N-α-苄氧羰基-L-赖氨酸对硝基苯胺的化学合成及木瓜蛋白酶催化水解
Chemical synthesis and papain-catalysed hydrolysis of N-alpha-benzyloxycarbonyl-L-lysine p-nitroanilide.
作者信息
Mackenzie N E, Malthouse J P, Scott A I
出版信息
Biochem J. 1985 Mar 1;226(2):601-6. doi: 10.1042/bj2260601.
Abstract
The chemical synthesis of N-alpha-benzyloxycarbonyl-L-lysine p-nitroanilide (Z-Lys-pNA) is described in detail. The pH-dependence of the catalytic parameters kcat,' Km and kcat./Km for the papain-catalysed hydrolysis of Z-Lys-pNA are determined. kcat. and Km are pH-independent between pH 5 and pH 7.42, but the pH-dependence of kcat./Km is bell-shaped, decreasing at high and low pH values with pKa values of 7.97 and 4.40 respectively. The catalytic parameters and their pH-dependence are shown to be similar to those reported for other anilide substrates and it is concluded that the Km value of 0.01 mM previously reported [Angelides & Fink (1979) Biochemistry 18, 2355-2369] is incorrect. The possibility of accumulating a tetrahedral intermediate during the papain-catalysed hydrolysis of Z-Lys-pNA is discussed.
摘要
详细描述了N-α-苄氧羰基-L-赖氨酸对硝基苯胺（Z-Lys-pNA）的化学合成。测定了木瓜蛋白酶催化Z-Lys-pNA水解的催化参数kcat、Km和kcat/Km的pH依赖性。在pH 5至pH 7.42之间，kcat和Km与pH无关，但kcat/Km的pH依赖性呈钟形，在高pH值和低pH值时分别以7.97和4.40的pKa值降低。结果表明，催化参数及其pH依赖性与其他苯胺底物报道的相似，得出先前报道的0.01 mM的Km值[Angelides & Fink（1979年）《生物化学》18, 2355 - 2369]是不正确的结论。讨论了在木瓜蛋白酶催化Z-Lys-pNA水解过程中积累四面体中间体的可能性。
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