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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.

DOI:10.1042/bj2260601
PMID:3922354
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1144749/
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。

相似文献

1
Chemical synthesis and papain-catalysed hydrolysis of N-alpha-benzyloxycarbonyl-L-lysine p-nitroanilide.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
Biochem J. 1985 Mar 1;226(2):601-6. doi: 10.1042/bj2260601.
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引用本文的文献

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The preparation and properties of two new chromogenic substrates of trypsin.两种新型胰蛋白酶显色底物的制备与性质
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Cryoenzymology of trypsin. A detailed kinetic study of the trypsin-catalysed hydrolysis of N-alpha-benzyloxycarbonyl-L-lysine p-nitrophenyl ester at low temperatures.胰蛋白酶的低温酶学。对胰蛋白酶催化的N-α-苄氧羰基-L-赖氨酸对硝基苯酯在低温下水解的详细动力学研究。
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