This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Laivenieks, M. Articles by Zeikus, J. G. Search for Related Content PubMed PubMed Citation Articles by Laivenieks, M. Articles by Zeikus, J. G. Agricola Articles by Laivenieks, M. Articles by Zeikus, J. G.

 Previous Article  |  Next Article 

Appl. Environ. Microbiol., 06 1997, 2273-2280, Vol 63, No. 6
Copyright © 1997, American Society for Microbiology

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

M Laivenieks, C Vieille and JG Zeikus
Department of Biochemistry, Michigan State University, East Lansing 48824, USA.

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.

This article has been cited by other articles:

• Kim, P., Laivenieks, M., Vieille, C., Zeikus, J. G. (2004). Effect of Overexpression of Actinobacillus succinogenes Phosphoenolpyruvate Carboxykinase on Succinate Production in Escherichia coli. Appl. Environ. Microbiol. 70: 1238-1241 [Abstract] [Full Text]  
• Asanuma, N., Hino, T. (2001). Molecular characterization, enzyme properties and transcriptional regulation of phosphoenolpyruvate carboxykinase and pyruvate kinase in a ruminal bacterium, Selenomonas ruminantium. Microbiology 147: 681-690 [Abstract] [Full Text]