High-level expression of maize C-4-type phosphoenolpyruvate carboxylase in Escherichia coli and its rapid purification

被引：13

作者：

Dong, LY ^{[1
]}

Hata, S ^{[1
]}

Izui, K ^{[1
]}

机构：

[1] KYOTO UNIV, FAC AGR, APPL BOT LAB, SAKYO KU, KYOTO 60601, JAPAN

来源：

BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY | 1997年 / 61卷 / 03期

关键词：

phosphoenolpyruvate carboxylase; expression; purification; pET32; plasmid;

D O I：

10.1271/bbb.61.545

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Maize C-4-type phosphoenolpyruvate carboxylase (PEPC) was expressed in E. coli with the pET32 system. The expressed fusion PEPC was active and its amount comprised more than 10% of total soluble protein, The specific activity increased by about 45-fold, compared with our previous system [S. Yanagisawa and K. Izui, Agr ic. Biol, Chem., 54, 241-243 (1990)]. The fusion PEPC was rapidly purified with His bind metal chelation resin, showing a single band on SDS-PAGE, Moreover, the tag domain fused at the N-terminus did not have any effect on catalytic and regulatory properties of PEPC.

引用

页码：545 / 546

页数：2

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