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以乙醇为效应物研究对鲍鱼碱性磷酸酶(ALP)活力影响的结果表明,酶的剩余活力随着乙醇浓度增大而迅速下降,乙醇浓度40%可使酶活力完全丧失,说明乙醇对鲍鱼ALP有明显的失活作用,IC50为13%.含较低浓度乙醇(30%)的失活过程是可逆的反应.测定乙醇对酶的失活作用机理,结果表明乙醇对鲍鱼ALP的失活作用是非竞争性机制,说明底物存在不影响乙醇对酶的失活作用.应用荧光光谱、紫外吸收光谱研究鲍鱼ALP经乙醇微扰后的分子构象变化,发现乙醇对酶分子构象有显著的影响,酶的内源荧光强度随乙醇浓度增大而增强,荧光发射峰逐渐发生红移.紫外吸收光谱在276 nm吸收峰随乙醇浓度增大而增强.这些结果表明,酶蛋白分子中的生色基团残基的微环境发生变化.
Ethanol as an effector on abalone alkaline phosphatase (ALP) activity of the results showed that the residual activity of the enzyme with the ethanol concentration decreased rapidly, the concentration of ethanol can completely lose the activity of 40%, indicating that ethanol on abalone ALP has obvious inactivation, with an IC50 of 13% .The inactivation process with lower concentration of ethanol (30%) is a reversible reaction.Determination of ethanol inactivation mechanism, the results showed that ethanol on abalone ALP inactivation The mechanism of action is a non-competitive mechanism, indicating that the presence of substrate does not affect the inactivation of ethanol on the enzyme.Using fluorescence spectroscopy, UV absorption spectroscopy abalone ALP ethanol ethanol after perturbation of the molecular conformational changes found that ethanol on the enzyme molecular conformation significant effect , The endogenous fluorescence intensity of the enzyme increased with the increase of ethanol concentration and the fluorescence emission peak gradually red shifted.The UV absorption peak at 276 nm increased with the increase of ethanol concentration.These results showed that the color of the enzyme protein molecules Microenvironment of radical residues changes.