Characterization of MnpC, a hydroquinone dioxygenase likely involved in the meta-nitrophenol degradation by Cupriavidus necator JMP134.
Cupriavidus necator JMP134 utilizes meta-nitrophenol (MNP) as the sole source of carbon, nitrogen, and energy. The metabolic reconstruction of MNP degradation performed in silico suggested that MnpC might have played an important role in MNP degradation. In order to experimentally confirm the prediction, we have now characterized the mnpC-encoded (amino)hydroquinone dioxygenase involved in the ring-cleavage reaction of MNP degradation. Real-time PCR analysis indicated that mnpC played an essential role in MNP degradation. MnpC was purified to homogeneity as an N-terminal six-His-tagged fusion protein, and it was proved to be a dimer as demonstrated by gel filtration. MnpC was a Fe(2+)- and Mn(2+)-dependent dioxygenase, catalyzing the ring-cleavage of hydroquinone to 4-hydroxymuconic semialdehyde in vitro and proposed as an aminohydroquinone dioxygenase involved in MNP degradation in vivo. Phylogenetic analysis suggested that MnpC diverged from the other (chloro)hydroquinone dioxygenases at an earlier point, which might result in the preference for its physiological substrate.
Yin
Ying
Y
State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, China.
Zhou
Ning-Yi
NY
D000595:Amino Acid Sequence; D002244:Carbon; D003001:Cloning, Molecular; D020118:Cupriavidus necator / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism*; D049308:Dioxygenases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D005798:Genes, Bacterial; D006873:Hydroquinones / Q000378:metabolism*; D009584:Nitrogen; D009596:Nitrophenols / Q000378:metabolism*; D009601:Nitroreductases / Q000235:genetics / Q000378:metabolism; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013379:Substrate Specificity
D006873:Hydroquinones; D009596:Nitrophenols; D002244:Carbon; D049308:Dioxygenases; D009601:Nitroreductases; D009584:Nitrogen; C049017:3-nitrophenol; C031927:hydroquinone
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