Abstract
BackgroundDiterpenoids are a large class of natural products with complex structures and broad commercial applications as food additives, important medicines, and fragrances. However, their low abundance in plants and high structural complexity limit their applications. Therefore, it is important to create an efficient diterpenoid-producing yeast cell factory of the production of various high-value diterpenoid compounds in a cost-effective mannerResultsIn this study, 13R-manoyl oxide (13R-MO; 2.31 mg/L) was produced by expressing CfTPS2 and CfTPS3 from Coleusforskohlii in Saccharomyces cerevisiae. The 13R-MO titer was increased by 142-fold to 328.15 mg/L via the stepwise metabolic engineering of the original strain, including the overexpression of the rate-limiting genes (tHMG1 and ERG20) of the mevalonate pathway, transcription and protein level regulation of ERG9, Bts1p and Erg20F96Cp fusion, and the overexpression of tCfTPS2 and tCfTPS3 (excision of the N-terminal plastid transit peptide sequences of CfTPS2 and CfTPS3). The final titer of 13R-MO reached up to 3 g/L by fed-batch fermentation in a 5 L bioreactor.ConclusionsIn this study, an efficient 13R-MO yeast cell factory was constructed, which achieved the de novo production of 3 g/L of 13R-MO from glucose. To the best of our knowledge, this is the highest 13R-MO titer reported to date. Furthermore, the metabolic engineering strategies presented here could be used to produce other valuable diterpenoid compounds in yeast.
Highlights
Diterpenoids are a large class of natural products with complex structures and broad commercial applications as food additives, important medicines, and fragrances
The 13R-manoyl oxide (13R-MO) titer was increased by 142-fold to 328.15 mg/L via regulation of the key genes of the MVA pathway and truncation of diterpene synthase (Fig. 1)
Engineering a 13R‐MO synthetic pathway in S. cerevisiae In S. cerevisiae, the diterpenoid precursor geranylgeranyl diphosphate (GGPP) is synthesized from the condensation of farnesyl diphosphate (FPP) and isopentenyl diphosphate (IPP) by Bts1p
Summary
Diterpenoids are a large class of natural products with complex structures and broad commercial applications as food additives, important medicines, and fragrances. Their low abundance in plants and high structural complexity limit their applications. Terpenoids are the main components of plant secondary metabolites, some of which are important to humans as pharmaceuticals and biofuels [2, 3]. Examples of plant-derived terpenoid biofuels include the jet air–fuel additives pinane, Forskolin, a labdane-type diterpenoid, was discovered in the roots of the medicinal shrub Coleus forskohlii and has been used to treat hypertension, heart complications, and asthma [8]. By mining root transcriptome data of C. forskohlii, CfTPS2 which synthesizes
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
