title: Purification and characterization of an enzyme from Mycobacterium sp. Pyr-1, with nitroreductase activity and an N-terminal sequence similar to lipoamide dehydrogenase.
abstract: Mycobacterium sp. Pyr-1 produces an enzyme with nitroreductase activity that reduces 1-nitropyrene and 4-nitrobenzoic acid to the corresponding aromatic amines. This enzyme was constitutive and required NADH; and its activity was enhanced by FAD. It was inhibited by antimycin A, dicumarol, and o-iodosobenzoic acid; and it was inactivated by ammonium sulfate precipitation. After purification to homogeneity, the protein produced a single band on native and SDS-polyacrylamide gels and had a single amino-terminal sequence. The N-terminal amino acid sequence was identical to the corresponding sequences of the lipoamide dehydrogenases of M. leprae, M. tuberculosis and Corynebacterium glutamicum. The amino-terminal sequence was also similar to lipoamide dehydrogenases from M. smegmatis and several other bacteria. The amino acid sequence of an internal peptide (12 of 13 amino acids) was nearly identical to the corresponding sequences of lipoamide dehydrogenases from M. leprae and M. tuberculosis and was similar to those of C. glutamicum, Streptomyces coelicolor and S. seoulensis. The data show that a unique lipoamide dehydrogenase in Mycobacterium sp. Pyr-1, which differs from classic (Type I) bacterial nitroreductases, reduces aromatic nitro compounds to aromatic amines.
authors.affiliation: Division of Microbiology, National Center for Toxicological Research, US Food and Drug Administration, Jefferson AR 72079, USA. frafii@nctr.fda.gov
authors.forename: F
authors.initials: F
authors.lastname: Rafii
authors.forename: G
authors.initials: G
authors.lastname: Hehman
authors.forename: P
authors.initials: P
authors.lastname: Lunsford
mesh_terms: D000595:Amino Acid Sequence; D008058:Dihydrolipoamide Dehydrogenase / Q000737:chemistry* / Q000378:metabolism; D008969:Molecular Sequence Data; D009161:Mycobacterium / Q000201:enzymology* / Q000254:growth & development; D009601:Nitroreductases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*
chemical_list: D009601:Nitroreductases; D008058:Dihydrolipoamide Dehydrogenase