Cloning and Sequence Analysis of Hydroxyquinol 1, 2-Dioxygenase Gene in 2, 4, 6-Trichlorophenol-Degrading Ralstonia pickettii DTP0602 and Characterization of Its Product
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概要
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A gene encoding hydroxyquinol 1, 2-dioxygenase was cloned from 2, 4, 6-trichlorophenol-degrading Ralstonia (Pseudomonas) pickettii strain DTP0602. Cell-free extracts of Escherichia coli containing a cloned 1.4-kb StuI-XhoI DNA fragment of R.pickettii DTP0602 hydroxyquinol 1, 2-dioxygenase converted hydroxyquinol into maleylacetate and also degraded 6-chlorohydroxyquinol. The 1.4-kb DNA fragment contained one open reading frame (designated hadC) composed of 948 nucleotides. The molecular mass of 34, 591 deduced from the gene product (HadC) was in agreement with the size (35 kDa) of the purified HadC protein determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The amino acid sequence of HadC exhibited high homology to that of the hydroxyquinol 1, 2-dioxygenase of 2, 4, 5-trichlorophenoxyacetic acid-degrading Burkholderia cepacia AC1100 (Daubaras, D.L.et al., Appl.Environ.Microbiol., 61, 1279-1289, 1995). The active enzyme had a molecular mass of 68kDa, suggesting that it is functional as a homodimer. The enzyme also catalyzed the oxidation of pyrogallol and 3-methylcatechol, possible intermediates in the degradation of 2, 4, 6-trichlorophenol, in addition to 6-chlorohydroxyquinol and hydroxyquinol. The dioxygenase catalyzed both ortho- and meta-cleavage of 3-methylcatechol.
- 社団法人日本生物工学会の論文
- 1999-03-25
著者
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TAKIZAWA Noboru
Department of Pediatrics, Kanazawa University School of Medicine
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NAKANO Osamu
Department of Urology Tohoku University Graduate School of Medicine, Sendai
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KIYOHARA Hohzoh
Department of Applied Chemistry, Faculty of Engineering, Okayama University of Science
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HATTA TAKASHI
Research Institute of Technology, Okayama University of Science
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Hatta T
Okayama Univ. Sci. Okayama Jpn
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Hatta Takashi
Research Institute Of Technology Okayama University Of Science
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Hirose Toru
Department Of Biological Function And Genetic Resources Science Faculty Of Agriculture Okayama Unive
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Imai Nobuyuki
Department Of Pathology Niigata University School Of Dentistry
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Imai Nobuyuki
Department Of Applied Chemistry Faculty Of Engineering Okayama University Of Science
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Takizawa N
Department Of Applied Chemistry Faculty Of Engineering Okayama University Of Science
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Takizawa Noboru
Department Of Pediatrics Kanazawa University School Of Medicine
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Takizawa Noboru
Department Of Applied Chemistry Faculty Of Engineering Okayama University Of Science
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Kiyohara H
Department Of Applied Chemistry Faculty Of Engineering Okayama University Of Science
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Kiyohara Hohzoh
Department Of Applied Chemistry Faculty Of Engineering Okayama University Of Science
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Nakano O
Department Of Urology Tohoku University Graduate School Of Medicine Sendai
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Nakano Osamu
Department Of Pediatrics:hospital Pharmacy:and Faculty Of Pharmaceutical Sciences University Of Kana
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