The role of thioredoxin and disulfide isomerase in the expression of the snake venom thrombin-like enzyme calobin in Escherichia coli BL21 (DE3)

被引：27

作者：

Yuan, SL ^{[1
]}

Duan, HQ ^{[1
]}

Liu, CJ ^{[1
]}

Liu, XL ^{[1
]}

Liu, TT ^{[1
]}

Tao, HX ^{[1
]}

Zhang, ZS ^{[1
]}

机构：

[1] Beijing Inst Biotechnol, Beijing 100071, Peoples R China

来源：

PROTEIN EXPRESSION AND PURIFICATION | 2004年 / 38卷 / 01期

关键词：

thrombin-like enzyme; thioredoxin; disulfide isomerase; soluble expression; coexpression;

D O I：

10.1016/j.pep.2004.08.004

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Thrombin-like enzymes (TLEs) are studied widely because of their therapeutic potential in myocardial infarction and thrombotic diseases. But they are always produced in Escherichia coli BL21 (DE3) as inclusion bodies because they are single chain Cys-rich proteins. In this work, coexpressing of thioredoxin (TrxA) largely increased the solubility of calobin, one of the TLEs from korea viper Agkistrodon caliginosus, but soluble calobin-T had poor enzyme activity. Disulfide isomerase (DsbC) was introduced into the host cell and coexpressed with calobin in the presence of TrxA, the result was that both the solubility and enzyme activity of calobin-TD were higher than those of calobin-T. Although recombinant calobins exhibited the enzyme of hydrolyzing the fibrinogen, they lost clotting activity to the substrate. Recombinant calobin-TD remained poor amidolytic activity, the effects of divalent metal cations and various inhibitors on this activity were similar to those of native calobin nevertheless, suggesting that calobin-TD exhibited the characteristics of serine protease especially of trypsin-like serine protease. (C) 2004 Elsevier Inc. All rights reserved.

引用

页码：51 / 60

页数：10

共 6 条

[1] Expression of gloshedobin, a thrombin-like enzyme from the venom of Gloydius shedaoensis, in Escherichia coli
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[2] Expression of gloshedobin, a thrombin-like enzyme from the venom of Gloydius shedaoensis, in Escherichia coli
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