Engineering a functional 1-deoxy-D-xylulose 5-phosphate (DXP) pathway in Saccharomyces cerevisiae

James Kirby, Kevin L. Dietzel, Gale Wichmann, Rossana Chan, Eugene Antipov, Nathan Moss, Edward E.K. Baidoo, Peter Jackson, Sara P. Gaucher, Shayin Gottlieb, Jeremy LaBarge, Tina Mahatdejkul, Kristy M. Hawkins, Sheela Muley, Jack D. Newman, Pinghua Liu, Jay Keasling, Lishan Zhao

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

Isoprenoids are used in many commercial applications and much work has gone into engineering microbial hosts for their production. Isoprenoids are produced either from acetyl-CoA via the mevalonate pathway or from pyruvate and glyceraldehyde 3-phosphate via the 1-deoxy-D-xylulose 5-phosphate (DXP) pathway. Saccharomyces cerevisiae exclusively utilizes the mevalonate pathway to synthesize native isoprenoids and in fact the alternative DXP pathway has never been found or successfully reconstructed in the eukaryotic cytosol. There are, however, several advantages to isoprenoid synthesis via the DXP pathway, such as a higher theoretical yield, and it has long been a goal to transplant the pathway into yeast. In this work, we investigate and address barriers to DXP pathway functionality in S. cerevisiae using a combination of synthetic biology, biochemistry and metabolomics. We report, for the first time, functional expression of the DXP pathway in S. cerevisiae. Under low aeration conditions, an engineered strain relying solely on the DXP pathway for isoprenoid biosynthesis achieved an endpoint biomass 80% of that of the same strain using the mevalonate pathway.
Original languageEnglish
JournalMetabolic Engineering
Volume38
Pages (from-to)494-503
Number of pages10
ISSN1096-7176
DOIs
Publication statusPublished - 2016

Keywords

  • MEP pathway
  • Yeast
  • Metabolic engineering
  • Terpene

Cite this

Kirby, J., Dietzel, K. L., Wichmann, G., Chan, R., Antipov, E., Moss, N., Baidoo, E. E. K., Jackson, P., Gaucher, S. P., Gottlieb, S., LaBarge, J., Mahatdejkul, T., Hawkins, K. M., Muley, S., Newman, J. D., Liu, P., Keasling, J., & Zhao, L. (2016). Engineering a functional 1-deoxy-D-xylulose 5-phosphate (DXP) pathway in Saccharomyces cerevisiae. Metabolic Engineering, 38, 494-503. https://doi.org/10.1016/j.ymben.2016.10.017