Open AccessDetection and purification of a catalase‐peroxidase from Mycobacterium sp. Pyr‐1
Author(s) -
Rafii Fatemeh,
Lunsford Pamala,
Hehman Gery,
Cerniglia Carl E
Publication year - 1999
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1999.tb13515.x
Subject(s) - catalase , peroxidase , chemistry , enzyme , sodium azide , biochemistry , isoniazid , chromatography , nuclear chemistry , medicine , tuberculosis , pathology
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 H 2 O 2 . 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 H 2 O 2 . 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°C.