Nitroreductase activity of ferredoxin reductase BphA4 from Dyella ginsengisoli LA-4 by catalytic and structural properties analysis.
Ferredoxin reductase BphA4 was well known as a component of biphenyl dioxygenase. However, there was little information about whether it could utilize nonphysiological oxidants as electron acceptors. In the present study, we reported the novel nitroreductase activity of BphA4(LA)₋₄. The homology model of ferredoxin reductase BphA4 from Dyella ginsengisoli LA-4 was constructed. According to the alignment of three-dimensional structures, it was supposed that BphA4(LA)₋₄ could function as nitroreductase. Recombinant His-tagged BphA4(LA)₋₄ was purified with a molecular mass of 49.6 ± 1 kDa. Biochemical characterization of purified BphA4(LA)₋₄ possessed the nitroreductase activity with the optimal temperature 50°C and pH 8.0. The substrate spectrum and kinetics indicated BphA4(LA)₋₄ could reduce several nitroaromatics with different apparent K(m) values: m-dinitrobenzene (560 μM), o-dinitrobenzene (1,060 μM), o-nitroaniline (1,570 μM), m-nitrobenzoic acid (1,300 μM) and m-nitrophenol (67 μM). The nitroreductase activity was further explained by docking studies, which was indicated that Arg 288 should play an important role in binding nitroaromatics. Moreover, there existed a good linear correlation between lnK(m) and calculated binding energy.
Qu
Yuanyuan
Y
Dalian University of Technology, China.
Zhou
Hao
H
Li
Ang
A
Ma
Fang
F
Zhou
Jiti
J
D050900:Ferredoxin-Nitrite Reductase / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008958:Models, Molecular; D008970:Molecular Weight; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D013696:Temperature; D044166:Xanthomonadaceae / Q000201:enzymology*
D011994:Recombinant Proteins; D050900:Ferredoxin-Nitrite Reductase