A NEW STRATEGY FOR THE CLONING, OVEREXPRESSION AND ONE-STEP PURIFICATION OF 3 DHAP-DEPENDENT ALDOLASES - RHAMNULOSE-1-PHOSPHATE ALDOLASE, FUCULOSE-1-PHOSPHATE ALDOLASE AND TAGATOSE-1,6-DIPHOSPHATE ALDOLASE

被引：31

作者：

GARCIAJUNCEDA, E ^{[1
]}

SHEN, GJ ^{[1
]}

SUGAI, T ^{[1
]}

WONG, CH ^{[1
]}

机构：

[1] Scripps Res Inst, DEPT CHEM, LA JOLLA, CA 92037 USA

来源：

BIOORGANIC & MEDICINAL CHEMISTRY | 1995年 / 3卷 / 07期

关键词：

D O I：

10.1016/0968-0896(95)00077-T

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：

Three DHAP-dependent aldolases, rhamnulose-1-phosphate aldolase (Rham-1PA), fuculose-1-phosphate aldolase (Fuc-1PA) and tagatose-1,6-diphosphate aldolase (TDPA) have been cloned and overexpressed in Escherichia coli using two different expression vectors: pTrcHis for the expression of Rham-1PA and Fuc-1PA and pRSET for the expression of TDPA. In each case the recombinant enzyme is synthesized as a fusion protein with a hexahistidine tag on the N-terminus. The three enzymes have been purified in only one step by chelation affinity chromatography. The effects of cultivation temperature and concentration of inducer have been studied in order to optimize the expression of the recombinant proteins and to avoid the formation of inclusion bodies.

引用

页码：945 / 953

页数：9

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