Purification and characterization of beta-lactamase from Neisseria gonorrhoeae from clinical samples
De Castillo, M.C.; Islas, M.I.; de Nader, O.M.; de Ruiz-Holgado, A.P.
Revista Latinoamericana de Microbiologia 43(2): 70-75
2001
ISSN/ISBN: 0187-4640 PMID: 17061490 Document Number: 536260
Beta-Lactamase was isolated from Neisseria gonorrhoeae, obtained from male patients with gonococcic urethritis. Biochemical properties of the enzyme were studied. The enzyme was purified 38-fold by ammonium sulphate precipitation and using Sephadex G75 and DEAE-cellulose columns. The purified extract exhibited a single band by polyacrylamide gel electrophoresis. Maximum enzyme activity was obtained at 37 degrees C and pH 7.0-7.2 in 50 mM phosphate buffer. Addition of Ni2+, Fe2+, Fe3+, Mn2+ and p-chloromercurybenzoate to the reaction buffer partially inhibited beta-lactamase activity, whereas Hg2+ and EDTA produced complete inhibition. The molecular weight was estimated to be 35,000 Da and the pI of the enzyme was 5.4.