响应面法优化丁酸缩水甘油酯的酶法拆分工艺

Optimization of Enzymatic Resolution Technique of Glycidyl Butyrate via Response Surface Methodology

摘要:

在已有的酶法拆分丁酸缩水甘油酯单因素试验最适条件的基础上, 采用Plackett-Burrman设计在影响酶法拆分的6个因素中, 有效筛选出主效因素: 底物浓度、酶用量和温度。在此基础上, 再利用响应面分析法(RSM)对以上三个显著因子的最佳水平范围进行研究, 通过对二次多项回归方程求解得知, 酶法拆分最适条件为: 底物浓度0.499 mol/L、酶用量30.23 mg/(g底物)和温度29.68oC; 并结合单因素最适条件: pH 7.6; 时间4 h; 转速150 r/min进行酶促拆分实验, 得到R-酯的对映体过量值为93.28%。对比优化之前的单因素试验最适条件的结果, 最大对映体过量值84.65%, 有了显著的提高, 证明RSM法优化酶法拆分丁酸缩水甘油酯工艺是可行的。

Abstract:

Our previous work has indicated that enzymatic resolution of glycidyl butyrate are strongly affected by many factors, including concentration of substrate, amount of lipase, the temperature, pH, shaking speed and reaction time. In this study, Plackett-Burman design was undertaken to evaluate the effects of the six factors. By regression analysis, concentration of substrate, amount of lipase and the temperature were found to be important for enzymatic resolution of glycidyl butyrate. In the second phase of the optimization process, a response surfacemethodology(RSM) was used to optimize the above critical factors, and to find out the optimal concentration levels and the relationships between these factors. By solving the quadratic regression model equation using appropriate statistic methods, the optimal parameter of the variables were determined as: 0.499 mol/L glycidyl butyrate, 30.23 mg/g lipase and 29.68oC. In the optimum condition, the value of enantiomeric excess(ee%) was 93.28%. Compared to 84.65% which was the maximum ee% under the non-optimized condition, this study has a significant advancement. The experimental data under various conditions have validated the theoretical values.