Investigations of proteins important for antibiotic resistance

The objective of my first project was to investigate the interaction of a class D beta-lactamase with different Beta-lactams to explore if this family of enzymes is diffusion-limited or not. This project required isolation and purification of the OXA-10 beta-lactamase overexpressed in E. coli BL21 and the determination of kcat, Km, and Vmax parameters for different beta-lactam antibiotics at different viscosities of the reaction buffer. The other project that I worked on required the cloning of blaI gene into an expression vector, overexpression of BlaI in E. coli BL21, purification of this protein. This protein is a gene repressor, believed to be involved in inducible expression of beta-lactamases in S. aureus.