A non-dechlorinating strain of Dehalospirillum multivorans: evidence for a key role of the corrinoid cofactor in the synthesis of an active tetrachloroethene dehalogenase

Arch Microbiol. 2002 Dec;178(6):443-9. doi: 10.1007/s00203-002-0473-8. Epub 2002 Sep 11.

Abstract

A strain of Dehalosprillum multivorans, designated strain N, was isolated from the same source as the formerly described tetrachloroethene (PCE)-dechlorinating D. multivorans, herein after referred to as strain K. Neither growing cells nor cell extracts of strain N were able to dechlorinate PCE. The pceA and pceB genes encoding for the PCE-reductive dehalogenase were detected in cells of strain N; and they were 100% homologous to the corresponding genes of strain K. Since the PCE dehalogenase of D. multivorans strain K contains a corrinoid cofactor, the corrinoids of strain N cells were extracted. Analysis of the corrinoids revealed the absence of the specific corrinoid, which is the cofactor of the PCE dehalogenase of strain K cells. RT-PCR of mRNA indicated that the pceA gene was transcribed in strain N cells to a far lower extent than the pceA gene of strain K under the same experimental conditions. Western blot analysis of crude extracts of strain N showed that, if at all, an insignificant amount of the apoprotein of the PCE dehalogenase was present. The results indicate that the inability of strain N to dechlorinate is due to the absence of the corrinoid cofactor of the enzyme mediating PCE dechlorination.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatography, High Pressure Liquid
  • Corrinoids
  • Epsilonproteobacteria / enzymology*
  • Epsilonproteobacteria / genetics
  • Epsilonproteobacteria / metabolism
  • Molecular Sequence Data
  • Oxidoreductases / metabolism*
  • Porphyrins / physiology*
  • Reverse Transcriptase Polymerase Chain Reaction

Substances

  • Corrinoids
  • Porphyrins
  • Oxidoreductases
  • tetrachloroethene dehalogenase

Associated data

  • GENBANK/AF524868