'Artilysation' of endolysin λSa2lys strongly improves its enzymatic and antibacterial activity against streptococci

Sci Rep. 2016 Oct 24;6:35382. doi: 10.1038/srep35382.

Abstract

Endolysins constitute a promising class of antibacterials against Gram-positive bacteria. Recently, endolysins have been engineered with selected peptides to obtain a new generation of lytic proteins, Artilysins, with specific activity against Gram-negative bacteria. Here, we demonstrate that artilysation can also be used to enhance the antibacterial activity of endolysins against Gram-positive bacteria and to reduce the dependence on external conditions. Art-240, a chimeric protein of the anti-streptococcal endolysin λSa2lys and the polycationic peptide PCNP, shows a similar species specificity as the parental endolysin, but the bactericidal activity against streptococci increases and is less affected by elevated NaCl concentrations and pH variations. Time-kill experiments and time-lapse microscopy demonstrate that the killing rate of Art-240 is approximately two-fold higher compared to wildtype endolysin λSa2lys, with a reduction in viable bacteria of 3 log units after 10 min. In addition, lower doses of Art-240 are required to achieve the same bactericidal effect.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anti-Bacterial Agents / chemistry*
  • Anti-Bacterial Agents / pharmacology
  • Bacteriophages / enzymology
  • Circular Dichroism
  • Endopeptidases / chemistry*
  • Endopeptidases / pharmacology
  • Gram-Negative Bacteria / drug effects
  • Gram-Positive Bacteria / drug effects
  • Hydrogen-Ion Concentration
  • Microbial Sensitivity Tests
  • Peptidoglycan / chemistry
  • Recombinant Proteins / chemistry
  • Salts / chemistry
  • Streptococcus / drug effects*
  • Temperature

Substances

  • Anti-Bacterial Agents
  • Peptidoglycan
  • Recombinant Proteins
  • Salts
  • Endopeptidases
  • endolysin