僧帽牡蛎碱性磷酸酶在变性剂作用下构象与催化活力变化的研究

Abstract

从僧帽牡蛎（OSTrEACuCullATA）整体中提取一种经聚丙烯酸胺凝胶电泳鉴定为均一纯的碱性磷酸酶。经不同浓度的胍和脲处理,以荧光光谱为监测手段。研究其变性和大活动力学过程,发现酶的荧光强度随着变性剂浓度的增大而逐渐下降,330.2nM处的吸收峰峰值位置也出现了一定程度的“红移”和“蓝”。测定该酶的失活和变性速度常数发现,不论该酶在胍或脲溶液中变性,其失活速度常数都较其变性速度常数大。呈现出“快失活慢构象变化”的过程。

An Alkaline Phosphatase (AK.P) exhibiting polyacrylamide gel electrophoretic homogeneity was obtained From the Ostrea Cncnllata.The studies of denaturation and inactivation kinetics of enzymeby guanidine and urea with Fluorescence method Found that the bigger denaturation chemicals concen-tration, the less Fluorescence absorbance.The position of 330.2 nm absorbance peak showed some ''displacement to blue or to red" determination of the denaturation and inactivation rate constants showed that the inactivation rate constants always bigger than denaturation ones either in guanidine or urea solution.Showing the process of the change of ''Fast inactivation and slow conFormation