title: Engineering a functional 1-deoxy-D-xylulose 5-phosphate (DXP) pathway in Saccharomyces cerevisiae.
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.
authors.affiliation: California Institute of Quantitative Biosciences (QB3), University of California, Berkeley, CA 94702, USA; Joint BioEnergy Institute, Emeryville, CA 94608, USA.
authors.forename: James
authors.initials: J
authors.lastname: Kirby
authors.affiliation: Amyris, Inc., 5885 Hollis Street, Suite 100, Emeryville, CA 94608, USA.
authors.forename: Kevin L
authors.initials: KL
authors.lastname: Dietzel
authors.affiliation: Amyris, Inc., 5885 Hollis Street, Suite 100, Emeryville, CA 94608, USA.
authors.forename: Gale
authors.initials: G
authors.lastname: Wichmann
authors.affiliation: California Institute of Quantitative Biosciences (QB3), University of California, Berkeley, CA 94702, USA; Joint BioEnergy Institute, Emeryville, CA 94608, USA.
authors.forename: Rossana
authors.initials: R
authors.lastname: Chan
authors.affiliation: Amyris, Inc., 5885 Hollis Street, Suite 100, Emeryville, CA 94608, USA.
authors.forename: Eugene
authors.initials: E
authors.lastname: Antipov
authors.affiliation: Amyris, Inc., 5885 Hollis Street, Suite 100, Emeryville, CA 94608, USA.
authors.forename: Nathan
authors.initials: N
authors.lastname: Moss
authors.affiliation: Joint BioEnergy Institute, Emeryville, CA 94608, USA.
authors.forename: Edward E K
authors.initials: EEK
authors.lastname: Baidoo
authors.affiliation: Amyris, Inc., 5885 Hollis Street, Suite 100, Emeryville, CA 94608, USA.
authors.forename: Peter
authors.initials: P
authors.lastname: Jackson
authors.affiliation: Amyris, Inc., 5885 Hollis Street, Suite 100, Emeryville, CA 94608, USA.
authors.forename: Sara P
authors.initials: SP
authors.lastname: Gaucher
authors.affiliation: Amyris, Inc., 5885 Hollis Street, Suite 100, Emeryville, CA 94608, USA.
authors.forename: Shayin
authors.initials: S
authors.lastname: Gottlieb
authors.affiliation: Amyris, Inc., 5885 Hollis Street, Suite 100, Emeryville, CA 94608, USA.
authors.forename: Jeremy
authors.initials: J
authors.lastname: LaBarge
authors.affiliation: Amyris, Inc., 5885 Hollis Street, Suite 100, Emeryville, CA 94608, USA.
authors.forename: Tina
authors.initials: T
authors.lastname: Mahatdejkul
authors.affiliation: Amyris, Inc., 5885 Hollis Street, Suite 100, Emeryville, CA 94608, USA.
authors.forename: Kristy M
authors.initials: KM
authors.lastname: Hawkins
authors.affiliation: Amyris, Inc., 5885 Hollis Street, Suite 100, Emeryville, CA 94608, USA.
authors.forename: Sheela
authors.initials: S
authors.lastname: Muley
authors.affiliation: Amyris, Inc., 5885 Hollis Street, Suite 100, Emeryville, CA 94608, USA.
authors.forename: Jack D
authors.initials: JD
authors.lastname: Newman
authors.affiliation: Department of Chemistry, Boston University, Boston, MA 02215, USA.
authors.forename: Pinghua
authors.initials: P
authors.lastname: Liu
authors.affiliation: California Institute of Quantitative Biosciences (QB3), University of California, Berkeley, CA 94702, USA; Joint BioEnergy Institute, Emeryville, CA 94608, USA; Departments of Chemical & Biomolecular Engineering and of Bioengineering, University of California, Berkeley, CA 94702, USA; Biological Systems & Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94702, USA; Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kogle Allé, DK2970 Hørsholm, Denmark.
authors.forename: Jay D
authors.initials: JD
authors.lastname: Keasling
authors.affiliation: Amyris, Inc., 5885 Hollis Street, Suite 100, Emeryville, CA 94608, USA. Electronic address: zhao@amyris.com.
authors.forename: Lishan
authors.initials: L
authors.lastname: Zhao
mesh_terms: D060847:Metabolic Engineering; D010428:Pentosephosphates / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D013729:Terpenes / Q000378:metabolism*
chemical_list: C109460:1-deoxylulose 5-phosphate; D010428:Pentosephosphates; D013729:Terpenes
keywords: MEP pathway; metabolic engineering; terpene; yeast