Molecular basis for enzymatic control of enantiodivergent Diels-Alder reactions in spirotetronate biosynthesis

在医药、动物保健和农业中应用的复杂分子的化学合成往往效率低下和繁琐，存在的问题包括总体产品产量低、扩大规模困难以及严重依赖不可再生资源。利用天然产物生物合成酶是一种更可持续、可扩展和高效生产复杂分子的有前途的策略。然而，要实现这一目标，需要对这些酶的催化机制进行详细的分子理解。该项目旨在更好地了解在螺戊酸天然产物生物合成中催化关键[4+2]环加成反应的diols - alderase酶如何控制其产物的立体化学。将采用一种高度跨学科的方法，将底物和底物类似物的合成与酶学、分析化学、结构生物学、定点诱变和对接模拟相结合。这项工作的结果将为未来开发催化复杂性生成转化的酶奠定基础，例如Diels-Alder[4+2]环加成，以更可持续地生产结构复杂的分子。

Chemical synthesis of complex molecules with applications in medicine, animal health and agriculture is often inefficient and cumbersome, with problems including poor overall product yields, difficulties with scale-up and heavy reliance on non-renewable resources. Harnessing natural product biosynthetic enzymes is a promising strategy for more sustainable, scalable and efficient production of complex molecules. However, a detailed molecular understanding of the catalytic mechanisms of such enzymes is required to achieve this. This project aims to develop a better understanding of how Diels-Alderase enzymes that catalyse key [4+2] cycloaddition reactions in the biosynthesis of spirotetronate natural products control the stereochemistry of their products. A highly interdisciplinary approach combining synthesis of substrates and substrate analogues with enzymology, analytical chemistry, structural biology, site-directed mutagenesis and docking simulations will be employed . The outcomes of this work will underpin future efforts to exploit enzymes that catalyse complexity-generating transformations, such as Diels-Alder [4+2] cycloadditions, for more sustainable production of structurally complex molecules.