Development of a Rhodococcus recombinant strain for degradation of products from anaerobic dechlorination of PCBs

Environ Sci Technol. 2001 Feb 15;35(4):663-8. doi: 10.1021/es001308t.

Abstract

The Gram-positive bacterium Rhodococcus sp. strain RHA1, naturally containing the biphenyl pathway, was electroporated with a broad host range plasmid containing the 4-chlorobenzoate (4-CBA) degradation operon (fcb) isolated from Arthrobacter globiformis strain KZT1. The recombinant strain grew in medium containing 4-CBA and 4-chlorobiphenyl (4-CB) as the only source of carbon, with stoichiometric release of chloride and a molar growth yield on 4-CB that suggested utilization of both biphenyl rings. In resting cell assays, similar rates of degradation were observed for wild-type and recombinant strains for the most common eight congeners from the anaerobic dechlorination of Aroclor 1242, but the recombinant strain accumulated lower amounts of chlorinated meta-cleavage products and no 4-CBA. Recombinant cells inoculated at 10(4) cells/g into nonsterile soil amended with 4-CB grew to 6-10(5) cells/g, a density consistent with the 4-CB consumed. 4-CB was removed only in the inoculated soil, and the recombinant strain did not grow in the same soil when it was not amended with 4-CB. The fcb operon remained stable in the recombinant strain reisolated from soil after 60 days. This work provides proof of concept that a Rhodococcus strain constructed to grow on a PCB would grow in nonsterile soil if the appropriate chlorobiphenyl is available.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Biodegradation, Environmental
  • Chlorobenzoates / metabolism
  • DNA, Bacterial / genetics
  • Environmental Pollutants / metabolism*
  • Genetic Engineering
  • Operon
  • Plasmids
  • Polychlorinated Biphenyls / metabolism*
  • Rhodococcus / genetics*
  • Rhodococcus / physiology*

Substances

  • Chlorobenzoates
  • DNA, Bacterial
  • Environmental Pollutants
  • Polychlorinated Biphenyls
  • 4-chlorobenzoic acid