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Revista Latinoamericana de Microbiología

2001, Number 2

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Microbiología 2001; 43 (2)

Purificación y caracterización de la Β-lactamasa de Neisseria gonorrhoeae proveniente de muestras clínicas

Castillo MC, Islas MI, Nader OM, Ruiz-Holgado AP

Language: English
References: 33
Page: 70-75
PDF size: 135.43 Kb.


ABSTRACT

Β-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°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 Β-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.


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