Current Microbiology

, Volume 46, Issue 2, pp 0083–0087

Cloning and Characterization of katA, Encoding the Major Monofunctional Catalase from Xanthomonas campestris pv. phaseoli and Characterization of the Encoded Catalase KatA

  • Nopmanee Chauvatcharin
  • Paiboon Vattanaviboon
  • Jack Switala
  • Peter C. Loewen
  • Skorn Mongkolsuk

DOI: 10.1007/s00284-002-3812-8

Cite this article as:
Chauvatcharin, N., Vattanaviboon, P., Switala, J. et al. Curr Microbiol (2003) 46: 0083. doi:10.1007/s00284-002-3812-8

Abstract

The first cloning and characterization of the gene katA, encoding the major catalase (KatA), from Xanthomonas is reported. A reverse genetic approach using a synthesized katA-specific DNA probe to screen a X. campestris pv. phaseoli genomic library was employed. A positively hybridizing clone designated pKat29 that contained a full-length katA was isolated. Analysis of the nucleotide sequence revealed an open reading frame of 1,521 bp encoding a 507-amino acid protein with a theoretical molecular mass of 56 kDa. The deduced amino acid sequence of KatA revealed 84% and 78% identity to CatF of Pseudomonas syringae and KatB of P. aeruginosa, respectively. Phylogenetic analysis places Xanthomonas katA in the clade I group of bacterial catalases. Unexpectedly, expression of katA in a heterologous Escherichia coli host resulted in a temperature-sensitive expression. The KatA enzyme was purified from an overproducing mutant of X. campestris and was characterized. It has apparent Km and Vmax values of 75 mM [H2O2] and 2.55 × 105 μmol H2O2 μmol heme−1 s−1, respectively. The enzyme is highly sensitive to 3-amino-1,2,4-triazole and NaN3, has a narrower optimal pH range than other catalases, and is more sensitive to heat inactivation.

Copyright information

© Springer-Verlag New York Inc. 2003

Authors and Affiliations

  • Nopmanee Chauvatcharin
    • 2
  • Paiboon Vattanaviboon
    • 1
  • Jack Switala
    • 3
  • Peter C. Loewen
    • 3
  • Skorn Mongkolsuk
    • 1
  1. 1.Laboratory of Biotechnology, Chulabhorn Research Institute, Lak Si, Bangkok 10210, ThailandTH
  2. 2.Department of Biotechnology, Faculty of Science, Mahidol University, Bangkok 10400, ThailandTH
  3. 3.Department of Microbiology, University of Manitoba, Winnipeg, Manitoba, R3T 2N2, CanadaCA

Personalised recommendations