title: Characterization of the staphylococcal bacteriophage lysin CHAP(K).
abstract: OBJECTIVE
  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.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Biological Sciences, Cork Institute of Technology, Cork, Ireland.
authors.forename: M
authors.initials: M
authors.lastname: Fenton
authors.forename: R P
authors.initials: RP
authors.lastname: Ross
authors.forename: O
authors.initials: O
authors.lastname: McAuliffe
authors.forename: J
authors.initials: J
authors.lastname: O'Mahony
authors.forename: A
authors.initials: A
authors.lastname: Coffey
mesh_terms: D000900:Anti-Bacterial Agents / Q000302:isolation & purification / Q000494:pharmacology*; D004798:Enzymes / Q000302:isolation & purification / Q000494:pharmacology*; D008248:Lysostaphin / Q000494:pharmacology; D008826:Microbial Sensitivity Tests; D013204:Staphylococcus Phages / Q000201:enzymology*; D013211:Staphylococcus aureus / Q000187:drug effects*
chemical_list: D000900:Anti-Bacterial Agents; D004798:Enzymes; C028806:lysin, streptococcus bacteriophage; D008248:Lysostaphin