脂肪酶催化外消旋对羟基苯甘氨酸甲酯水解制备对映体纯D-对羟基苯甘氨酸的新方法  

Lipase-Catalyzed Hydrolysis of Racemic p-Hydroxyphenylglycine Methyl Ester as a New Way for Preparation of Enantiopure D-p-Hydroxyphenylglycine

作　　者：张媛媛[1] 宗敏华[1] 娄文勇[1] 林颖敏[1] 吴虹[1] 

机构地区：[1]华南理工大学生物工程系,广东广州510640

出　　处：《催化学报》2005年第2期106-110,共5页

基　　金：广东省自然科学基金资助项目 (0 2 0 83 9) .

摘　　要：基于在反应体系中添加碳酸氢铵可有效地促进酶促外消旋对羟基苯甘氨酸甲酯不对称水解这一有趣现象的发现 ,探讨了利用有机介质中酶促外消旋对羟基苯甘氨酸甲酯不对称水解制备对映体纯 D 对羟基苯甘氨酸及其衍生物的可能性 ,研究了不同来源的酶、摇床转速、水含量、对羟基苯甘氨酸甲酯浓度、反应介质及温度等因素对该反应的影响 .结果表明 ,在所研究的 11种酶中脂肪酶Novozym 4 35对该反应的催化活性和对映体选择性较高 ,叔丁醇为最合适的反应介质 ,最适水含量为0 4 % (V/V) ,对羟基苯甘氨酸甲酯的适宜浓度为 2 0mmol/L ,最适摇床转速和反应温度分别为 130r/min和 30℃ .在此优化条件下反应 2 6h后 ,底物转化率和产物ee分别为 39 8%和 95 2 % .On the basis of the interesting discovery that ammonium bicarbonate could effectively enhance the enzymatic asymmetric hydrolysis of racemic p-hydroxyphenylglycine methyl ester to D-p-hydroxyphenylglycine, the possibility of preparation of enantiopure D-p-hydroxyphenylglycine by this reaction has been explored for the first time. Effects of enzyme sources, shaking rate, water content, p-hydroxyphenylglycine methyl ester concentration, reaction media and reaction temperature on the reaction were examined systematically. Among the eleven enzymes assayed, Novozym 435 showed both the highest catalytic activity and the highest enantioselectivity. Tert-butanol was the most suitable medium for the reaction. The optimum water content, p-hydroxyphenylglycine methyl ester concentration, shaking rate and reaction temperature were 0.4% (V/V), 20 mmol/L, 130 r/min and 30 degrees C, respectively. Under the optimal conditions as above, a product enantiomeric excess (ee) of 95.2% was achieved with a substrate conversion of 39.8% after 26 h reaction time.

关 键 词：脂肪酶 Novozym 435 对羟基苯甘氨酸甲酯 不对称水解 D-对羟基苯甘氨酸 有机介质 

分 类 号：TQ463[化学工程—制药化工]