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 Sprenger, G. A. Articles by Sahm, H. Search for Related Content PubMed PubMed Citation Articles by Sprenger, G. A. Articles by Sahm, H.

 Previous Article  |  Next Article 

J. Bacteriol., 10 1995, 5930-5936, Vol 177, No. 20
Copyright © 1995, American Society for Microbiology

Transaldolase B of Escherichia coli K-12: cloning of its gene, talB, and characterization of the enzyme from recombinant strains

GA Sprenger, U Schorken, G Sprenger and H Sahm
Institut fur Biotechnologie 1, Forschungszentrum Julich GmbH, Julich, Germany.

A previously recognized open reading frame (T. Yura, H. Mori, H. Nagai, T. Nagata, A. Ishihama, N. Fujita, K. Isono, K. Mizobuchi, and A. Nakata, Nucleic Acids Res. 20:3305-3308) from the 0.2-min region of the Escherichia coli K-12 chromosome is shown to encode a functional transaldolase activity. After cloning of the gene onto high-copy-number vectors, transaldolase B (D-sedoheptulose-7-phosphate:D-glyceraldehyde- 3-phosphate dihydroxyacetone transferase; EC 2.2.1.2) was overexpressed up to 12.7 U mg of protein-1 compared with less than 0.1 U mg of protein-1 in wild-type homogenates. The enzyme was purified from recombinant E. coli K-12 cells by successive ammonium sulfate precipitations (45 to 80% and subsequently 55 to 70%) and two anion- exchange chromatography steps (Q-Sepharose FF, Fractogel EMD-DEAE tentacle column; yield, 130 mg of protein from 12 g of cell wet weight) and afforded an apparently homogeneous protein band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis with a subunit size of 35,000 +/- 1,000 Da. As the enzyme had a molecular mass of 70,000 Da by gel filtration, transaldolase B is likely to form a homodimer. N- terminal amino acid sequencing of the protein verified its identity with the product of the cloned gene talB. The specific activity of the purified enzyme determined at 30 degrees C with the substrates fructose- 6-phosphate (donor of C3 compound) and erythrose-4-phosphate (acceptor) at an optimal pH (50 mM glycylglycine [pH 8.5]) was 60 U mg-1.Km values for the substrates fructose-6-phosphate and erythrose-4-phosphate were determined at 1,200 and 90 microM, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

This article has been cited by other articles:

• Schneider, S., Sandalova, T., Schneider, G., Sprenger, G. A., Samland, A. K. (2008). Replacement of a Phenylalanine by a Tyrosine in the Active Site Confers Fructose-6-phosphate Aldolase Activity to the Transaldolase of Escherichia coli and Human Origin. J. Biol. Chem. 283: 30064-30072 [Abstract] [Full Text]  
• Blaudeck, N., Kreutzenbeck, P., Muller, M., Sprenger, G. A., Freudl, R. (2005). Isolation and Characterization of Bifunctional Escherichia coli TatA Mutant Proteins That Allow Efficient Tat-dependent Protein Translocation in the Absence of TatB. J. Biol. Chem. 280: 3426-3432 [Abstract] [Full Text]  
• Blaudeck, N., Kreutzenbeck, P., Freudl, R., Sprenger, G. A. (2003). Genetic Analysis of Pathway Specificity during Posttranslational Protein Translocation across the Escherichia coli Plasma Membrane. J. Bacteriol. 185: 2811-2819 [Abstract] [Full Text]  
• Firoved, A. M., Boucher, J. C., Deretic, V. (2002). Global Genomic Analysis of AlgU ({sigma}E)-Dependent Promoters (Sigmulon) in Pseudomonas aeruginosa and Implications for Inflammatory Processes in Cystic Fibrosis. J. Bacteriol. 184: 1057-1064 [Abstract] [Full Text]  
• Berlyn, M. K. B. (1998). Linkage Map of Escherichia coli K-12, Edition 10: The Traditional Map. Microbiol. Mol. Biol. Rev. 62: 814-984 [Abstract] [Full Text]  
• Schurmann, M., Sprenger, G. A. (2001). Fructose-6-phosphate Aldolase Is a Novel Class I Aldolase from Escherichia coli and Is Related to a Novel Group of Bacterial Transaldolases. J. Biol. Chem. 276: 11055-11061 [Abstract] [Full Text]