Mechanisms of beta-lactamase inhibition and heterotropic allosteric regulation of an engineered beta-lactamase-MBP fusion protein

Expression of beta-lactamases is the most common mechanisms of beta-lactam antibiotic resistance clinically encountered. Both the numbers and types of beta-lactamases are growing rapidly. Therefore, there is an urgent need to develop more potent antibiotics and beta-lactamase inhibitors. My thesis work was focused on exploring the structural basis of beta-lactamases overcoming new beta-lactam antibiotics and inhibitors, on exploring this knowledge to develop improved inhibitors, and on exploring other ways of regulation (molecular switch). The SHV-1 beta-lactamase, KPC-2 carbapenemase and inhibitor-resistant S130G SHV will be used as the model system to characterize the active site chemistry of enzyme-inhibitor complexes. Three types of inhibitors were tested including the ones exploiting the function of R1 substituent, exploiting the function of R2 substituent, and exploiting transition state inhibitors. Also TEM-MBP fusion protein will be used to investigate the mechanisms of heterotropic allosteric regulation.