SusChEM: Small-Molecule Enantiopurity Measurement in Living Cells as a Method to Accelerate Biocatalyst Discovery

国家科学基金会化学部化学测量和成像计划为该项目提供资金。犹他大学的詹妮弗·希姆斯特拉教授正在开发快速分析反应条件的方法，以选择性地生产小分子。这项研究有助于使用酶来催化治疗药物和其他高价值化学品的合成中的关键步骤。目前用于生成这些产品的反应通常需要含金属的催化剂和有机溶剂。催化剂和溶剂的处置都会对环境造成危害。相比之下，酶催化剂是环保的，因为酶本身是无毒的，而且它们使用在水中，而不是有机溶剂。这项研究的更广泛的影响通过多种方式得到证明。这项研究为高中、本科生和研究生提供培训。还包括一项旨在改善大学女生在STEM学科中留住的外联活动。这项研究通过促进发现新的、对环境无害的方法来合成高价值化学品(如小分子疗法)，从而有可能造福社会。这项提议的首要目标是探索快速产生用于感兴趣的小分子靶标的DNA生物传感器的方法，并将这些传感器部署在活的细菌细胞中。开发了能够选择用于酶产生的小分子的DNA生物传感器的方法，所述小分子是药物中间体。这项拟议的研究探索了将荧光DNA生物传感器导入细菌细胞并用于量化目标分析物的细胞内浓度的能力。

The Chemical Measurement and Imaging program of the Chemistry Division of the National Science Foundation funds this project. Professor Jennifer Heemstra of the University of Utah is developing methods to rapidly assay reaction conditions for the selective production of small molecules. This research aids in the use of enzymes to catalyze key steps in the synthesis of therapeutics and other high value chemicals. The reactions currently used to generate these products often require metal-containing catalysts and organic solvents. The disposal of both catalyst and solvent pose an environmental hazard. In contrast, enzyme catalysts are environmentally benign, as the enzymes themselves are non-toxic, and they are used in water rather than organic solvents. The broader impacts of this research are demonstrated in multiple ways. This research provides training to high school, undergraduate, and graduate students. Also included is an outreach activity aimed at improving the retention of college women in STEM disciplines. The research has potential to benefit society by facilitating the discovery of new, environmentally benign methods for the synthesis of high-value chemicals such as small-molecule therapeutics.The overarching objectives of this proposal are to explore methods to rapidly generate DNA biosensors for small-molecule targets of interest and to deploy these sensors in living bacterial cells. Methods are developed to enable the selection of DNA biosensors for an enzymatically-produced small molecule that is a pharmaceutical intermediate. The proposed research explores the ability of fluorescent DNA biosensors to be transfected into bacterial cells and used to quantify the intracellular concentration of a target analyte.