Method for preparing S-ademetionine and D-methionine

A technology for adenosylmethionine and methionine, which is applied in the field of preparing S-adenosylmethionine and D-methionine, can solve the problem of high cost of chiral resolution, achieve low production cost, eliminate waste problems, and realize the effect of recycling

Inactive Publication Date: 2015-09-23
NINGBO INST OF TECH ZHEJIANG UNIV ZHEJIANG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although the cost of chemically synthesizing D/L-methionine is low, the cost of chiral resolution is relatively high. Therefore, the price of D-methionine is not...

Method used

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  • Method for preparing S-ademetionine and D-methionine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Saccharomyces cerevisiae (Saccharomyces cerevisiae), which was cryopreserved at 4°C and producing SAM synthase, was streak-inoculated into a solid activated medium, cultured at 30°C overnight, transferred to a liquid seed medium, and then incubated at 30°C with a rotating speed. After culturing overnight on a shaker at 250 rpm, fresh seeds were inoculated into a fermenter at an inoculum of 6% by volume, and a 2L liquid fermentation medium was placed in the fermenter, and cultured at 28°C. The rotating speed of the stirring paddle of the fermentation tank is 150rpm, and the ventilation rate is 250m 3 / h, the pH value of the fermentation broth was kept at 5.0.

[0037] Wait for the strain to grow to mid-logarithmic growth OD 600 After = 25, add 4g D / L-methionine to the fermentation broth one by one, adding 6 times in total, stop after the L-methionine in the fermentation broth is completely consumed at the end of the fermentation, and centrifuge the fermentation broth at...

Embodiment 2

[0039] Take the bacterial cells collected in Example 1, wash the bacterial cells twice with normal saline, resuspend the bacterial cells to 200 mL with a mass fraction of 3% sulfuric acid solution, and incubate at 30 °C for 10 h to fully lyse the bacteria. Somatic cells to release intracellular S-adenosylmethionine, and then centrifuged at 4000 rpm for 15 min, discarded cell debris pellets, collected the supernatant after breaking cells, and adjusted the pH value to 5.0 ± 0.5.

[0040] Take HZD-2 resin, rinse it with tap water for 5 times, remove mechanical impurities, and soak the resin with NaOH solution with a mass fraction of 2 times the resin volume and a mass fraction of 10%. After 2 hours, wash with deionized water until neutral, soak the resin with 10% hydrochloric acid for 2 hours to transform the resin, wash with deionized water until neutral, and then pack the chromatographic column; The supernatant (pH value: 5.0±0.5) collected by centrifugation was pumped into the...

Embodiment 3

[0042] Take D018 resin, rinse it with tap water for 5 times to remove mechanical impurities, soak the resin with 10% sodium hydroxide in a mass fraction of 2 times the volume of the resin, and after 2 hours, wash it with deionized water until it becomes neutral, and then use a mass fraction of Soak in 10% hydrochloric acid for about 2 hours, and wash with deionized water until neutral. After repeating the above steps once, the resin was packed into the chromatographic column, rinsed with 10% hydrochloric acid with a mass fraction of 4 times the resin volume, and washed with deionized water until the pH value of the effluent was neutral; / L of ammonia water, soak the resin with the same concentration of ammonia water to convert the resin into NH 4 + type, wash the column with deionized water until the pH of the effluent is neutral. Then, the fermentation broth supernatant collected in Example 1 was pumped into the treated D018 resin chromatographic column at a flow rate of 1 ...

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Abstract

The invention discloses a method for preparing S-ademetionine and D-methionine. The method comprises the following steps: fermenting D/L-methionine which serves as a raw material to obtain the S-ademetionine; separating the S-ademetionine from bacteria cells; separating the D-methionine from fermentation liquor. According to the method, the low-price D/L-methionine serves as the raw material, strains containing S-adenosylmethionine synthetase are adopted for fermentation, the L-methionine reacts with ATP in the bacteria cells to generate the S-ademetionine, the S-ademetionine is separated from the D-methionine through a cell membrane, and the S-ademetionine and the D-methionine are separated in separation manners such as a centrifugal separation manner and an ion exchange manner respectively; according to the method, the S-ademetionine and the D-methionine can be synchronously produced, and the D-methionine can be recycled.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for preparing S-adenosylmethionine and D-methionine. Background technique [0002] S-adenosyl-L-methionine (SAM) is an important physiologically active substance. As a methyl donor and a precursor of physiological sulfhydryl compounds, it is involved in the transfer of methyl groups and the Various biochemical reactions related to sulfhydryl transfer play an indispensable role in maintaining the normal mobile phase of the liver cell membrane and the detoxification function of the liver. [0003] Studies have found that SAM has a positive therapeutic effect on various diseases such as intrahepatic cholestasis, cirrhosis, depression and osteoarthritis caused by liver diseases such as viral hepatitis, alcoholic liver, fatty liver, etc. It has been used in Italy, Germany, Spain, etc. The state is approved as a prescription drug to enter clinical use. [0004] In my country, SA...

Claims

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Application Information

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IPC IPC(8): C12P41/00C12P19/40C12P13/12C12R1/865
Inventor 胡升金志华梅乐和吴志革杨郁尚龙安
Owner NINGBO INST OF TECH ZHEJIANG UNIV ZHEJIANG
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