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

Abstract

Isoprenoids are made by all free-living organisms and range from essential metabolites like sterols and quinones to more complex compounds like pinene and rubber. They 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.

Authors:
 [1];  [2];  [2];  [1];  [2];  [2];  [3];  [2];  [2];  [2];  [2];  [2];  [2];  [2];  [2];  [4];  [5];  [2]
+ Show Author Affiliations
  1. Univ. of California, Berkeley, CA (United States). California Institute of Quantitative Biosciences (QB3); Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States)
  2. Amyris, inc., Emeryville, CA (United States)
  3. Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States)
  4. Boston Univ., MA (United States). Dept. of Chemistry
  5. Univ. of California, Berkeley, CA (United States). California Institute of Quantitative Biosciences (QB3); Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Univ. of California, Berkeley, CA (United States). Depts. of Chemical & Biomolecular Engineering and Bioengineering; Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Biological Systems & Engineering Div.; Technical Univ. of Denmark, Hoesholm (Denmark). Novo Nodisk Foundation Center for Biosustainability
Publication Date:
Research Org.:
Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER)
OSTI Identifier:
1378361
Alternate Identifier(s):
OSTI ID: 1397055
Grant/Contract Number:  
AC02-05CH11231; 180561; GM093903
Resource Type:
Accepted Manuscript
Journal Name:
Metabolic Engineering
Additional Journal Information:
Journal Volume: 38; Journal Issue: C; Journal ID: ISSN 1096-7176
Publisher:
Elsevier
Country of Publication:
United States
Language:
English
Subject:
36 MATERIALS SCIENCE; 59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Kirby, James, Dietzel, Kevin L., Wichmann, Gale, Chan, Rossana, Antipov, Eugene, Moss, Nathan, Baidoo, Edward E. K., Jackson, Peter, Gaucher, Sara P., Gottlieb, Shayin, LaBarge, Jeremy, Mahatdejkul, Tina, Hawkins, Kristy M., Muley, Sheela, Newman, Jack D., Liu, Pinghua, Keasling, Jay D., and Zhao, Lishan. Engineering a functional 1-deoxy-D-xylulose 5-phosphate (DXP) pathway in Saccharomyces cerevisiae. United States: N. p., 2016. Web. doi:10.1016/j.ymben.2016.10.017.
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Kirby, James, Dietzel, Kevin L., Wichmann, Gale, Chan, Rossana, Antipov, Eugene, Moss, Nathan, Baidoo, Edward E. K., Jackson, Peter, Gaucher, Sara P., Gottlieb, Shayin, LaBarge, Jeremy, Mahatdejkul, Tina, Hawkins, Kristy M., Muley, Sheela, Newman, Jack D., Liu, Pinghua, Keasling, Jay D., & Zhao, Lishan. Engineering a functional 1-deoxy-D-xylulose 5-phosphate (DXP) pathway in Saccharomyces cerevisiae. United States. https://doi.org/10.1016/j.ymben.2016.10.017
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Kirby, James, Dietzel, Kevin L., Wichmann, Gale, Chan, Rossana, Antipov, Eugene, Moss, Nathan, Baidoo, Edward E. K., Jackson, Peter, Gaucher, Sara P., Gottlieb, Shayin, LaBarge, Jeremy, Mahatdejkul, Tina, Hawkins, Kristy M., Muley, Sheela, Newman, Jack D., Liu, Pinghua, Keasling, Jay D., and Zhao, Lishan. Thu . "Engineering a functional 1-deoxy-D-xylulose 5-phosphate (DXP) pathway in Saccharomyces cerevisiae". United States. https://doi.org/10.1016/j.ymben.2016.10.017. https://www.osti.gov/servlets/purl/1378361.
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@article{osti_1378361,
title = {Engineering a functional 1-deoxy-D-xylulose 5-phosphate (DXP) pathway in Saccharomyces cerevisiae},
author = {Kirby, James and Dietzel, Kevin L. and Wichmann, Gale and Chan, Rossana and Antipov, Eugene and Moss, Nathan and Baidoo, Edward E. K. and Jackson, Peter and Gaucher, Sara P. and Gottlieb, Shayin and LaBarge, Jeremy and Mahatdejkul, Tina and Hawkins, Kristy M. and Muley, Sheela and Newman, Jack D. and Liu, Pinghua and Keasling, Jay D. and Zhao, Lishan},
abstractNote = {Isoprenoids are made by all free-living organisms and range from essential metabolites like sterols and quinones to more complex compounds like pinene and rubber. They 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.},
doi = {10.1016/j.ymben.2016.10.017},
journal = {Metabolic Engineering},
number = C,
volume = 38,
place = {United States},
year = {Thu Oct 27 00:00:00 EDT 2016},
month = {Thu Oct 27 00:00:00 EDT 2016}
}
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https://doi.org/10.1016/j.ymben.2016.10.017
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