Genes in PHT Plasmid Encoding the Initial Degradation Pathway of Phthalate in Pseudomonas putida
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概要
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The nucleotide sequence of a 7-kbp EcoRI fragment of PHT plasmid in Pseudomonas putida encoding the enzyme in the initial step of phthalate degradation (the Pht^+ phenotype) revealed five open reading frames (ORFs), pht1 to pht5,with the same orientation. The protein deduced from pht1 showed 17.4% similarity in amino acid sequence with the glycerol-3-phosphate transporter of Escherichia coli, that of pht2 showed 39.2% similarity with VanB protein (vanillate monooxygenase reductase) and that of pht3 showed 20.8% similarity with VanA (vanillate monooxygenase) of Pseudomonas sp. The proteins encoded by pht4 and pht5 showed no appreciable similarity with any proteins in a database. Eight Pht^- mutants with a Tn5 insertion in pht1,and one with a similar insertion in the pht5 ORF were isolated. These pht1^- and pht5^- mutants, and two other Pht^- mutants induced by UV irradiation of a Pht^+ strain of Pseudomonas testosteroni, were examined by complementation tests with several segments of the 7-kbp pht fragment and feeding experiments with phthalate, 4,5-dihydro-4,5-dihydroxyphthalate, 4,5-dihydroxyphthalate, and protocatechuate. The results of these experiments and similarity searches suggested that pht2 encodes phthalate oxygenase reductase, pht3 encodes phthalate oxygenase, pht4 encodes 4,5-dihydro-4,5-dihydroxyphthalate dehydrogenase, and pht5 encodes 4,5-dihydroxyphthalate decarboxylase. The Pht1 protein was suggested to function as a positive regulator for expression of the other pht genes or as a phthalate transporter. Three A : T rich regions, which may have promoter function, were found in the upstream regions of pht1,pht3,and pht5. Northern blot analyses suggested that transcriptions of pht1 and pht2 are low level and under relaxed control of phthalate. The transcriptions of pht3,pht4,and pht5 might be induced by phthalate.
- 社団法人日本生物工学会の論文
- 1992-12-25
著者
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Harashima Satoshi
Department of Biotechnology, Graduate School of Engineering, Osaka University
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Nakagawa Masato
Graduate School Of Science And Technology Kobe University
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Harashima Satoshi
Department Of Biotechnology Faculty Of Engineering Osaka University
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OGAWA Nobuo
Department of Internal Medicine, Kanagawa Prefectural Respiratory and Cardiovascular Disease Center
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Nakagawa M
Graduate School Of Science And Technology Kobe University
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NOMURA YASUTOSHI
Research Center, Japan Synthetic Rubber Co. Ltd.,
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NAKAGAWA MARIKO
Research Center, Japan Synthetic Rubber Co. Ltd.,
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OSHIMA YASUJI
Department of Biotechnology, Faculty of Engineering, Osaka University
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Ogawa Nobuo
Department Of Biotechnology Faculty Of Engineering Osaka University
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Ogawa Nobuo
Department Of Mechanical Engineering Faculty Of Science And Technology Science University Of Tokyo
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Nomura Yasuo
The Faculty Of Engineering Kansai University
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Oshima Y
Faculty Of Engineering Kansai University
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Oshima Yasuji
Department Of Biotechnology Faculty Of Engineering Osaka University
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Oshima Yasuji
Department Of Biotechnology Faculty Of Engineering Kansai University
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HARASHIMA Satoshi
Department of Biotechnoiogy, Graduate School of Engineering, Osaka University
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