Characterization of Phosphoenolpyruvate Carboxylase from Oceanimonas smirnovii in Escherichia coli

被引：4

作者：

Park, Soohyun ^{[1
]}

Lee, Wangjun ^{[1
]}

Kim, Hyeonsoo ^{[1
]}

Pack, Seung Pil ^{[2
]}

Lee, Jinwon ^{[1
]}

机构：

[1] Sogang Univ, Dept Chem & Biomol Engn, Seoul 121742, South Korea

[2] Korea Univ, Dept Biotechnol & Bioinformat, Sejong 339700, South Korea

来源：

APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY | 2015年 / 177卷 / 01期

基金：

新加坡国家研究基金会;

关键词：

Oceanimonas smirnovii; Phosphoenolpyruvate carboxylase; Characterization; Bicarbonate; CARBONIC-ANHYDRASE; EXPRESSION; ENZYMES; GENE;

D O I：

10.1007/s12010-015-1739-3

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：

In this study, phosphoenolpyruvate carboxylase (PEPC) derived from Oceanimonas smirnovii (OS) was expressed as a soluble protein in Escherichia coli BL21(DE3). We isolated OS-PEPC (a recombinant PEPC protein) by his-tag purification. The purified protein showed a single band upon analysis with SDS-PAGE, and it had an apparent molecular mass of 98 kDa. Pufied OS-PEPC showed a specific activity value of 21.8 +/- A 0.495 U/mg protein. Especially, OS-PEPC showed the enzymatic activity between 40 and 50 A degrees C. It maintained enzymatic activity in basic pH conditions (pH value, 9-10). We also measured OS-PEPC PEP and HCO3 (-) saturation kinetics and confirmed the effect of divalent cation on OS-PEPC activity.

引用

页码：217 / 225

页数：9

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