Novel approach to the improvement of biphenyl and polychlorinated biphenyl degradation activity: promoter implantation by homologous recombination

Appl Environ Microbiol. 2003 Jan;69(1):146-53. doi: 10.1128/AEM.69.1.146-153.2003.

Abstract

To improve the capabilities of microorganisms relevant for biodegradation, we developed a new genetic approach and applied it to the bph operon (bphEGF[orf4]A1A2A3CD[orf1]A4R) of Pseudomonas sp. strain KKS102 to enhance its biphenyl- and polychlorinated biphenyl (PCB)-degrading activity. A native promoter of the bph operon, which was under control, was replaced through homologous recombination by a series of promoters that had constitutive activity. By testing a series of promoters with various strengths, we were able to obtain strains that have enhanced degradation activity for biphenyl and PCBs. This strategy removes the rate-limiting factor associated with transcription and has the potential to improve the degradation activity of a wide variety of microorganisms involved in biodegradation.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Biodegradation, Environmental
  • Biphenyl Compounds / metabolism*
  • Gene Expression Regulation, Bacterial
  • Genetic Engineering / methods
  • Hydrolases / genetics
  • Hydrolases / metabolism
  • Operon
  • Polychlorinated Biphenyls / metabolism*
  • Promoter Regions, Genetic*
  • Pseudomonas / genetics
  • Pseudomonas / growth & development
  • Pseudomonas / metabolism*
  • Recombination, Genetic*
  • Transcription, Genetic

Substances

  • Bacterial Proteins
  • Biphenyl Compounds
  • diphenyl
  • Polychlorinated Biphenyls
  • Hydrolases
  • 2,6-dioxo-6-phenylhexa-3-enoate hydrolase