Cloning, biochemical properties, and distribution of mycobacterial haloalkane dehalogenases

Andrea Jesenská, Martina Pavlová, Michal Strouhal, Radka Chaloupková, Iva Těšínská, Marta Monincová, Zbyněk Prokop, Milan Bartoš, Ivo Pavlík, Ivan Rychlík, Petra Möbius, Yuji Nagata, Jiři Damborský

Research output: Contribution to journalArticlepeer-review

40 Citations (Scopus)

Abstract

Haloalkane dehalogenases are enzymes that catalyze the cleavage of the carbon-halogen bond by a hydrolytic mechanism. Genomes of Mycobacterium tuberculosis and M. bovis contain at least two open reading frames coding for the polypeptides showing a high sequence similarity with biochemically characterized haloalkane dehalogenases. We describe here the cloning of the haloalkane dehalogenase genes dmbA and dmbB from M. bovis 5033/66 and demonstrate the dehalogenase activity of their translation products. Both of these genes are widely distributed among species of the M. tuberculosis complex, including M. bovis, M. bovis BCG, M. africanum, M. caprae, M. microti, and M. pinnipedii, as shown by the PCR screening of 48 isolates from various hosts. DmbA and DmbB proteins were heterologously expressed in Escherichia coli and purified to homogeneity. The DmbB protein had to be expressed in a fusion with thioredoxin to obtain a soluble protein sample. The temperature optimum of DmbA and DmbB proteins determined with 1,2-dibromoethane is 45°C. The melting temperature assessed by circular dichroism spectroscopy of DmbA is 47°C and DmbB is 57°C. The pH optimum of DmbA depends on composition of a buffer with maximal activity at 9.0. DmbB had a single pH optimum at pH 6.5. Mycobacteria are currently the only genus known to carry more than one haloalkane dehalogenase gene, although putative haloalkane dehalogenases can be inferred in more then 20 different bacterial species by comparative genomics. The evolution and distribution of haloalkane dehalogenases among mycobacteria is discussed.

Original languageEnglish
Pages (from-to)6736-6745
Number of pages10
JournalApplied and environmental microbiology
Volume71
Issue number11
DOIs
Publication statusPublished - 2005 Nov

ASJC Scopus subject areas

  • Biotechnology
  • Food Science
  • Applied Microbiology and Biotechnology
  • Ecology

Fingerprint Dive into the research topics of 'Cloning, biochemical properties, and distribution of mycobacterial haloalkane dehalogenases'. Together they form a unique fingerprint.

Cite this