Novel approach to the improvement of biphenyl and polychlorinated biphenyl degradation activity: promoter implantation by homologous recombination.
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.
Ohtsubo
Yoshiyuki
Y
Department of Biotechnology, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan. yohtsubo@postman.riken.go.jp
Shimura
Minoru
M
Delawary
Mina
M
Kimbara
Kazuhide
K
Takagi
Masamichi
M
Kudo
Toshiaki
T
Ohta
Akinori
A
Nagata
Yuji
Y
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001673:Biodegradation, Environmental; D001713:Biphenyl Compounds / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D005818:Genetic Engineering / Q000379:methods; D006867:Hydrolases / Q000235:genetics / Q000378:metabolism; D009876:Operon; D011078:Polychlorinated Biphenyls / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D011549:Pseudomonas / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D011995:Recombination, Genetic; D014158:Transcription, Genetic
D001426:Bacterial Proteins; D001713:Biphenyl Compounds; D006867:Hydrolases; D011078:Polychlorinated Biphenyls; C010574:biphenyl; C062907:2,6-dioxo-6-phenylhexa-3-enoate hydrolase