Enzymatic activity assay of D-hydantoinase by isothermal titration calorimetry.: Determination of the thermodynamic activation parameters for the hydrolysis of several substrates

被引：7

作者：

Andújar-Sánchez, M ^{[1
]}

Las Heras-Vázquez, FJ ^{[1
]}

Clemente-Jiménez, JM ^{[1
]}

Martínez-Rodríguez, S ^{[1
]}

Camara-Artigas, A ^{[1
]}

Rodríguez-Vico, F ^{[1
]}

Jara-Pérez, V ^{[1
]}

机构：

[1] Univ Almeria, Dpto Quim Fis Bioquim & Quim Inorgan, Almeria 04120, Spain

来源：

JOURNAL OF BIOCHEMICAL AND BIOPHYSICAL METHODS | 2006年 / 67卷 / 01期

关键词：

D-hydantoinase; isothermal titration calorimetry; activity assay; activation parameters;

D O I：

10.1016/j.jbbm.2006.01.002

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Isothermal titration calorimetry (ITC) has been applied to the determination of the activity of D-hydantoinase (EC 3.5.2.2) with several substrates by monitoring the heat released during the reaction. The method is based on the proportionality between the reaction rate and the thermal power (heat/time) generated. Microcalorimetric assays carried out at different temperatures provided the dependence of the catalytic rate constant on temperature. We show that ITC assay is a nondestructive method that allows the determination of the catalytic rate constant (k(cat)), Michaelis constant (K-M), activation energy and activation Gibbs energy, enthalpy and entropy of this reaction. (c) 2006 Elsevier B.V. All rights reserved.

引用

页码：57 / 66

页数：10

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