Two-step pathway for isoprenoid synthesis

Significance Isoprenoids are a class of natural products with applications in many fields, including medicine and agriculture. Biosynthesis of isoprenoids has emerged as the most commercially viable option for their mass production. We report the creation of an enzymatic pathway for the biological production of isoprenoids, which uses externally provided isoprenol as its substrate instead of a glucose-derived catabolite, making it radically different from naturally occurring pathways or their engineered variants. The pathway is only two steps long and uses a single cofactor, ATP. Our results show that it is decoupled from central carbon metabolism and can sustain a very high flux. These advantages make it a noteworthy alternative to known isoprenoid pathways.