Characterization of Alkylphenol Degradation Gene Cluster in Pseudomonas putida MT4 and Evidence of Oxidation of Alkylphenols and Alkylcatechols with Medium-Length Alkyl Chain(ENVIRONMENTAL BIOTECHNOLOGY)
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概要
- 論文の詳細を見る
Alkylphenols (APs) are ubiquitous contaminants in aquatic environments and have endocrine disrupting and toxic effects on aquatic organisms. To investigate biodegradation mechanisms of APs, an AP degradation gene cluster was cloned from a butylphenol (BP)-degrading bacterium, Pseudomonas putida MT4. The gene cluster consisted of 13 genes named bupBA1A2A3A4A5A6CEHIFG From the nucleotide sequences, bupA1A2A3A4A5A6 were predicted to encode a multicomponent phenol hydroxylase (PH), whereas bupBCEHIFG were expected to encode meta-cleavage pathway enzymes. A partial sequence of a putative NtrC-type regulatory gene, bupR, was also found upstream of the gene bupB. This result indicates that APs can be initially oxidized into alkylcatechols (ACs), followed by the meta-cleavage of the aromatic rings. To confirm this pathway, AP degradation tests were carried out using the recombinant P. putida KT2440 harboring the PH genes (bupA1A2A3A4A5A6). The recombinant strain oxidized 4-n-APs with an alkyl chain of up to C7 (≤C7) efficiently and also several BPs including those with an alkyl chain with some degree of branching. Therefore, it was found that PH had a broad substrate specificity for APs with a medium-length alkyl chain (C3-C7). Moreover, the cell extract of a recombinant Escherichia coli harboring bupB (a catechol 2,3-dioxygenase gene) converted 4-n-ACs with an alkyl chain of ≤C9 into yellow meta-cleavage products with a maximum absorbance at 379nm, indicating that the second step enzyme in this pathway is also responsible for the degradation of ACs with a mediumlength alkyl chain. These results suggest that MT4 is a very useful strain in the biodegradation of a wide range of APs with a medium-length alkyl chain, which known nonylphenol-degrading Sphingomonas strains have never degraded.
- 社団法人日本生物工学会の論文
- 2006-10-25
著者
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Takeo Masahiro
Department of Materials Science and Chemistry, Graduate School of Engineering, University of Hyogo
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Kato Dai-ichiro
Department of Materials Science and Chemistry, Graduate School of Engineering, University of Hyogo
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Negoro Seiji
Department of Materials Science and Chemistry, Graduate School of Engineering, University of Hyogo
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Hirai Makoto
Department Of Circulation And Respiration Research Institute Of Environmental Medicine Nagoya Univer
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Negoro Seiji
Department Of Materials Science And Chemistry Graduate School Of Engineering University Of Hyogo
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Negoro Seiji
Department Of Biotechnology Faculty Of Engineering Osaka University
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Takeo Masahiro
Department Of Materials Science And Chemistry Graduate School Of Engineering University Of Hyogo
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Takeo Masahiro
Department Of Applied Chemistry Himeiji Institute Of Technology
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KITAMURA Chitoshi
Department of Materials Science and Chemistry, Graduate School of Engineering, University of Hyogo
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Negoro Seiji
Graduate School Of Engineering Univ. Of Hyogo
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Takahashi Hana
Department of Materials Science and Chemistry, Graduate School of Engineering, University of Hyogo
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Prabu Subbuswamy
Chemicals Assessment Center, Chemicals Evaluation and Research Institute
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Takeo Masahiro
兵庫県立大学 工学研究科物質系工学専攻
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Negoro Seiji
兵庫県立大学 工学研究科物質系工学専攻
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Takahashi Hana
Department Of Materials Science And Chemistry Graduate School Of Engineering University Of Hyogo
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Kitamura Chitoshi
Department Of Materials Science And Chemistry Graduate School Of Engineering Himeji Institute Of Tec
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Kato Dai-ichiro
Graduate School Of Engineering University Of Hyogo
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Kato Dai-ichiro
Department Of Materials Science And Chemistry Graduate School Of Engineering University Of Hyogo
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Hirai Makoto
Department Of Materials Science And Chemistry Graduate School Of Engineering University Of Hyogo
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Prabu Subbuswamy
Chemicals Assessment Center Chemicals Evaluation And Research Institute
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Hirai Makoto
Department of Cardiology, Nagoya University Graduate School of Health Science
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