Journal
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
Volume 67, Issue 8, Pages 2245-2254Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.jafc.9b00173
Keywords
biodegradation; dechlorination; Cupriavidus nantongensis; tcpA; fre
Funding
- National Natural Science Foundation of China [31471792]
- USDA Hatch project [WAW05020-H]
- Graduate Innovation Fund of Anhui Agricultural University [2018yjs-54]
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Chlorpyrifos is one of the most used organophosphorus insecticides. It is commonly degraded to 3,5,6-trichloro-2-pyridinol (TCP), which is water-soluble and toxic. Bacteria can degrade chlorpyrifos and TCP, but the biodegradation mechanism has not been well-characterized. Recently isolated Cupriavidus nantongensis X1(T) can completely degrade 100 mg/L chlorpyrifos and 20 mg/L TCP with half-lives of 6 and 8 h, respectively. We annotated a complete gene cluster responsible for TCP degradation in recently sequenced strain X1(T). Two key genes, tcpA and fre, were cloned from X1(T) and transferred and expressed in Escherichia coil BL21(DE3). Degradation of TCP by X1(T) whole cell was compared with that by the enzymes 2,4,6-trichlorophenol monooxygenase and NAD(P)H:flavin reductase expressed and purified from E. coli BL21(DE3). Novel metabolites of TCP were isolated and characterized, indicating stepwise dechlorination of TCP, which was confirmed by TCP disappearance, mass balance, and detection and formation kinetics of chloride ion from TCP.
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