Rational Reprogramming of <i>O</i>‑Methylation Regioselectivity for Combinatorial Biosynthetic Tailoring of Benzenediol Lactone Scaffolds Xiaojing Wang Chen Wang Lixin Duan Liwen Zhang Hang Liu Ya-ming Xu Qingpei Liu Tonglin Mao Wei Zhang Ming Chen Min Lin A. A. Leslie Gunatilaka Yuquan Xu István Molnár 10.1021/jacs.8b12967.s001 https://acs.figshare.com/articles/journal_contribution/Rational_Reprogramming_of_i_O_i_Methylation_Regioselectivity_for_Combinatorial_Biosynthetic_Tailoring_of_Benzenediol_Lactone_Scaffolds/7776380 <i>O-</i>Methylation modulates the pharmaco­kinetic and pharmaco­dynamic (PK/PD) properties of small-molecule natural products, affecting their bioavailability, stability, and binding to targets. Diversity-oriented combinatorial biosynthesis of new chemical entities for drug discovery and optimization of known bioactive scaffolds during drug development both demand efficient <i>O-</i>methyl­transferase (OMT) biocatalysts with considerable substrate promiscuity and tunable regio­selectivity that can be deployed in a scalable and sustainable manner. Here we demonstrate efficient total biosynthetic and biocatalytic platforms that use a pair of fungal OMTs with orthogonal regio­specificity to produce unnatural <i>O-</i>methylated benzenediol lactone polyketides. We show that rational, structure-guided active-site cavity engineering can reprogram the regio­selectivity of these enzymes. We also characterize the interplay of engineered regio­selectivity with substrate plasticity. These findings will guide combinatorial biosynthetic tailoring of unnatural products toward the generation of diverse chemical matter for drug discovery and the PK/PD optimization of bioactive scaffolds for drug development. 2019-02-15 00:00:00 pharmaco drug discovery biosynthetic regio methylated benzenediol lactone polyketides substrate drug development PK Benzenediol Lactone Scaffolds O bioactive scaffolds Diversity-oriented combinatorial biosynthesis Combinatorial Biosynthetic Tailoring optimization OMT chemical Methylation structure-guided active-site cavity engineering