Engineering Streptomyces for Efficient Terpenoid Production

Abstract

Terpenoids are structurally complex natural products with broad agricultural and therapeutic value, yet their low natural abundance and oxidative tailoring requirements complicate scalable production. Conventional microbial hosts often fail to support cytochrome P450-mediated oxidative tailoring, limiting access to complex derivatives. Here, we developed two Streptomyces hosts as alternative chassis, leveraging their extensive biosynthetic capabilities and native oxidative enzyme repertoires. Through the EcoMine strategy, we identified and assembled a translationally coupled TriMEP cassette containing key rate-limiting MEP pathway genes from a high-producing strain and expressed it under strong constitutive promoters. Medium optimization enabled high-level production of four terpenoids, including three oxidized products, in shake-flask cultures: (−)-epi-α-bisabolol (1, 237 ± 14 mg/L), ent-(13Z)-isocupressic acid (2, 173 ± 14 mg/L), ent-(13E)-isocupressic acid (3, 209 ± 20 mg/L), and ent-atiserenoic acid (4, 325 ± 17 mg/L). These results highlight Streptomyces as a robust and underutilized chassis for sustainable production of complex terpenoids.