Kinetic study of thermal inactivation for native and methoxypolyethylene glycol modified trypsin

被引：26

作者：

He, ZM ^{[1
]}

Zhang, ZD ^{[1
]}

He, MX ^{[1
]}

机构：

[1] Tianjin Univ, Enzyme Technol Lab, Chem Engn Res Ctr, Tianjin 300072, Peoples R China

来源：

PROCESS BIOCHEMISTRY | 2000年 / 35卷 / 10期

基金：

中国国家自然科学基金;

关键词：

trypsin; chemical modification; thermal inactivation; kinetic model; autolysis;

D O I：

10.1016/S0032-9592(00)00168-0

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 [生物化学与分子生物学]; 081704 [应用化学];

摘要：

Bovine pancreatic trypsin (Ti) has been modified with four kinds of methoxypolyethylene glycol (MPEG, molecular masses 350, 750, 2000 and 5000 Da) to enhance thermostability. The MPEG-modified Ti was more stable against temperature than the native form, the larger molecular mass moiety of MPEG showing higher thermostability. To investigate the mechanism of thermal inactivation, a new kinetic model, which has the ability of taking the thermal denaturation and autolysis effects of the proteases into account, has been used to analyze the thermal inactivation process of the native and modified Ti in detail. The kinetic analysis showed that the stabilization effect caused by MPEG modification was the result of a decrease in autolysis rate and a decrease in the rate of thermal denaturation. In addition, the possible mechanism of reduced autolysis and lower thermal denaturation rate were also discussed. (C) 2000 Elsevier Science Ltd. All rights reserved.

引用

页码：1235 / 1240

页数：6

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