Characterization of Endolysin LyJH307 with Antimicrobial Activity against Streptococcus bovis

SIMPLE SUMMARY: Development of endolysin is a promising strategy because of having the ability to control problematic bacteria specifically. In this study, we developed and characterized the endolysin having lytic activity against Streptococcus bovis (S. bovis), which is one of the initiators of ruminal acidosis. Based on our findings, endolysin LyJH307 showed potent lytic activity in ruminal pH range and ruminal temperature. In addition, LyJH307 was effective against not only S. bovis isolated from rumen, but also several S. bovis groups. We suggest that LyJH307 may have a lytic effect in the ruminal condition and prevent acute ruminal acidosis by controlling S. bovis specifically. ABSTRACT: Streptococcus bovis (S. bovis) is one of the critical initiators of acute acidosis in ruminants. Therefore, we aimed to develop and characterize the endolysin LyJH307, which can lyse ruminal S. bovis. We tested the bactericidal activity of recombinant LyJH307 against S. bovis JB1 under a range of pH, temperature, NaCl, and metal ion concentrations. In silico analyses showed that LyJH307 has a modular design with a distinct, enzymatically active domain of the NLPC/P60 superfamily at the N-terminal and a cell wall binding domain of the Zoocin A target recognition domain (Zoocin A_TRD) superfamily at the C-terminal. The lytic activity of LyJH307 against S. bovis JB1 was the highest at pH 5.5, and relatively higher under acidic, than under alkaline conditions. LyJH307 activity was also the highest at 39 °C, but was maintained between 25°C and 55°C. LyJH307 bactericidal action was retained under 0-500 mM NaCl. While the activity of LyJH307 significantly decreased on treatment with ethylenediaminetetraacetic acid (EDTA), it was only restored with supplementation of 10 mM Ca(2+). Analyses of antimicrobial spectra showed that LyJH307 lysed Lancefield groups D (S. bovis group and Enterococcus faecalis) and H (S. sanguinis) bacteria. Thus, LyJH307 might help to prevent acute ruminal acidosis.