Chaudhry G Rasul, Mateen A, Kaskar B, Bloda M, Riazuddin Sheikh
Department of Biological Sciences, 327 SEB, Oakland University, Rochester, MI 48309-4401, U.S.A.
Biotechnol Appl Biochem. 2002 Aug;36(1):63-70. doi: 10.1042/ba20020026.
A soluble carbamate hydrolase that had a wide specificity was purified 2032-fold from Pseudomonas sp. 50432. This was achieved using a combination of anion-exchange, gel-filtration and hydrophobic-interaction- chromatography techniques. Carbamate hydrolase cleaved the ester linkage of the N-methylcarbamates. The native enzyme was a monomer with a molecular mass of 88 kDa. The optimum pH and temperature of the enzyme activity were 8.5 and 37 degrees C respectively. The tested cations or EDTA did not affect the enzyme activity. However, 2-mercaptoethanol reversibly inhibited the enzyme activity. The enzyme showed the K(m) values of 16 and 12 microM for carbofuran and carbaryl respectively. The purified enzyme did not hydrolyse o-nitrophenyl dimethylcarbamate but hydrolysed several N-methylcarbamates and 1-naphthyl acetate.
从假单胞菌属50432中纯化出一种具有广泛特异性的可溶性氨基甲酸酯水解酶，纯化倍数达2032倍。这是通过阴离子交换、凝胶过滤和疏水相互作用色谱技术相结合实现的。氨基甲酸酯水解酶可裂解N - 甲基氨基甲酸酯的酯键。天然酶为单体，分子量为88 kDa。酶活性的最适pH和温度分别为8.5和37℃。所测试的阳离子或EDTA不影响酶活性。然而，2 - 巯基乙醇可可逆地抑制酶活性。该酶对克百威和西维因的K(m)值分别为16和12 μM。纯化后的酶不水解邻硝基苯基二甲基氨基甲酸酯，但可水解几种N - 甲基氨基甲酸酯和乙酸1 - 萘酯。
