利用甲醇传感器及高效液相色谱检测毕赤酵母摇瓶发酵过程的甲醇浓度及S-腺苷蛋氨酸(SAM)浓度,发现L-蛋氨酸浓度及甲醇浓度对毕赤酵母细胞生长及合成S-腺苷蛋氨酸具有影响,据此对摇瓶发酵过程的L-蛋氨酸浓度及甲醇浓度进行优化。优化结果表明:当L-蛋氨酸浓度为7.5 g/L时,最适于SAM积累,产量达到0.83 g/L;进而利用甲醇传感器对发酵过程的甲醇浓度进行检测及控制,考察不同甲醇浓度对SAM产量的影响,毕赤酵母产SAM的最佳甲醇浓度为15 g/L,在此浓度下SAM的产量达到1.41 g/L,比对照实验增加了21%。
It was found that the concentrations of L-methionine and methanol possessed effects on the cell growth of Pichia pastoris and the synthesis of S-adenosyl-L-methionine(SAM) adopting methanol sensor and HPLC to test and determine both concentrations in the shake-flask fermentation process;in accordance with this discovery an optimization of both concentrations of L-methionine and methanol were carried out in shake-flask fermentation process.The optimized results showed that,when the L-methionine concentration was at 7.5 g/L,it was the most suitable for the accumulation of SAM,and the yield reached as much as 0.83 g/L.Methanol sensor was further used to test and determine,and control the concentration of methanol to observe the effect of methanol concentration on SAM production.The optimal concentration of methanol for P.pastoris to produce SAM at was at 15 g/L,under this concentration it reached as much as 1.41 g/L,increased by 21% as compared with the control experiments.