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- W4247020854 abstract "Abstract Background The sesquiterpene germacrene A is a direct precursor of ß-elemene that is a major component of the Chinese medicinal herb Curcuma wenyujin with prominent antitumor activity. The microbial platform for germacrene A production was previously established in Saccharomyces cerevisiae using the germacrene A synthase (LTC2) of Lactuca sativa . Results We evaluated the performance of LTC2 as well as nine other identified or putative germacrene A synthases from different sources for the production of germacrene A. Av GAS, a synthase of Anabaena variabilis , was found to be the most efficient in germacrene A production in yeast. Av GAS expression alone in S. cerevisiae CEN.PK2-1D already resulted in a substantial production of germacrene A while LTC2 expression did not. Further metabolic engineering the yeast using known strategies including overexpression of tHMGR1, repression of squalene synthesis pathway and decreasing farnesol synthesis, led to an 11-fold increase in germacrene A production. Site-directed mutagenesis of Av GAS revealed that while changes of several residues located within the active site cavity severely compromised germacrene A production , substitution of Phe23 located on the lateral surface with tryptophan or valine led to a 35.2% and 21.8% increase in germacrene A production, respectively. Finally, the highest production titer of germacrene A reached 309.8 mg/L in shake-flask batch culture. Conclusions Our study highlights the potential of applying bacterial sesquiterpene synthases with improved performance by mutagenesis engineering in producing germacrene A." @default.
- W4247020854 created "2022-05-12" @default.
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- W4247020854 date "2020-10-01" @default.
- W4247020854 modified "2023-10-16" @default.
- W4247020854 title "Improved Production of Germacrene A, a Direct Precursor of ß-elemene, in Engineered Saccharomyces Cerevisiae by Expressing a Cyanobacterial Germacrene A Synthase " @default.
- W4247020854 doi "https://doi.org/10.21203/rs.3.rs-80968/v1" @default.
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