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- W2076870358 abstract "Forskolin, a labdane diterpenoid, possesses a wide range of pharmacological activities. (13R)-manoyl oxide (13R-MO) is the precursor of forskolin. As forskolin is a structurally complex, highly oxidized compound, chemical synthesis is tedious and difficult. Herein, we present a biosynthesis method involving metabolic engineering of Saccharomyces cerevisiae to produce 13R-MO at an initial titer of 2.3 mg/l. We further optimized the entire MVA pathway, which increased the FPP supply pool, but resulted in a sharp decrease in the 13R-MO production due to low metabolic flux toward geran ylgeranyl pyrophosphate (GGPP). To mitigate this, we down-regulated the competing pathway by replacing the original promoter of the squalene synthase gene ERG9 with the MET3 promoter, significantly improving the 13R-MO production to 45.2 mg/l. Finally, three feed strategies were investigated in fed-batch fermentation; the glucose feeding strategy enabled the engineered yeast to produce 167.1 ± 5.2 mg/l 13R-MO, equivalent to 2.6 ± 0.08 mg/l/OD600. Here, we describe a systematic synthetic biology method to produce 13R-MO from a simple carbon source using Saccharomyces cerevisiae. This not only enabled a relatively high level of 13R-MO production but also provided an efficient platform for the production of other diterpenoids." @default.
- W2076870358 created "2016-06-24" @default.
- W2076870358 creator A5013176506 @default.
- W2076870358 creator A5019059825 @default.
- W2076870358 creator A5048681901 @default.
- W2076870358 date "1992-01-01" @default.
- W2076870358 modified "2023-10-14" @default.
- W2076870358 title "Synthetic routes to forskolin" @default.
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- W2076870358 doi "https://doi.org/10.1016/s0040-4020(01)88199-0" @default.
- W2076870358 hasPublicationYear "1992" @default.
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