Development of Advanced Chimeric Endolysin to Control Multidrug-Resistant through Domain Shuffling.

The increase in the prevalence of multidrug-resistant (MDR) with strong biofilm-forming capacity poses a serious public health concern. Endolysins derived from bacteriophages are a promising solution for antibiotic resistance problems. However, some natural staphylococcal endolysins have several shortcomings, such as low solubility and high sequence homology among domains. To overcome these limitations, we constructed a hybrid endolysin library by swapping an enzymatically active domain (EAD) and a cell wall binding domain (CBD) of 12 natural staphylococcal endolysins. We found a novel chimeric endolysin, ClyC, which showed enhanced lytic activity against compared to its parental endolysin forms. ClyC also exhibited strong antibacterial activity against in various biomatrices, such as milk and blood. Moreover, the treatment of chimeric endolysin effectively eradicated biofilms of multidrug-resistant bacteria, including methicillin-resistant (MRSA), (MRSE), and clinical isolates. In an in vivo mouse infection model, ClyC showed effective protection capability against methicillin-resistant (MRSA) without any toxic effects. Taken together, our data suggest that the chimeric endolysin ClyC can be considered a potential antibacterial agent against multidrug-resistant and may have clinical relevance.