Ogino H, Nakagawa S, Shinya K, Muto T, Fujimura N, Yasuda M, Ishikawa H
Department of Chemical Engineering, Osaka Prefecture University, 1-1 Gakuen-cho, Sakai, Osaka 599-8531, Japan.
J Biosci Bioeng. 2000;89(5):451-7. doi: 10.1016/s1389-1723(00)89095-7.
An organic solvent-stable lipase (LST-03 lipase) secreted into the culture broth of the organic solvent-tolerant Pseudomonas aeruginosa LST-03 was purified by ion-exchange and hydrophobic interaction chromatography in the presence of 2-propanol. The purified enzyme was homogeneous as determined by SDS-PAGE. The molecular mass of the lipase was estimated to be 27.1 kDa by SDS-PAGE and 36 kDa by gel filtration. The optimum pH and temperature were 6.0 and 37 degrees C. LST-03 lipase was stable at pH 5-8 and below 40 degrees C. Its hydrolytic activity was highest against tricaproin (C6), methyl octanoate (C8), and coconut oil respectively among the triacylglycerols, fatty acid methyl esters, and natural oils investigated. The enzyme cleaved not only the 1,3-positioned ester bonds, but also the 2-positioned ester bond of triolein. It exhibited high levels of activity in the presence of n-decane, n-octane, DMSO, and DMF as well as in the absence of an organic solvent. In addition, LST-03 lipase was stabler in the presence of n-decane, ethyleneglycol, DMSO, n-octane, n-heptane, isooctane, and cyclohexane than in the absence of an organic solvent.
一种分泌到耐有机溶剂铜绿假单胞菌LST - 03培养液中的有机溶剂稳定脂肪酶（LST - 03脂肪酶），在2 - 丙醇存在的情况下，通过离子交换和疏水相互作用色谱法进行纯化。经SDS - PAGE测定，纯化后的酶呈均一状态。通过SDS - PAGE估计该脂肪酶的分子量为27.1 kDa，通过凝胶过滤法估计为36 kDa。最适pH值和温度分别为6.0和37℃。LST - 03脂肪酶在pH 5 - 8和40℃以下稳定。在所研究的三酰甘油、脂肪酸甲酯和天然油中，其对三己酸甘油酯（C6）、辛酸甲酯（C8）和椰子油的水解活性分别最高。该酶不仅能切割三油酸甘油酯1,3位的酯键，还能切割其2位的酯键。在正癸烷、正辛烷、二甲基亚砜和二甲基甲酰胺存在以及不存在有机溶剂的情况下，它都表现出较高的活性水平。此外，与不存在有机溶剂时相比，LST - 03脂肪酶在正癸烷、乙二醇、二甲基亚砜、正辛烷、正庚烷、异辛烷和环己烷存在时更稳定。
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