Improving Alkene Reductases for Applications in Asymmetric Synthesis

通过这一奖项，化学合成计划支持佛罗里达大学化学系的乔恩·D·斯图尔特教授开发用于不对称共轭烯烃还原的新型酶催化剂。到目前为止，这些酶对合成化学的总体影响并不大，因为野生型酶只接受有限范围的底物，提供一种可能的立体异构体，并且缺乏长期稳定性。为了克服这些问题，现有的黄素依赖的还原酶将在最有可能产生改进突变体的氨基酸位置上进行系统的变化。X射线结晶学将被用来了解底物如何结合在改良酶的活性部位。更广泛的影响包括指导高中生(UF大学前培训计划)、本科生(NSF REU)、研究生和博士后研究人员，包括化学领域历史上代表性不足的群体的成员。此外，这项工作还将为这些同事提供化学、X射线结晶学、分子生物学和生物化学方面的多样化培训。就更广泛的科学影响而言，这项工作旨在寻找高效、对环境无害的方法来生产用于医药、农用化学品和工业化学品应用的手性构建块。此外，结合蛋白质工程、结构研究和催化分析所获得的见解将提供有关黄素蛋白的新信息，黄素蛋白是一类重要的生物催化剂，参与了化学/生物界面上的一系列不同反应。

With this award, the Chemical Synthesis program is supporting Professor Jon D. Stewart of the Department of Chemistry at the University of Florida to develop new enzyme catalysts for asymmetric conjugated alkene reductions. To date, the overall impact of these enzymes on synthetic chemistry has been modest because the wild-type enzymes accept only a limited range of substrates, provide one possible stereoisomer and lack long-term stability. To overcome these problems, existing flavin-dependent reductase enzymes will be systematically varied at amino acid positions rated most likely to yield improved mutants. X-ray crystallography will be used to understand how substrates bind in the active sites of improved enzymes.The broader impacts involve mentoring high school students (U.F. Precollegiate Training Program), undergraduates (NSF REU), graduate students and postdoctoral researchers, including members of historically underrepresented groups in chemistry. Moreover, the work will provide diverse training in chemistry, x-ray crystallography, molecular biology and biochemistry to these coworkers. In terms of scientific broader impacts, this work is directed at efficient, environmentally benign methods for producing chiral building blocks for medicinal, agrochemical and industrial chemical applications. In addition, the insights gleaned from combining protein engineering, structural studies and catalytic assays will provide new information on flavoproteins, an important class of biocatalysts involved in a diverse set of reactions at the chemistry/biology interface.