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
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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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