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Purification and Characterization of the Staphylococcus epidermidis Urease

Staphylococcus epidermidis urease의 정제 및 생화학적 특성에 관한 연구

  • 민선희 (대구가톨릭대학교 약학대학) ;
  • 이만형 (대구가톨릭대학교 약학대학)
  • Published : 2007.04.25

Abstract

Staphylococcus epidermidis is a coagulase-negative, gram-positive bacterium that normally inhabits the human skin. S. epidermidis is also known to be an opportunistic pathogen in infections of various indwelling medical devices. This report describes purification and characterization of the urease of S. epidermidis urease, which may act as a virulence factor. The urease from S. epidermidis was purified 1,127 fold by using DEAE-Sepharose, Phenyl-Sepharose, Mono-Q and Superdex HR200 column chromatography. The specific activity of the purified enzyme was 993.8 U/mg. Michaelis constant($K_m$) of the enzyme was estimated to be 8.5 mM urea by using Lineweaver-Burke double reciprocal plot. The native molecular weight of the urease was shown to be 255 kD by using Superose 6HR gel filtration chromatography and the purified enzyme contained 2.2 nickel ions per catalytic unit. The overall stoichiometry of the enzyme subunits appears to be $(\alpha\beta\gamma)_3$, which is consistent with the enzymes from other bacteria sources.

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