摘要
对重组毕赤酵母胞内S-腺苷-L-甲硫氨酸（SAM）含量以及SAM合成酶活性的高效液相色谱（HPLC）测定方法进行了优化。通过调整流动相浓度和洗脱程序,建立了保留时间适中、分离效果好的SAM定量分析方法。结果表明,SAM浓度在0.05~0.5 g.L-1时,其峰面积与浓度线性关系良好,相关系数为0.99996,平均回收率为99.3%~100.5%,相对标准偏差为0.72%~0.98%。该方法对胞内以及酶活测定液中的SAM能够进行有效分离,精密度高、稳定性好,能高效、准确地分析发酵过程菌体中SAM含量以及SAM合成酶活性。
The method for determination of S-adenosyl-L-methionine（SAM） content in cell extract of recombinant Pichia pastoris,as well as SAM synthetase activity of the enzymatic reaction mixture,by HPLC was studied.The composition of mobile phase and the gradient elution program were optimized to achieve a high resolution with a moderate retention time of SAM.As a result,a good linearity was observed for the calibration graph（R2=0.99996） in the range of 0.05 ~ 0.5 g·L-1 for SAM.The recoveries of SAM were 99.3%~100.5% and the relative standard deviation were 0.72%~0.98%.SAM in the cell extract sample or enzymatic reaction mixture could be separated effectively and determined accurately.
出处
《化学与生物工程》
CAS
2011年第6期83-86,91,共5页
Chemistry & Bioengineering
基金
国家自然科学基金资助项目(20976050)
