Work

Cell-Free Total Biosynthesis of Plant Terpene Natural Products Using an Orthogonal Cofactor Regeneration System

Year: 2021

Type: article

Abstract: Here we report the one-pot, cell-free enzymatic synthesis of the plant monoterpene nepetalactol starting from the readily available geraniol. A pair of orthogonal cofactor regeneration systems permitted NAD + -dependent geraniol oxidation followed by NADPH-dependent reductive cyclization without isolation of intermediates. The orthogonal cofactor regeneration system maintained a high ratio of NAD + to NADH and a low ratio of NADP + to NADPH. The overall reaction contains four biosynthetic enzymes, including a soluble P450, and five accessory and cofactor regeneration enzymes. Furthermore, addition of a NAD + -dependent dehydrogenase to the one-pot mixture led to ∼1 g/L of nepetalactone, the active cat attractant in catnip.

Source: ACS Catalysis

DOI: 10.1021/acscatal.1c02267