Highly Efficient Biosynthesis of γ-Bisabolene with a New Sesquiterpene Synthase AcTPS5 by Dual Cytoplasmic-Peroxisomal Engineering in Saccharomyces cerevisiae
2023
Jiajia Liu | Ge Yao | Xiukun Wan | Fuli Wang | Penggang Han | Shaoheng Bao | Kang Wang | Tianyu Song | Hui Jiang
&gamma:-bisabolene is a monocyclic sesquiterpene with various biological activities: it has also been approved as a food additive. Additionally, the hydrogenated form of bisabolene is considered as a potential alternative to D2 diesel. Saccharomyces cerevisiae has the ability to produce a large amount of acetyl-CoA in both cytosol and peroxisomes, which serves as a precursor in terpene biosynthesis. In this study, AcTPS5 was identified as a new &gamma:-bisabolene synthase. By expressing AcTPS5 and the mevalonate pathway in peroxisomes, &gamma:-bisabolene titer was achieved at 125.0 mg/L. Deleting the peroxisome autophagy gene atg36 further improved &gamma:-bisabolene production to 216.9 mg/L. The implementation of dual cytoplasmic&ndash:peroxisomal engineering further boosted &gamma:-bisabolene production to 296.4 mg/L. Finally, through increasing the acetyl-CoA supply and down-regulating the expression of ERG9, &gamma:-bisabolene production was achieved at 584.14 mg/L in shake-flask fermentation and 2.69 g/L in fed-batch fermentation, which is the highest reported production of &gamma:-bisabolene to date. The strategy presented in this study provides an efficient approach for terpene production in S. cerevisiae.
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