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

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.