Molecular cloning and sequencing of the phenol hydroxylase gene from Pseudomonas putida BH |
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| Institution: | 1. Department of Applied Chemistry, Himeji Institute of Technology, 2167 Shosha, Himeji, Hyogo 671-22, Japan;2. Department of Environmental Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565, Japan;1. State Key Laboratory of Pollution Control and Resource Reuse, School of the Environment, Nanjing University, Nanjing 210046, PR China;2. State Environmental Protection Engineering Center for Organic Chemical Industrial Waste Water Disposal Resource Reuse, Nanjing 210046, PR China;1. Kompetenzzentrum Holz GmbH, Altenbergerstraße 69, 4040, Linz, Austria;2. Brandenburg Technical University Cottbus-Senftenberg, Germany;1. Graduate School of Environmental and Life Science, Okayama University, 1-1-1 Tsushima-naka, Kita-ku, Okayama 700-8530, Japan;2. Faculty of Science, Japan Women''s University, 2-8-1 Mejirodai, Bunkyo-ku, Tokyo 112-8681, Japan;3. Saitama Gensyu Ikuseikai Co. Ltd, 2616 Niibori, Shobu, Kuki-City, Saitama 346-0105, Japan;4. Faculty of Agriculture, Kochi University, 200 Monobe, Nangoku-City, Kochi 783-8502, Japan;5. Advanced Technology Research Laboratories, Nippon Steel & Sumitomo Metal Corporation, 20-1 Shintomi, Futtsu-City, Chiba 293-8511, Japan;6. Proteo-Science Center, Ehime University, 3 Bunkyo-cho, Matsuyama, Ehime 790-8577, Japan;7. Graduate School of Life and Environmental Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8577, Japan |
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| Abstract: | A genomic library of the phenol-degrading bacterium Pseudomonas putida BH was constructed in the broad host range cosmid pVK100 and introduced into Escherichia coli HB101. One of the recombinant cosmids recovered from catechol- and/or 2-hydroxymuconic semialdehyde-accumulating clones, pS10–45, had a 19.6-kb insert fragment which allowed P. putida KT2440 to grow on phenol as a sole carbon and energy source. Subcloning and expression studies indicated that the phenol hydroxylase gene cluster (pheA) is located on a 6.1-kb SacI fragment. The results of DNA sequencing of the SacI fragment revealed that the pheA gene cluster encodes a multicomponent phenol hydroxylase. |
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