Antimicrobial Activity of LysX and LysP Endolysins Against Pseudomonas syringae pv. syringae and Xanthomonas arboricola pv. juglandis

<i>Pseudomonas syringae</i> pv. <i>syringae</i> and <i>Xanthomonas arboricola</i> pv. <i>juglandis</i> are the causal agents of bacterial canker in cherry and walnut blight, respectively, which cause significant production losses worldwide. These diseases have traditionally been controlled by copper-based agrochemicals and, more recently, antibiotics. However, the prolonged use of these compounds has led to the emergence of resistant bacterial strains. The search for new, efficient, and environmentally friendly biocontrol alternatives has intensified. Phages are promising candidates due to their ability to specifically infect and lyse bacterial pathogens. Endolysin enzymes are responsible for bacterial cell wall degradation, and although they have been extensively studied in medical and veterinary contexts, their application in agriculture remains limited. In this study, 17 putative endolysins were identified from bacteriophages infecting <i>X. arboricola</i> pv. <i>juglandis</i> and <i>P. syringae</i> pv. <i>syringae</i>. Based on conserved domain analyses, 12 were classified as glycosidases, four as amidases, and one as an endopeptidase. From these, a recombinant amidase (LysP) and a recombinant glycosidase (LysX) were expressed in <i>E. coli</i>, purified, and evaluated as pure enzymes. Both endolysins exhibited significant antimicrobial activity, reducing <i>P. syringae</i> pv. <i>syringae</i> viability by 62-78.3% and <i>X. arboricola</i> pv. <i>juglandis</i> viability by 51.5-53.1%, respectively. These findings highlight these recombinant endolysins as promising candidates for the development of biocontrol strategies against bacterial plant pathogens.