Cloning, sequencing, and overexpression of the Anaerobiospirillum succiniciproducens phosphoenolpyruvate carboxykinase (pckA) gene

被引:38
作者
Laivenieks, M
Vieille, C
Zeikus, JG
机构
[1] MICHIGAN BIOTECHNOL INST, LANSING, MI 48909 USA
[2] MICHIGAN STATE UNIV, DEPT BIOCHEM, E LANSING, MI 48824 USA
关键词
D O I
10.1128/AEM.63.6.2273-2280.1997
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
The phosphoenolpyruvate (PEP) carboxykinase-encoding gene from the anaerobic, CO2-fixing, succinate-producing bacterium Anaerobiospirillum succiniciproducens was cloned, sequenced, and expressed in Escherichia coli. The gene encoded a 532-residue polypeptide with a calculated molecular mass of 58.7 kDa, The sequence of the A. succiniciproducens PEP carboxykinase was similar to those of all known ATP/ADP-dependent PEP carboxykinases. In particular, the A. succiniciproducens enzyme was 67.3% identical and 79.2% similar to the E. coli enzyme, The A. succiniciproducens pckA transcription start site was determined, and putative promoter regions were identified, The recombinant enzyme was overexpressed in E. coli, The purified enzyme was indiscernible from the native enzyme by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and had the same activity as the native enzyme.
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页码:2273 / 2280
页数:8
相关论文
共 32 条
[1]   EVIDENCE FOR A CODING PATTERN ON THE NON-CODING STRAND OF THE ESCHERICHIA-COLI GENOME [J].
ALFFSTEINBERGER, C .
NUCLEIC ACIDS RESEARCH, 1984, 12 (05) :2235-2241
[2]  
Ausubel FM., 1994, Curr. Protoc. Mol. Biol
[3]  
CANNATA JJB, 1974, J BIOL CHEM, V249, P3356
[4]   CONTROL OF GLUCONEOGENIC GROWTH BY PPS AND PCK IN ESCHERICHIA-COLI [J].
CHAO, YP ;
PATNAIK, R ;
ROOF, WD ;
YOUNG, RF ;
LIAO, JC .
JOURNAL OF BACTERIOLOGY, 1993, 175 (21) :6939-6944
[5]   PHOSPHOENOLPYRUVATE CARBOXYKINASE (GUANOSINE 5'-TRIPHOSPHATE) FROM RAT-LIVER CYTOSOL - DUAL-CATION REQUIREMENT FOR THE CARBOXYLATION REACTION [J].
COLOMBO, G ;
CARLSON, GM ;
LARDY, HA .
BIOCHEMISTRY, 1981, 20 (10) :2749-2757
[6]   PHOSPHOENOLPYRUVATE CARBOXYKINASE FROM TRYPANOSOMA-CRUZI - PURIFICATION AND PHYSICOCHEMICAL AND KINETIC-PROPERTIES [J].
CYMERYNG, C ;
CAZZULO, JJ ;
CANNATA, JJB .
MOLECULAR AND BIOCHEMICAL PARASITOLOGY, 1995, 73 (1-2) :91-101
[7]  
DATTA R, 1989, Patent No. 4885247
[8]   A COMPREHENSIVE SET OF SEQUENCE-ANALYSIS PROGRAMS FOR THE VAX [J].
DEVEREUX, J ;
HAEBERLI, P ;
SMITHIES, O .
NUCLEIC ACIDS RESEARCH, 1984, 12 (01) :387-395
[9]   ACTIVITY AND REGULATION OF ANAPLEROTIC AND GLUCONEOGENETIC ENZYMES DURING GROWTH PROCESS OF BAKERS-YEAST - BIPHASIC GROWTH [J].
HAARASILTA, S ;
OURA, E .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1975, 52 (01) :1-7
[10]   2 ROUTES FOR SYNTHESIS OF PHOSPHOENOLPYRUVATE FROM C4-DICARBOXYLIC ACIDS [J].
HANSEN, EJ ;
JUNI, E .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1974, 59 (04) :1204-1210