响应面法优化丁酸缩水甘油酯的酶法拆分工艺-微生物学报.PDF

生 物 工 程 学 报 Chin J Biotech 2008, June 25; 24(6): 1062-1067 journals.im.ac.cn Chinese Journal of Biotechnology ISSN 1000-3061 cjb@im.ac.cn © 2008 Institute of Microbiology, CAS CSM, All rights reserved 研究报告 响应面法优化丁酸缩水甘油酯的酶法拆分工艺 钱俊青, 周文武, 匡春兰 浙江工业大学药学院, 杭州 310014 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。 关键词: 丁酸缩水甘油酯, 脂肪酶, 拆分, Plackett-Burrman 设计, 响应面分析法, 优化 Optimization of Enzymatic Resolution Technique of Glycidyl Butyrate via Response Surface Methodology Junqin Qian, Wenwu Zhou, and Chunlan Kuang College of Pharmaceut Science, Zhejiang University of Technology, Hangzhou 310014, China 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