Conversion of NfsA, the major Escherichia coli nitroreductase, to a flavin reductase with an activity similar to that of Frp, a flavin reductase in Vibrio harveyi, by a single amino acid substitution.
NfsA is the major oxygen-insensitive nitroreductase of Escherichia coli, similar in amino acid sequence to Frp, a flavin reductase of Vibrio harveyi. Here, we show that a single amino acid substitution at position 99, which may destroy three hydrogen bonds in the putative active center, transforms NfsA from a nitroreductase into a flavin reductase that is as active as the authentic Frp and a tartrazine reductase that is 30-fold more active than wild-type NfsA.
Zenno
S
S
Department of Biophysics and Biochemistry, Graduate School of Science, University of Tokyo, Japan. tmichiue@hgc.ims.u-tokyo.ac.jp
Kobori
T
T
Tanokura
M
M
Saigo
K
K
D000595:Amino Acid Sequence; D004926:Escherichia coli / Q000201:enzymology*; D038181:FMN Reductase; D038702:Follistatin-Related Proteins; D006023:Glycoproteins / Q000737:chemistry / Q000378:metabolism*; D006860:Hydrogen Bonding; D007700:Kinetics; D008969:Molecular Sequence Data; D009247:NADH, NADPH Oxidoreductases / Q000096:biosynthesis* / Q000737:chemistry / Q000378:metabolism; D009601:Nitroreductases / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D014733:Vibrio / Q000201:enzymology*
D038702:Follistatin-Related Proteins; D006023:Glycoproteins; D038181:FMN Reductase; D009247:NADH, NADPH Oxidoreductases; D009601:Nitroreductases