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KMID : 0545119930030030174
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Journal of Microbiology and Biotechnology
1993 Volume.3 No. 3 p.174 ~ p.180
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Production and Characterization of Extracellular β-Lactamase from Streptomyces aureofaciens SMF14
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Kim. Myung Kuk
Lee Kye-Joon
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Abstract
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A strain SMF14 producing an extracellular β-lactamase was isolated from soil and identified to be a strain of Streptomyces aureofaciens. β-lactamase was purified from the cell free culture broth through batchwise hydroxyapatite adsorption, anion exchange chromatography on DEAE Sephadex A-50, gel filtration on Sephadex G-75, and adsorption chromatography on hydroxyapatite. The molecular mass was estimated to be about 43 kDa by SDS-PAGE. The β-lactamase had substrate specificity to penicillins and it was inhibited by clavulanic acid, being classified to the group 2a of penicillinase. The optimal reaction pH and temperature were pH 6.0∼7.5 and 50℃. The K_m and V_max values of β-lactamase for penicillin G were calculated to be 1.72mM and 5.4× 10 exp (5)μM·min^-1, respectively.
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