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


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.


  1. Archer, G. L., G. J. Vazquez and J. L. Johnston. 1980. Antibiotic prophylaxis of experimental endocarditis due to methicillin-resistant Staphylococcus epidermidis. J. Infect. Dis. 142, 725-731
  2. Baddour, L. M., T. N. Phillips and A. L. Bisno, 1986. Coagulase-negative staphylococcal endocarditis. Occurrence in patients with mitral valve prolapse. Arch. Intern. Med. 146, 119-121
  3. Benini, S., W. R. Rypniewski, K. S. Wilson, S. Miletti, S. Ciurli and S. Mangani. 1999. A new proposal for urease mechanism based on the crystal structures of the native and inhibited enzyme from Bacillus pasteurii: why urea hydrolysis costs two nickels. Structure. 7, 205-216
  4. Dunne, W. M., Jr. 2002. Bacterial adhesion: seen any good biofilms lately? Clin. Microbiol. Rev. 15, 155-166
  5. Gatermann, S., J. John and R. Marre. 1989. Staphylococcus saprophyticus urease: characterization and contribution to uropathogenicity in unobstructed urinary tract infection of rats. Infect. Immun. 57, 110-116
  6. Gatermann, S. and R. Marre. 1989. Cloning and expression of Staphylococcus saprophyticus urease gene sequences in Staphylococcus carnosus and contribution of the enzyme to virulence. Infect. Immun. 57, 2998-3002
  7. Gill, S. R., D. E. Fouts, G. L. Archer, E. F. Mongodin, R. T. Deboy, J. Ravel, I. T. Paulsen, J. F. Kolonay, L. Brinkac, M. Beanan, R. J. Dodson, S. C. Daugherty, R. Madupu, S. V. Angiuoli, A. S. Durkin, D. H. Haft, J. Vamathevan, H. Khouri, T. Utterback, C. Lee, G. Dimitrov, L. Jiang, H. Qin, J. Weidman, K. Iran, K. Kang, I. R. Hance, K. E. Nelson and C. M. Fraser. 2005. Insights on evolution of virulence and resistance from the complete genome analysis of an early methicillin-resistant Staphylococcus aureus strain and a biofilm-producing methicillin-resistant Staphylococcus epidermidis strain. J. Bacteriol. 187, 2426-2438
  8. Goldmann, D. A. and G. B. Pier. 1993. Pathogenesis of infections related to intravascular catheterization. Clin. Microbiol. Rev. 6, 176-192
  9. Gruter, L., M. Endres and S. Gatermann. 1992. Cloning and expression of Staphylococcus epidermidis urease gene sequences in Staphylococcus carnosus. FEMS Microbiol. Lett. 72, 33-35
  10. Huebner, J. and D. A. Goldmann. 1999. Coagulase-negative staphylococci: role as pathogens. Annu. Rev. Med. 50, 223-236
  11. Jabri, E., M. B. Carr, R. P. Hausinger and P. A. Karplus. 1995. The crystal structure of urease from Klebsiella aerogenes. Science 268, 998-1004
  12. Kloos, W. E. and T. L. Bannerman. 1994. Update on clinical significance of coagulase-negative staphylococci. Clin. Microbiol. Rev. 7, 117-140
  13. Laemmli, U. K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680-685
  14. Leighton, P. M. and J. A. Little. 1986. Identification of coagulase-negative Staphylococci isolated from urinary tract infections. Am. J. Clin. Pathol. 85, 92-95
  15. Lowry, O. H., N. J. Rosebrough, A. L. Farr and R. J. Randall. 1951. Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193, 265-275
  16. Martin, M. A., M. A. Pfaller and R. P. Wenzel. 1989. Coagulase-negative staphylococcal bacteremia. Mortality and hospital stay. Ann. Intern. Med. 110, 9-16
  17. Mobley, H. L. and R. P. Hausinger. 1989. Microbial ureases: significance, regulation, and molecular characterization. Microbiol. Rev. 53, 85-108
  18. Mobley, H. L., M. D. Island and R. P. Hausinger. 1995. Molecular biology of microbial ureases. Microbiol. Rev. 59, 451-480
  19. O'Gara, J. P. and H. Humphreys. 2001. Staphylococcus epidermidis biofilms: importance and implications. J. Med. Microbiol. 50, 582-587
  20. Rubin, J., W. A. Rogers, H. M. Taylor, E. D. Everett, B. F. Prowant, L. V. Fruto and K. D. Nolph. 1980. Peritonitis during continuous ambulatory peritoneal dialysis. Ann. Intern. Med. 92, 7-13
  21. Schafer, U. K. and H. Kaltwasser. 1994. Urease from Staphylococcus saprophyticus: purification, characterization and comparison to Staphylococcus xylosus urease. Arch. Microbiol. 161, 393-399
  22. Schierholz, J. M. and J. Beuth. 2001. Implant infections: a haven for opportunistic bacteria. J. Hosp. Infect. 49, 87-93
  23. Tacconelli, E., M. Tumbarello, M. Pittiruti, F. Leone, M. B. Lucia, R. Cauda and L. Ortona. 1997. Central venous catheter-related sepsis in a cohort of 366 hospitalised patients. Eur. J. Clin. Microbiol. Infect. Dis. 16, 203-209
  24. Vuong, C. and M. Otto. 2002. Staphylococcus epidermidis infections. Microbes. Infect. 4, 481-489
  25. Weatherburn, M. W. 1967. Phenol-hypochlorite reaction for determination of ammonia. Anal. Chem. 39, 971-974
  26. Zhang, Y. Q., S. X. Ren, H. L. Li, Y. X. Wang, G. Fu, J. Yang, Z. Q. Qin, Y. G. Miao, W. Y. Wang, R. S. Chen, Y. Shen, Z. Chen, Z. H. Yuan, G. P. Zhao, D. Qu, A. Danchin and Y. M. Wen. 2003. Genome-based analysis of virulence genes in a non-biofilm-forming Staphylococcus epidermidis strain (ATCC 12228). Mol. Microbiol. 49, 1577-1593

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