This CAREER award supports research by Professor Xiaotang Wang of Florida International University to determine the structural basis for chloroperoxidase-catalyzed enantioselective transformations. Assignments for all hyperfine-shifted proton resonances of the heme and heme-pocket amino acids of chloroperoxidase (CPO) will be made by a combination of two-dimensional NOESY and COSY techniques to determine connectivities. Highly deuterated CPO samples will be purified to facilitate resonance assignments. Three and four dimensional NMR experiments will also be attempted with 15N- and/or 13C-labeled CPO samples to help assign resonances and develop a structural characterization of CPO in solution. Detailed structural characterization of the CPO-substrate complexes will then be carried out through enhanced nuclear relaxation, internuclear NOE, saturation-transferred NOE, heteronuclear single or multiple quantum correlation, optical, and fluorescence spectroscopic methods. These results will be combined to develop a clear picture of the interactions between CPO and its various organic substrates, thereby providing the structural basis for the CPO-catalyzed chiral transformations. These results will be used in the design of mutants that can be used as new and cost-effective enzymes for making important chiral synthons.Professor Wang's education plan introduces modern research techniques and innovative evaluation methods into courses in order to keep students abreast of the current knowledge of chemistry and enhance their understanding of chemical principles. New course material (Computers in Chemistry Laboratory Instruction, CCLI) will be adopted for the general chemistry laboratory course. In addition, a new graduate course, special topics in inorganic chemistry (bioinorganic chemistry), will be developed. A summer research program for high school students will also be undertaken. Understanding how chloroperoxidase generates chiral transformation could enable biotechnological applications of metalloenzymes, especially in the design of new enzyme catalysts for synthesizing chiral materials of industrial and medicinal importance. The education plan will develop both research and new courses in bioinorganic chemistry at this minority-serving institution.
