细胞破碎方法对Bacillus sp. DL-2胞内蛋白酶拆分乙酸苏合香酯的影响

(R)-1-苯乙醇是手性药物合成的关键手性砌块, 在多种手性化合物的合成及医药、香料的工业生产中具有重要的作用。对筛选自西太平洋深海沉积物的Bacillus sp. DL-2进行破碎, 获取的胞内蛋白酶能有效拆分(±)-乙酸苏合香酯, 制备高光学纯的(R)-1-苯乙醇。为使拆分效果最优化, 本试验对比了表面活性剂、溶菌酶和超声法3种不同的破碎细胞方式, 并对表面活性剂和溶菌酶破碎法进行了单因素条件优化。最优方法及优化的反应条件为: 使用体积分数0.5%的表面活性剂曲拉通X-100在35℃和pH7.0条件下破碎细胞2h。经过优化后, (R)-1-苯乙醇的对映体过量值为96%, 转化率为23%, 为利用胞内蛋白酶生产手性化学品提供了参考。

The effects of different cell breaking methods on the asymmetrical hydrolysis of phenylethyl acetate using intracellular proteases of Bacillus sp.DL-2

(R)-1-phenylethanol is a key chiral building block in the synthesis of chiral drugs,and plays important roles in the synthesis of many chiral compounds and in the industrial production of medicines and spices.The strain Bacillus sp.DL-2 screened from the deep-sea sediments of the western Pacific Ocean was broke,and the obtained intracellular proteases can asymmetrically hydrolyze(±)-1-phenylethyl acetate to obtain(R)-1-phenylethanol.Three different ways,namely,surfactant,lysozyme and ultrasonic,were utilized to crush Bacillus sp.DL-2.After the optimizations of the surfactant and lysozyme crushing methods,the optimal working conditions were found to be 0.5%TritonX-100,35℃and pH 7.0 for 2 h.After process optimization,(R)-1-phenylethanol was obtained with an enantiomeric excess of 96%and a conversion of 23%,which provide reference for using intracellular proteases to produce chiral chemicals.