Metabolic engineering of Escherichia coli for the biosynthesis of α-copaene from glucose

α-Copaene, a tricyclic sesquiterpene, has broad application prospects in many fields, including food, medicine, and agriculture. The conventional plant extraction method of α-copaene is unsustainable. In this study, for the first time, the de novo biosynthesis of α-copaene from glucose was achieved using a metabolically engineered Escherichia coli strain. α-Copaene was successfully produced in the engineered strain that heterologously expressed Piper nigrum α-copaene synthase. A series of strategies were employed to enhance α-copaene biosynthesis by E. coli , including heterologous overexpression of Saccharomyces cerevisiae farnesyl diphosphate synthase, MVA pathway, and truncated 3-hydroxy-3-methylglutaryl coenzyme A reductase 1. Strain optimization led to a 78.1-fold increase in α-copaene level in E. coli . This study reports a sustainable method for α-copaene production and provides a metabolic engineering strategy for the biosynthesis of other sesquiterpenes in E. coli . • α-Copaene was de novo biosynthesized via metabolic engineered E. coli. • PnTPS3, ERG20, and mevalonate pathway genes were expressed in E. coli. • Strain optimization led to a 78.1-fold increase in α-copaene level. • Establishment of a sustainable method for α-copaene production.