Elevating Limonene Production in Oleaginous Yeast Yarrowia lipolytica via Genetic Engineering of Limonene Biosynthesis Pathway and Optimization of Medium Composition
- 29 Downloads
As an alternative terpenoid producer, non-conventional oleaginous yeast Yarrowia lipolytica was chosen for limonene production. Y. lipolytica can give high biomass yield and metabolize a broad range of substrates such as glycerol, alkanes, fatty acid, fats, and oils. As previously reported, optimization of limonene synthesis pathway and mevalonate (MVA) pathway leads to the accumulation of 112-fold higher limonene as compared to an initial strain. In this study, we introduced an additional copy of limonene synthesis gene (LS), which resulted in an increase of limonene production. This engineered strain was used to carry out further optimization study. Amongst all the carbon sources tested, the highest level of limonene production was obtained from glycerol, and citrate was selected as an auxiliary carbon source. In fed-batch fermentation with an optimized medium, the engineered strain was found to produce 165.3 mg/L limonene, which corresponds to the highest yield till date for the production of limonene in Y. lipolytica.
Keywordslimonene Yarrowia lipolytica carbon source fed-batch fermentation
Unable to display preview. Download preview PDF.
This study was financially supported by National Natural Science Foundation of China (21776081, 21576089) and Natural Science Foundation of Shanghai (18ZR1410000).
- 1.Sun, J. (2007) D-limonene: Safety and clinical applications. Altern. Med. Rev. 12: 259–264.Google Scholar
- 7.Jongedijk, E., K. Cankar, J. Ranzijn, S. van der Krol, H. Bouwmeester, and J. Beekwilder (2015) Capturing of themonoterpene olefin limonene produced in Saccharomyces cerevisiae. Yeast 32: 159–171.Google Scholar
- 8.Alonso-Gutierrez, J., E. M. Kim, T. S. Batth, N. Cho, Q. J. Hu, L. J. G. Chan, C. J. Petzold, N. J. Hinson, P. D. Adams, J. D. Keasling, H. G. Martin, and T. S. Lee (2015) Principal component analysis of proteomics (PCAP) as a tool to direct metabolic engineering. Metab. Eng. 28: 123–133.CrossRefGoogle Scholar
- 14.Michely, S., C. Gaillardin, J. M. Nicaud, and C. Neuveglise (2013) Comparative physiology of oleaginous species from the Yarrowia clade. PLoS One 8.Google Scholar
- 17.Kim, J. H., S. W. Kim, D. Q. A. Nguyen, H. Li, S. B. Kim, Y. G. Seo, J. K. Yang, I. Y. Chung, D. H. Kim, and C. J. Kim (2009) Production of beta-carotene by recombinant Escherichia coli with engineered whole mevalonate pathway in batch and fed-batch cultures. Biotechnol. Bioproc. E. 14: 559–564.CrossRefGoogle Scholar