Characterization of the staphylococcal bacteriophage lysin CHAP(K).

Journal of applied microbiology (2011-08-05)
M Fenton, R P Ross, O McAuliffe, J O'Mahony, A Coffey
RESUMO

To develop an efficient purification strategy for the bacteriophage lysin CHAP(K) . To evaluate its antibacterial spectrum(,) enzymatic properties, optimal reaction conditions and lytic activity against live Staphlyococcus aureus. Recombinant CHAP(K) was purified to homogeneity by cation exchange chromatography, with yields of up to 10 mg from 1 l of Escherichia coli culture. The lytic spectrum of CHAP(K) includes all staphylococcal species and also members of the genera Micrcococcus, Streptococcus, Nesterenkonia, Arthrobacter, Leuconostoc and Carnobacterium. The enzyme was active from pH 6 to 11 with an optimum activity at pH 9, from 5 to 40°C, with an optimum activity at 15°C. When cell lysis by CHAP(K) and lysostaphin was compared over a concentration range of 2·5-10 μg ml⁻¹ using live Staph. aureus for 5 min at 37°C, CHAP(K) gave rise to greater turbidity reduction indicating that it works more rapidly than lysostaphin. This study describes in detail the purification and characteristics of the novel phage-derived enzyme CHAP(K) demonstrating that it has excellent biochemical properties as an anti-staphylococcal agent. Currently, there is a need for new antimicrobial agents due to the increasing worldwide prevalence of antibiotic resistance. Our findings demonstrate the potential for development of CHAP(K) as an alternative therapeutic against pathogenic staphylococci including MRSA.

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Sigma-Aldrich
Lysostaphin from Staphylococcus staphylolyticus, lyophilized powder, Protein 50-70 % by biuret, ≥500 units/mg protein
Sigma-Aldrich
Lysostaphin from Staphylococcus simulans, recombinant, expressed in E. coli, lyophilized powder
Sigma-Aldrich
Lysostaphin from Staphylococcus staphylolyticus, BioUltra, ≥97% (SDS-PAGE), Protein 40-60 % by biuret, ≥2,000 units/mg protein
Sigma-Aldrich
Lysostaphin from Staphylococcus staphylolyticus, aseptically filled