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Chlorophenol 4-monooxygenase is a key enzyme in the degradation of 2,4,5-trichlorophenoxyacetic acid, and it was originally characterized as a two-component enzyme (TftC and TftD). Sequence analysis suggests that they are separate enzymes. The two proteins were separately produced in\n            <jats:italic>Escherichia coli<\/jats:italic>\n            , purified, and characterized. TftC was an NADH:flavin adenine dinucleotide (FAD) oxidoreductase. A C-terminally His-tagged fusion TftC used NADH to reduce either FAD or flavin mononucleotide (FMN) but did not use NADPH or riboflavin as a substrate. Kinetic and binding property analysis showed that FAD was a better substrate than FMN. TftD was a reduced FAD (FADH\n            <jats:sub>2<\/jats:sub>\n            )-utilizing monooxygenase, and FADH\n            <jats:sub>2<\/jats:sub>\n            was supplied by TftC. It converted 2,4,5-trichlorophenol to 2,5-dichloro-\n            <jats:italic>p<\/jats:italic>\n            -quinol and then to 5-chlorohydroxyquinol but converted 2,4,6-trichlorophenol only to 2,6-dichloro-\n            <jats:italic>p<\/jats:italic>\n            -quinol as the final product. TftD interacted with FADH\n            <jats:sub>2<\/jats:sub>\n            and retarded its rapid oxidation by O\n            <jats:sub>2<\/jats:sub>\n            . A spectrum of possible TftD-bound FAD-peroxide was identified, indicating that the peroxide is likely the active oxygen species attacking the aromatic substrates. The reclassification of the two enzymes further supports the new discovery of FADH\n            <jats:sub>2<\/jats:sub>\n            -utilizing enzymes, which have homologues in the domains\n            <jats:italic>Bacteria<\/jats:italic>\n            and\n            <jats:italic>Archaea<\/jats:italic>\n            .\n          <\/jats:p>","DOI":"10.1128\/jb.185.9.2786-2792.2003","type":"journal-article","created":{"date-parts":[[2003,4,16]],"date-time":"2003-04-16T21:03:42Z","timestamp":1050527022000},"page":"2786-2792","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":80,"title":["Characterization of Chlorophenol 4-Monooxygenase (TftD) and NADH:Flavin Adenine Dinucleotide Oxidoreductase (TftC) of\n            <i>Burkholderia cepacia<\/i>\n            AC1100"],"prefix":"10.1128","volume":"185","author":[{"given":"Michelle R.","family":"Gisi","sequence":"first","affiliation":[{"name":"School of Molecular Biosciences, Washington State University, Pullman, Washington"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Luying","family":"Xun","sequence":"additional","affiliation":[{"name":"School of Molecular Biosciences, Washington State University, Pullman, Washington"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"crossref","first-page":"739","DOI":"10.1111\/j.1432-1033.1997.t01-1-00739.x","volume":"249","year":"1997","unstructured":"Becker, D., T. 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