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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

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.

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