Plant Molecular Biology

, Volume 22, Issue 6, pp 937–943

Characterization of pea chloroplastic carbonic anhydrase. Expression inEscherichia coli and site-directed mutagenesis

  • Nicholas J. Provart
  • Nathalie Majeau
  • John R. Coleman
Research Articles

DOI: 10.1007/BF00028967

Cite this article as:
Provart, N.J., Majeau, N. & Coleman, J.R. Plant Mol Biol (1993) 22: 937. doi:10.1007/BF00028967

Abstract

A cDNA encoding the mature, chloroplast-localized carbonic anhydrase in pea has been expressed inE. coli. The enzyme is fully active and yields of up to 20% of the total soluble protein can be obtained from the bacteria. This expression system was used to monitor the effects of site-directed mutagenesis of seven residues found within conserved regions in the pea carbonic anhydrase amino acid sequence. The effects of these modifications are discussed with respect to the potential of various amino acids to act as sites for zinc coordination or intramolecular proton shuttles.

Key words

carbonic anhydraseEscherichia colimutagenesisPisum sativum

Copyright information

© Kluwer Academic Publishers 1993

Authors and Affiliations

  • Nicholas J. Provart
    • 1
  • Nathalie Majeau
    • 1
  • John R. Coleman
    • 1
  1. 1.Dept. of BotanyUniversity of TorontoTorontoCanada