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Characterization of the staphylococcal bacteriophage lysin CHAPK

Author:
Fenton, M., Ross, R.P., McAuliffe, O., O’Mahony, J., Coffey, A.
Source:
Journal of applied microbiology 2011 v.111 no.4 pp. 1025-1035
ISSN:
1364-5072
Subject:
Staphylococcus aureus, antibacterial properties, antibiotic resistance, cation exchange chromatography, recombinant proteins, turbidity, Nesterenkonia, Carnobacterium, animal pathogenic bacteria, bacteriophages, Leuconostoc, endolysin, enzymatic reactions, pH, Streptococcus, anti-infective agents, Arthrobacter, purification methods, Escherichia coli
Abstract:
Aims: To develop an efficient purification strategy for the bacteriophage lysin CHAPK. To evaluate its antibacterial spectrum, enzymatic properties, optimal reaction conditions and lytic activity against live Staphlyococcus aureus. Methods and Results: Recombinant CHAPK 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 CHAPK 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 CHAPK 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, CHAPK gave rise to greater turbidity reduction indicating that it works more rapidly than lysostaphin. Conclusions: This study describes in detail the purification and characteristics of the novel phage‐derived enzyme CHAPK demonstrating that it has excellent biochemical properties as an anti‐staphylococcal agent. Significance and Impact of the Study: 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 CHAPK as an alternative therapeutic against pathogenic staphylococci including MRSA.
Agid:
70590