Fast protein liquid chromatography system for protein purification

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All living things have specialized molecules, called enzymes, to carry out specific biological functions. A number of enzymes have evolved to use radicals (highly reactive lone electrons) to perform biological transformations. The advantage of radicals lies in their highly reactive nature, and therefore ability to carry out reactions that are chemically difficult and require a lot of energy. However, radicals react indiscriminately, which can damage the enzyme. Enzymes are dynamic systems; hence, the timing of internal motions must be exquisitely synchronized with chemical events to prevent the radical extinguishing itself. The goal of the research laboratory is to study how enzymes orchestrate radical chemistry with large structural rearrangements. To do this, the laboratory will study an enzyme that contains a vitamin B12-derivative, which it uses to initiate radical chemistry through breakage of an unusual carbon-cobalt bond. The radicals are then propagated to another vitamin-derived molecule (vitamin B6), nestled in the enzyme at a distant location. Fast-reaction spectroscopic techniques along with chemical synthesis of novel compounds similar to B12 will allow for an in-depth investigation into the kinetic and dynamic behaviour of the enzyme. From this research, models for radical-based catalysis will be derived that can be applied to other enzyme systems. The research will also enable us to re-engineer enzymes to carry out other chemical transformations that are applicable to drug synthesis or other industrial purposes. However, to study an individual enzyme, it first must be separated from all of the other thousands of proteins and enzymes in the bacterial cell. A high-pressure liquid chromatography system, like the ÄKTA FPLC is designed to do just that. The system is fully automated and is able to separate proteins at high resolution, quickly and reproducibly. The system is widely used in industry and academia, and HQP who gain expertise on the ÄKTA system will be better prepared for future careers in research and development. An ÄKTA FPLC, or a comparable system, is currently not available at UBC Okanagan.