NSF Award
2204024
With the support of the Chemistry of Life Processes program in the Division of Chemistry, Professors Brian Bennett and Adam Fiedler from Marquette University and Professor Richard Holz from the Colorado School of Mines will investigate the chemical function of the enzyme nitrile hydratase (NHase), which catalyzes the reaction of nitrile compounds with water (hydration) to give the corresponding amides. NHases hydrate a range of natural and synthetic nitriles with strict selectivity under mild conditions. As such, these enzymes have several industrial and pharmaceutical applications, including the removal of chemicals and pesticides from the environment. To expand the biocatalytic potential of NHases, a better understanding of the catalytic mechanism is essential. Professors Bennett, Fiedler and Holz will employ a combination of experimental and computational approaches to detect and probe intermediates generated during the hydration reaction. In addition, the assembly of the enzyme active site, which contains a cobalt or iron ion, will be examined. The project will provide exceptional training for undergraduate and graduate-level scientists. The emphasis on green chemistry and biophysical methods will serve as the basis for community engagement activities. The award will also enhance the research and educational infrastructures at both Marquette University and the Colorado School of Mines. <br/><br/>This research project seeks to gain molecular-level insights into NHase catalysis through a combination of kinetic, spectroscopic, biochemical, computational, and X-ray crystallographic methods. The studies will (i) test competing prevailing mechanistic hypotheses and refine the catalytic mechanism by interrogating observed intermediates, (ii) determine the catalytic function of proton transfer and the protonation states of ionizable groups in the active site, and (iii) elucidate steps in the maturation process of the metal-based (Fe or Co) active sites. The insights gained from the studies are expected to aid in the development of green catalytic approaches to hydrolytic reactions in nature and industry.<br/><br/>This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.
