Producing Gram-Scale Unnatural Rosavin Analogues from Glucose by Engineered Escherichia Coli

Cinnamyl alcohol glycosides (CAGs) are key active ingredients of the precious medicinal plant Rhodiola rosea L, which has diverse pharmacological activities. The quality of R. rosea extracts is standardized to the contents of rosavin, a cinnamyl alcohol disaccharide, along with salidroside. The supply of rosavin and analogues is limited by both the inefficiency of chemical synthesis methods and the shortage of natural resources. Herein, we achieved de novo synthesis of a series of rosavin analogues by engineered Escherichia coli strains. First, cinnamyl alcohol was synthesized by expression of phenylalanine ammonia-lyase (PAL), hydroxycinnamate:CoA ligase, and cinnamyl-CoA reductase in a phenylalanine high-producing strain. UGT73C5 from Arabidopsis thaliana and a sugar chain elongating glycosyltransferase from Catharanthus roseus, CaUGT3 sequentially catalyzed the formation of an unnatural cinnamyl alcohol diglucoside, named rosavin B. Then, these biosynthetic enzymes were transformed into a tyrosine high...