Purification and characterization of NfrA1, a Bacillus subtilis nitro/flavin reductase capable of interacting with the bacterial luciferase.
ipa-43d is a hypothetical gene identified by the Bacillus subtilis genome project (Mol. Microbiol. 10, 371-384 1993; Nature 390, 249-256 1997). The ipa-43d protein overexpressed in E. coli was purified to homogeneity and its properties were analyzed biochemically. The ipa-43d protein was found to be tightly associated with FMN and to be capable of reducing both nitrofurazone and FMN effectively. Although the ipa-43d protein catalysis obeys the ping-pong Bi-Bi mechanism, catalysis mode was changed to the sequential mechanism upon coupling with the bioluminescent reaction. Database search showed that B. subtilis possessed four genes (ipa-44d, ytmO, yddN, and yvbT), encoding proteins similar in amino acid sequence to LuxA and LuxB of Photobacterium fischeri, and, in particular, ipa-44d is immediately adjacent to the ipa-43d gene on the chromosome.
Zenno
S
S
Department of Biophysics and Biochemistry, Graduate School of Science, University of Tokyo, Japan.
Kobori
T
T
Tanokura
M
M
Saigo
K
K
D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000201:enzymology*; D001426:Bacterial Proteins; D005486:Flavin Mononucleotide / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D008156:Luciferases / Q000378:metabolism*; D008969:Molecular Sequence Data; D009583:Nitrofurazone / Q000378:metabolism; D009601:Nitroreductases / Q000302:isolation & purification* / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
D001426:Bacterial Proteins; D005486:Flavin Mononucleotide; D008156:Luciferases; D009601:Nitroreductases; C116226:NfrA1 protein, Bacillus subtilis; D009583:Nitrofurazone