Biochemical characterization of NfsA, the Escherichia coli major nitroreductase exhibiting a high amino acid sequence homology to Frp, a Vibrio harveyi flavin oxidoreductase

被引:170
作者
Zenno, S
Koike, H
Kumar, AN
Jayaraman, R
Tanokura, M
Saigo, K
机构
[1] UNIV TOKYO,GRAD SCH SCI,BIOTECHNOL RES CTR,BUNKYO KU,TOKYO 113,JAPAN
[2] CHISSO CORP,YOKOHAMA RES CTR,YOKOHAMA,KANAGAWA 236,JAPAN
[3] MADURAI KAMARAJ UNIV,SCH BIOL SCI,MADURAI 625021,TAMIL NADU,INDIA
关键词
D O I
10.1128/jb.178.15.4508-4514.1996
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
We identified the nfsA gene, encoding the major oxygen-insensitive nitroreductase in Escherichia coli, and determined its position on the E. coli map to be 19 min. We also purified its gene product, NfsA, to homogeneity. It was suggested that NfsA is a nonglobular protein with a molecular weight of 26,799 and is associated tightly with a flavin mononucleotide. Its amino acid sequence is highly similar to that of Frp, a flavin oxidoreductase from Vibrio harveyi (B. Lei, M. Liu, S. Huang, and S.-C. Tu, J. Bacteriol. 176:3552-3558, 1994), an observation supporting the notion that E. coli nitroreductase and luminescent-bacterium flavin reductase families are intimately related in evolution. Although no appreciable sequence similarity was detected between two E. coli nitroreductases, NfsA and NfsB, NfsA exhibited a low level of the flavin reductase activity and a broad electron acceptor specificity similar to those of NfsB. NfsA reduced nitrofurazone by a ping-pong Bi-Bi mechanism possibly to generate a two-electron transfer product.
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页码:4508 / 4514
页数:7
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