期刊
JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY
卷 46, 期 2, 页码 147-157出版社
SPRINGER HEIDELBERG
DOI: 10.1007/s10295-018-2116-3
关键词
Triterpenoids; Cucurbitadienol; Metabolic engineering; Saccharomyces cerevisiae
资金
- National Natural Science Foundation of China [81573521]
- Beijing Municipal Natural Science Foundation [5172028]
- CAMS Innovation Fund for Medical Sciences [2017-I2M-1-013]
Cucurbitane-type triterpenoids such as mogrosides and cucurbitacins that are present in the plants of Cucurbitaceae are widely used in Asian traditional medicine. Cucurbitadienol is the skeleton of cucurbitane-type triterpenoids. As an alternative production strategy, we developed baker's yeast Saccharomyces cerevisiae as a microbial host for the eventual transformation of cucurbitadienol. The synthetic pathway of cucurbitadienol was constructed in Saccharomyces cerevisiae by introducing the cucurbitadienol synthase gene from different plants, resulting in 7.80mg cucurbitadienol from 1L of fermentation broth. Improving supplies of isoprenoid precursors was then investigated for increasing cucurbitadienol production. Cucurbitadienol production increased to 21.47mg/L through the overexpression of a global regulatory factor (UPC2) gene of triterpenoid synthase. In addition, knockout of the ERG7 gene increased cucurbitadienol production from 21.47 to 61.80mg/L. Finally, fed-batch fermentation was performed, and 63.00mg/L cucurbitadienol was produced. This work is an important step towards the total biosynthesis of valuable cucurbitane-type triterpenoids and demonstrates the potential for developing a sustainable and secure yeast biomanufacturing platform for triterpenoids.
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