Multi-substrate conversion synchronous cofactor-regeneration for efficient natural product biosynthesis

Abstract

Redox cofactors are pivotal electron mediators in natural product biosynthesis, yet the sacrificial agents required for their regeneration compromise atomic economy and generate unwanted by-products. Herein, we propose a multi-substrate conversion synchronous cofactor-regeneration strategy (MSCR) to eliminate the need for sacrificial agents, which establishes a bidirectional redox balance between oxidizing and reducing substrates. Based on MSCR, we designed a three-enzyme cascade to convert inexpensive substrates rhododendrol and 4-hydroxybenzylideneacetone into raspberry ketone. Through enzyme engineering and activity balancing, raspberry ketone was obtained without sacrificial agents, achieving a carbon atom conversion rate of 94.68%. Given the widespread natural abundance of reducing and oxidizing precursors, MSCR strategy holds significant potential for extension to access a vast array of valuable natural products.