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Applied and Environmental Microbiology, January 2003, p. 146-153, Vol. 69, No. 1
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.1.146-153.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Novel Approach to the Improvement of Biphenyl and Polychlorinated Biphenyl Degradation Activity: Promoter Implantation by Homologous Recombination

Yoshiyuki Ohtsubo,1,2* Minoru Shimura,3 Mina Delawary,1 Kazuhide Kimbara,3 Masamichi Takagi,1 Toshiaki Kudo,2 Akinori Ohta,1 and Yuji Nagata4

Department of Biotechnology, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657,1 Molecular Microbial Ecology Division, RIKEN, Hirosawa 2-1, Wako, Saitama 351-0198,2 Railway Technical Research Institute, Kokubunji-shi, Tokyo 185-8540,3 Graduate School of Life Sciences, Tohoku University, Katahira, Sendai 980-8577, Japan4

Received 5 August 2002/ Accepted 28 October 2002

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.

* Corresponding author. Mailing address: Molecular Microbial Ecology Division, RIKEN (The Institute of Physical and Chemical Research), Hirosawa 2-1, Wako, Saitama 351-0198, Japan. Phone: 81-48-467-9544. Fax: 81-48-462-4672. E-mail: yohtsubo{at}postman.riken.go.jp.

Applied and Environmental Microbiology, January 2003, p. 146-153, Vol. 69, No. 1
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.1.146-153.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Villacieros, M., Whelan, C., Mackova, M., Molgaard, J., Sanchez-Contreras, M., Lloret, J., Aguirre de Carcer, D., Oruezabal, R. I., Bolanos, L., Macek, T., Karlson, U., Dowling, D. N., Martin, M., Rivilla, R. (2005). Polychlorinated Biphenyl Rhizoremediation by Pseudomonas fluorescens F113 Derivatives, Using a Sinorhizobium meliloti nod System To Drive bph Gene Expression. Appl. Environ. Microbiol. 71: 2687-2694 [Abstract] [Full Text]  


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