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
收藏
页码：217 / 225
页数：9
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