has been shown that SAM is the substance with the best effect, the most functions and the widest application range, which can be used not only as a disease treatment drug, but also as a food nutrition enhancer. It can be used to treat diseases such as liver disease, depression and arthritis, and is also an advanced he...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P39/00C12P19/40C12R1/19C12R1/865
Inventor 杜翠红魏晓楠
Owner JIMEI UNIV
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