Development of the next generatation Artilysins, enzyme-based antibiotics against multidrug-resistant bacteria

This project will boost the development of Artilysins, novel enzyme-based antibiotics that respond to the unmet need for novel antibiotics against Gram-negative bacteria. An elegant biotechnological methodology (Golden Gate shuffling) will be integrated to accelerate the creation and analysis of novel candidate Artilysins, and this on a much larger scale than before. After its integration, we will be able to select the most optimized Artilysins for most diverse bacterial infections. Artilysins combine the power of endolysins and specific peptides. Endolysins are enzymes encoded by bacterial viruses (the enemy of our enemy) and kill the infected bacteria by peptidoglycan degradation. We fuse endolysins to selective peptides that transfer the endolysin to its substrate, the peptidoglycan layer, which is subsequently degraded. Artilysins kill bacteria with high efficiency, including multidrug-resistant strains and persisters, and are refractory to resistance development. The first generations of Artilysins have been constructed by lengthy, traditional cloning. Although successful in its finality, integration of the Golden Gate shuffling method will permit to construct and analyze several hundreds to thousands Artilysin in a single effort. A repository of Artilysins building blocks will be constructed and used for both precisely designed and randomly created Artilysins, together with the necessary tools for analysis on a large scale.