title: Detection and purification of a catalase-peroxidase from Mycobacterium sp. Pyr-1.
abstract: A catalase-peroxidase from Mycobacterium sp. Pyr-1, a strain capable of growth on pyrene, was purified to homogeneity by anion exchange and hydroxyapatite column chromatography. The enzyme, like the M. tuberculosis T-catalase, reduced nitroblue tetrazolium in the presence of isoniazid (INH) and H2O2. It also oxidized 3,3',5,5'-tetramethylbenzidine and other substrates of the catalase-peroxidase of M. tuberculosis in the presence of either tert-butyl hydroperoxide or H2O2. It had a UV/ visible absorption spectrum (Soret peak at 406 nm) similar to that of the catalase-peroxidase of M. tuberculosis (Soret peak at 408 nm) and identical to that of the catalase-peroxidase of M. smegmatis. After electrophoresis on non-denaturing gels the enzyme showed one single protein band with both catalase and peroxidase activity, which were lost after electrophoresis on SDS-PAGE. The enzyme was inhibited by sodium azide, glutathione, 2-mercaptoethanol, and isoniazid, but not by isonicotinic acid. The optimum enzyme activity was obtained at pH 4.5 and at 25 degrees C.
authors.affiliation: Division of Microbiology, National Center for Toxicological Research, FDA, Jefferson, AR 72079, USA. frafii@nctr.fda.gov
authors.forename: F
authors.initials: F
authors.lastname: Rafii
authors.forename: P
authors.initials: P
authors.lastname: Lunsford
authors.forename: G
authors.initials: G
authors.lastname: Hehman
authors.forename: C E
authors.initials: CE
authors.lastname: Cerniglia
mesh_terms: D000995:Antitubercular Agents / Q000494:pharmacology; D002374:Catalase / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism; D004352:Drug Resistance, Microbial; D006863:Hydrogen-Ion Concentration; D007538:Isoniazid / Q000494:pharmacology; D009161:Mycobacterium / Q000187:drug effects / Q000201:enzymology* / Q000254:growth & development; D009195:Peroxidase / Q000302:isolation & purification* / Q000378:metabolism; D013696:Temperature
chemical_list: D000995:Antitubercular Agents; D002374:Catalase; D009195:Peroxidase; D007538:Isoniazid