Analysis of outer membrane permeability of Pseudomonas aeruginosa and bactericidal activity of endolysins KZ144 and EL188 under high hydrostatic pressure.
The parameters influencing outer membrane permeability of Pseudomonas aeruginosa PAO1 under high hydrostatic pressure were quantified and optimized, using fusion between a specific A1gamma peptidoglycan-binding domain and green fluorescent protein (PBD-GFP). Based on the obtained data, optimal conditions were defined to assess the synergistic bactericidal action between high hydrostatic pressure and peptidoglycan hydrolysis by bacteriophage-encoded endolysins KZ144 and EL188. Under high hydrostatic pressure, both endolysins show similar inactivation of P. aeruginosa as the commonly used hen egg white lysozyme or slightly higher inactivation in the case of EL188 at 150 and 200 MPa. The partial contribution of pressure to the bacterial inactivation increases with higher pressure, while the partial contribution of the enzymes is maximal at the onset pressure of outer membrane permeabilization for the PBD-GFP protein (175 MPa). This study's results demonstrate the usefulness of this approach to determine optimal synergy for hurdle technology applications.
Briers
Yves
Y
Department of Biosystems, Division of Gene Technology, Katholieke Universiteit Leuven, Leuven, Belgium. yves.briers@biw.kuleuven.be
Cornelissen
Anneleen
A
Aertsen
Abram
A
Hertveldt
Kirsten
K
Michiels
Chris W
CW
Volckaert
Guido
G
Lavigne
Rob
R
D000900:Anti-Bacterial Agents / Q000302:isolation & purification / Q000494:pharmacology*; D001431:Bacteriological Techniques / Q000379:methods*; D001435:Bacteriophages / Q000201:enzymology; D002463:Cell Membrane Permeability / Q000187:drug effects*; D015169:Colony Count, Microbial; D010450:Endopeptidases / Q000302:isolation & purification / Q000494:pharmacology*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006874:Hydrostatic Pressure; D008856:Microscopy, Fluorescence; D009113:Muramidase / Q000494:pharmacology; D010457:Peptidoglycan / Q000378:metabolism; D011550:Pseudomonas aeruginosa / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000494:pharmacology
D000900:Anti-Bacterial Agents; D010457:Peptidoglycan; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; C421864:hen egg lysozyme; D009113:Muramidase; D010450:Endopeptidases; C001143:endolysin