Abstract
Yarrowia lipolytica is a well-known oleaginous yeast that naturally accumulates lipids to more than 20% of their dry cell weight. Due to its brief doubling time and Generally Recognized as Safe (GRAS) properties, Y. lipolytica has been exploited for the production of commercially valuable lipids. Among the genes related to the lipid synthesis, the gene YALI0E16797g (LRO1) encoding a major triacylglycerol synthase of Y. lipolytica shows a significant impact during the acylation process. Thus, in the present work, we explore the contributions of hp4d or TEFintron promoters to the response of LRO1 expression on lipid accumulation by molecular cloning technology. Results showed that over-expression of LRO1 led to higher lipid content as well as lipid yield. The one with the hp4d promoter showed the highest lipid content of 12% wt. However, such an enhancement also caused a growth defect of cells. On the other hand, the lipid content of the cells over-expressing LRO1 with TEFintron promoter revealed only a modest increase in lipid content, but it promoted cell growth. Therefore, all things considered the one with the TEFintron promoter showed the highest lipid yield.
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References
Alper, H., & Stephanopoulos, G. (2009). Nature Reviews. Microbiology, 7, 715–723.
Beopoulos, A., Nicaud, J. M., & Gaillardin, C. (2011). An overview of lipid metabolism in yeasts and its impact on biotechnological processes. Applied Microbiology and Biotechnology, 90(4), 1193–1206.
Karmakar, A., Karmakar, S., & Mukherjee, S. (2010). Properties of various plants and animals feedstocks for biodiesel production. Bioresource Technology, 101(19), 7201–7210.
Windahl, K., Faxen Irving, G., Almquist, T., Liden, M. K., van de Luijtgaarden, M., Chesnaye, N. C., Voskamp, P., Stenvinkel, P., Klinger, M., Szymczak, M., Torino, C., Postorini, M., Drechsler, C., Caskey, F. J., Wanner, C., Dekker, F. W., Jager, K. J., & Evans, M. (2018). Journal of Renal Nutrition, 28, 165–174.
Zhu, Q., & Jackson, E. N. (2015). Metabolic engineering of Yarrowia lipolytica for industrial applications. Current Opinion in Biotechnology, 36, 65–72.
Gibellini, F., & Smith, T. K. (2010). IUBMB Life, 62, 414–428.
Beopoulos, A., Chardot, T., & Nicaud, J. M. (2009). Yarrowia lipolytica: a model and a tool to understand the mechanisms implicated in lipid accumulation. Biochimie, 91(6), 692–696.
Oelkers, P., Tinkelenberg, A., Erdeniz, N., Cromley, D., Billheimer, J. T., & Sturley, S. L. (2000). A lecithin cholesterol acyltransferase-like gene mediates diacylglycerol esterification in yeast. The Journal of Biological Chemistry, 275(21), 15609–15612.
Sorger, D., Athenstaedt, K., Hrastnik, C., & Daum, G. (2004). A yeast strain lacking lipid particles bears a defect in ergosterol formation. The Journal of Biological Chemistry, 279(30), 31190–31196.
Dahlqvist, A., Stahl, U., Lenman, M., Banas, A., Lee, M., Sandager, L., Ronne, H., & Stymne, S. (2000). Phospholipid:diacylglycerol acyltransferase: an enzyme that catalyzes the acyl-CoA-independent formation of triacylglycerol in yeast and plants. Proceedings of the National Academy of Sciences of the United States of America, 97(12), 6487–6492.
Banas, A., Dahlqvist, A., Stahl, U., Lenman, M., & Stymne, S. (2000). The involvement of phospholipid:diacylglycerol acyltransferases in triacylglycerol production. Biochemical Society Transactions, 28(6), 703–705.
Tai, M., & Stephanopoulos, G. (2013). Metabolic Engineering, 15, 1–9.
Courchesne, N. M., Parisien, A., Wang, B., & Lan, C. Q. (2009). Enhancement of lipid production using biochemical, genetic and transcription factor engineering approaches. Journal of Biotechnology, 141(1-2), 31–41.
Athenstaedt, K. (2011). YALI0E32769g (DGA1) and YALI0E16797g (LRO1) encode major triacylglycerol synthases of the oleaginous yeast Yarrowia lipolytica. Biochimica et Biophysica Acta, 1811(10), 587–596.
Zhang, H., Damude, H. G., & Yadav, N. S. (2012). Three diacylglycerol acyltransferases contribute to oil biosynthesis and normal growth in Yarrowia lipolytica. Yeast, 29(1), 25–38.
Madzak, C., Treton, B., & Blanchin-Roland, S. (2000). Journal of Molecular Microbiology and Biotechnology, 2, 207–216.
Le Hir, H., Nott, A., & Moore, M. J. (2003). Trends in Biochemical Sciences, 28, 215–220.
Nevoigt, E., Kohnke, J., Fischer, C. R., Alper, H., Stahl, U., & Stephanopoulos, G. (2006). Applied and Environmental Microbiology, 72, 5266–5273.
Li, Y. Q., Horsman, M., Wang, B., Wu, N., & Lan, C. Q. (2008). Applied Microbiology and Biotechnology, 81, 629–636.
Wahidin, S., Idris, A., & Shaleh, S. R. M. (2013). Bioresource Technology, 129, 7–11.
Papanikolaou, S., & Aggelis, G. (2002). Lipid production by Yarrowia lipolytica growing on industrial glycerol in a single-stage continuous culture. Bioresource Technology, 82(1), 43–49.
Acknowledgements
This work was supported by the Ministry of Science and Technology, Taiwan (grant numbers MOST 107-2221-E-011-002-MY3 and MOST 105-2221-E-011 -002 -MY3).
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Amalia, L., Zhang, YH., Ju, YH. et al. Enhanced Lipid Production in Yarrowia lipolytica Po1g by Over-expressing lro1 Gene under Two Different Promoters. Appl Biochem Biotechnol 191, 104–111 (2020). https://doi.org/10.1007/s12010-020-03226-9
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DOI: https://doi.org/10.1007/s12010-020-03226-9