Mutagenic analysis of the conserved residues in dehalogenase IVa of Burkholderia cepacia MBA4

FEMS Microbiol Lett. 2001 Oct 16;204(1):135-40. doi: 10.1111/j.1574-6968.2001.tb10876.x.

Abstract

Amino and carboxyl terminal deletion derivatives of dehalogenase IVa (DehIVa) of Burkholderia cepacia MBA4 were constructed and analyzed for enzyme activity and for protein integrity. The results suggested that the majority of the protein is indispensable. Point mutations on 29 conserved charged and/or polar residues were generated and characterized. Derivatives D11E, D11N, D11S and D181N were totally inactive while mutant N178D was defective in catalysis. Mutations of other conserved residues displayed varying effects. Mutation that enhances DehIVa activity has been shown to be inhibitory in other dehalogenase and essential conserved residues in DehIVa have been shown to be dispensable in others. This suggests there is no general rule for the importance of these conserved residues.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blotting, Western
  • Burkholderia cepacia / enzymology*
  • Burkholderia cepacia / genetics
  • Burkholderia cepacia / growth & development
  • Gene Deletion*
  • Genes, Bacterial
  • Hydrolases / chemistry*
  • Hydrolases / genetics*
  • Hydrolases / metabolism
  • Point Mutation*

Substances

  • Hydrolases
  • 2-haloacid dehalogenase