As a natural sesquiterpene compound with numerous biological activities, α-santalene has extensive applications in the cosmetic and pharmaceutical industries. Although several α-santalene-producing microbial strains have been constructed, low productivity still hampers large-scale fermentation. Herein, we present a case of engineered sesquiterpene biosynthesis where the insufficient downstream pathway capacity limited high-level α-santalene production in Escherichia coli. The initial strain was constructed, and it produced 6.4 mg/L α-santalene. To increase α-santalene biosynthesis, we amplified the flux toward farnesyl diphosphate (FPP) precursor by screening and choosing the right FPP synthase and reprogrammed the rate-limiting downstream pathway by generating mutations in santalene synthase (Clausena lansium; ClSS). Santalene synthase was engineered by site-directed mutagenesis, resulting in the improved soluble expression of ClSS and an α-santalene titer of 887.5 mg/L; the α-santalene titer reached 1078.8 mg/L after adding a fusion tag to ClSS. The most productive pathway, which included combining precursor flux amplification and mutant synthases, conferred an approximate 169-fold increase in α-santalene levels. Maximum titers of 1272 and 2916 mg/L were achieved under shake flask and fed-batch fermentation, respectively, and were among the highest levels reported using E. coli as the host.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.