Purification and Characterization of Dibenzothiophene Sulfone Monooxygenase and FMN-Dependent NADH Oxidoreductase from the Thermophilic Bacterium Paenibacillus sp.Strain A11-2
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概要
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A dibenzothiophene(DBT) sulfone monooxygenase(TdsA), which catalyses the oxidative C-S bond cleavage of DBT sulfone to produce 2-(2-hydroxyphenyl)benzenesulfinate(HPBS) was purified from the thermophilic DBT desulfurizing bacterium Paenibacillus sp.strain A11-2 by multistep chromatography. The molecular mass of the purified enzyme was determined to be 120 kDa by gel filtration and the subunit molecular mass was calculated to be 45 kDa by SDS-polyacrylamide gel electrophoresis(SDS-PAGE) indicating a dimeric structure. The N-terminal amino acid sequence of the purified TdsA was determined to be MRQMHLAGFFAAGNTHH, which revealed no significant similarity to any other known amino acid sequences. The purified TdsA absolutely required an oxidoreductase for its activity. This oxidoreductase(TdsD) was also purified to homogeneity, and its molecular size was calculated to be 50 kDa and 25 kDa by gel filtration and SDS-PAGE, respectively. TdsD was completely FMN-dependent, and FAD could not act as a cofactor. The N-terminal amino acid sequence of the purified TdsD was determined to be TSQTAEQSIAPIVAQYRHPEQPISALFVNR, which showed significant similarity to kinesin-like protein(44% identity). The optimal temperatures for the activity of TdsA and TdsD were 45℃ and 55℃, respectively. Both enzymes showed optimal activity at pH 5.5. TdsA slightly inhibited by sulfate, but not by 2-hydroxybiphenyl (2-HBP), which is another and product of DBT. TdsA showed higher activity toward bulkjer substrates than its mesophilic counterpart, DszA. These properties suggest the applicability of biodesulfurization to the processing of actual petroleum fractions.
- 社団法人日本生物工学会の論文
- 2000-12-25
著者
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ISHII YOSHITAKA
Bio-Refining Process Laboratory, Japan Cooperation Center, Petroleum
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MARUHASHI KENJI
Bio-Refining Process Laboratory, Japan Cooperation Center, Petroleum
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SUZUKI MASANORI
Bio-Refining Process Laboratory, Advanced Technology and Research Institute, Petroleum Energy Center
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Ohta Yoshinori
Bio-refining Process Laboratory Advanced Technology And Research Institute Petroleum Energy Center
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Konishi Jin
Bio-refining Process Laboratory Advanced Technology And Research Institute Petroleum Energy Center
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Maruhashi Kenji
Bio-refining Process Laboratory Technical Cooperation Department Japan Cooperation Center
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ONAKA Toshimitsu
Bio-Refining Process Laboratory, Advanced Technology and Research Institute, Petroleum Energy Center
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Suzuki M
Bio-refining Process Laboratory Advanced Technology And Research Institute Petroleum Energy Center
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Suzuki Masanori
Bio-refining Process Laboratory Advanced Technology And Research Institute Petroleum Energy Center
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Ishii Yoshitaka
Bio-refining Process Laboratory Advanced Technology And Research Institute Petroleum Energy Center
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Onaka Toshimitsu
Bio-refining Process Laboratory Advanced Technology And Research Institute Petroleum Energy Center
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