在水/有机溶剂双相体系中,利用重组大肠杆菌DM206 [pET28a-trpBA/BL21 (DE3)]色氨酸合成酶催化L-丝氨酸和吲哚合成了L-色氨酸.考察了有机溶剂、有机相乙酸乙酯体积分数、反应温度、水相pH、底物L-丝氨酸与吲哚摩尔比、底物L-丝氨酸浓度、表面活性剂和酶添加量对色氨酸合成酶酶活的影响.结果表明,酶法最佳转化条件为:有机相溶剂为乙酸乙酯,乙酸乙酯的体积分数为2.5%,反应温度为35℃,水相pH值为8,底物L-丝氨酸与吲哚的摩尔比为1∶1.2,底物L-丝氨酸浓度为100mmol/L,吐温80的质量分数为0.4%,反应介质中酶的添加量为20g/L.在此条件下酶促反应2.5 h,L-丝氨酸转化率可达95%.
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