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酵母乙醇脱氢酶在十二烷基硫酸钠水溶液中催化乙醇氧化的动力学

Kinetics of ethanol oxidation catalyzed by yeast alcohol dehydrogenase in aqueous solutions of sodium dodecylsulfate.

作者信息

Abuin Elsa, Lissi Eduardo, León Luis

机构信息

Facultad de Química y Biología, Universidad de Santiago de Chile (USACH), Casilla 40-Correo 33, Santiago, Chile.

出版信息

Protein J. 2008 Jun;27(4):247-52. doi: 10.1007/s10930-008-9131-6.

Abstract

A study has been made of the effect of sodium dodecylsufate (SDS) addition on the oxidation of ethanol catalyzed by yeast alcohol dehydrogenase. Experiments were performed at pH = 8.1 and SDS concentrations employed were below and above the surfactant critical micelle concentration (CMC). The double reciprocal plots obtained in the absence and in the presence of the surfactant were compatible with a sequential bi-bi ordered mechanism. In the presence of the surfactant the initial reaction rates were consistently lower than in pure buffer at all the surfactant concentrations considered (0.5-50 mM). This effect is mainly due to an increase in the dissociation constant of beta-NAD(+) which reaches its maximum value (7,100 +/- 1,700 microM) at the CMC. Above the CMC the effect of the surfactant is mainly due to an increase in the Michaels constants of the alcohol, with values of 41 +/- 1 mM for 15 mM SDS and 50 +/- 1 mM for 50 mM SDS. The catalytic rate constant was found to be practically independent of the presence of the surfactant in the range of concentrations considered (up to 50 mM).

摘要

研究了添加十二烷基硫酸钠(SDS)对酵母乙醇脱氢酶催化乙醇氧化的影响。实验在pH = 8.1条件下进行,所用SDS浓度低于和高于表面活性剂临界胶束浓度(CMC)。在不存在和存在表面活性剂的情况下获得的双倒数图符合有序的双底物双产物序列机制。在表面活性剂存在下,在所考虑的所有表面活性剂浓度(0.5 - 50 mM)下,初始反应速率始终低于纯缓冲液中的反应速率。这种效应主要是由于β - NAD⁺解离常数增加,在CMC时达到最大值(7100 ± 1700 μM)。在CMC以上,表面活性剂的影响主要是由于乙醇的米氏常数增加,对于15 mM SDS,其值为41 ± 1 mM,对于50 mM SDS,其值为50 ± 1 mM。发现在所考虑的浓度范围内(高达50 mM),催化速率常数实际上与表面活性剂的存在无关。

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