Detry Jean, Rosenbaum Thorsten, Lütz Stephan, Hahn Doris, Jaeger Karl-Erich, Müller Michael, Eggert Thorsten
Institut für Biotechnologie 2, Forschungszentrum Jülich, D-52425 Jülich, Germany.
Appl Microbiol Biotechnol. 2006 Oct;72(6):1107-16. doi: 10.1007/s00253-006-0391-9. Epub 2006 Apr 4.
Two extracellular lipases from Bacillus subtilis, B. subtilis lipase A and lipase B, have been expressed in the heterologous host Escherichia coli, biochemically characterized and used for the kinetic resolution of (rac)-trans-1,2-diacetoxycyclohexane. Both enzymes were selectively acting on the (R,R)-enantiomer of the racemic substrate, highly specifically hydrolyzing only one of the two ester groups present, thus allowing the preparation of enantiopure (R,R)- and (S,S)-cyclohexane-trans-1,2-diol. The reaction conditions for the use of purified enzyme and crude cell lyophilizate were optimized and reactions in batch and repetitive batch modes were carried out on a preparative scale to yield enantiopure product (>99% enantiomeric excess).
来自枯草芽孢杆菌的两种细胞外脂肪酶，即枯草芽孢杆菌脂肪酶A和脂肪酶B，已在异源宿主大肠杆菌中表达，进行了生化表征，并用于（外消旋）-反式-1,2-二乙酰氧基环己烷的动力学拆分。两种酶都选择性地作用于外消旋底物的（R,R）-对映体，高度特异性地仅水解存在的两个酯基中的一个，从而使得能够制备对映体纯的（R,R）-和（S,S）-反式-1,2-环己二醇。优化了使用纯化酶和粗细胞冻干物的反应条件，并在制备规模上以分批和重复分批模式进行反应，以产生对映体纯产物（对映体过量>99%）。
