Improving Alkene Reductases for Applications in Asymmetric Synthesis

有了这个奖项，化学合成计划是支持教授乔恩D。斯图尔特的化学系在佛罗里达大学开发新的酶催化剂的不对称共轭烯烃还原。 迄今为止，这些酶对合成化学的总体影响是适度的，因为野生型酶仅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，现有的黄素依赖性还原酶将在最有可能产生改进的突变体的氨基酸位置上系统地改变。 X射线晶体学将用于了解底物如何结合在改进的酶的活性位点。大学前培训计划），本科生（NSF REU），研究生和博士后研究人员，包括化学历史上代表性不足的群体的成员。 此外，这项工作将为这些同事提供化学，X射线晶体学，分子生物学和生物化学方面的各种培训。 在更广泛的科学影响方面，这项工作是针对高效，环境友好的方法，用于生产医药，农业化学品和工业化学品应用的手性构件。 此外，从结合蛋白质工程，结构研究和催化分析中收集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，参与化学/生物学界面的各种反应。

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.