Nakatani T, Hiratake J, Yoshikawa K, Nishioka T, Oda J
a Institute for Chemical Research, Kyoto University , Uji , Kyoto 611 , Japan.
Biosci Biotechnol Biochem. 1992 Jan;56(7):1118-23. doi: 10.1271/bbb.56.1118.
A microbial lipase from Pseudomonas aeruginosa TE3285 was treated in anhydrous diisopropyl ether with three kinds of serine-reactive reagents, ethyl p-nitrophenyl methylphosphonate (ENMP), diisopropyl fluorophosphate (DFP), and phenylmethylsulfonyl fluoride (PMSF) to lose its catalytic activity for both transesterification in an organic solvent and ester hydrolysis in aqueous system. In contrast with the facile inactivation in an organic solvent, no or very slow inactivation was observed in an aqueous solution. The lipase was shown to behave more like a typical serine enzyme in an organic solvent than in aqueous solution with regard to the chemical inactivation by serine-reactive reagents. The unique behavior of the lipase in an organic solvent may be associated with inferfacial activation of the lipase, which is one of the most distinct characteristics of the lipase family, and the activiation of lipase could be induced by a hydrophobic interaction with an organic solvent.
用三种丝氨酸反应试剂，即对硝基苯基甲基膦酸乙酯（ENMP）、二异丙基氟磷酸酯（DFP）和苯甲基磺酰氟（PMSF），在无水异丙醚中处理铜绿假单胞菌TE3285来源的一种微生物脂肪酶，使其失去在有机溶剂中进行酯交换反应以及在水体系中进行酯水解反应的催化活性。与在有机溶剂中容易失活形成对比的是，在水溶液中未观察到失活现象，或者失活非常缓慢。就丝氨酸反应试剂导致的化学失活而言，该脂肪酶在有机溶剂中的表现比在水溶液中更像是一种典型的丝氨酸酶。脂肪酶在有机溶剂中的独特行为可能与脂肪酶的界面激活有关，这是脂肪酶家族最显著的特征之一，而脂肪酶的激活可能是由与有机溶剂的疏水相互作用诱导的。
