title: Rewriting yeast central carbon metabolism for industrial isoprenoid production.
abstract: A bio-based economy has the potential to provide sustainable substitutes for petroleum-based products and new chemical building blocks for advanced materials. We previously engineered Saccharomyces cerevisiae for industrial production of the isoprenoid artemisinic acid for use in antimalarial treatments. Adapting these strains for biosynthesis of other isoprenoids such as β-farnesene (C15H24), a plant sesquiterpene with versatile industrial applications, is straightforward. However, S. cerevisiae uses a chemically inefficient pathway for isoprenoid biosynthesis, resulting in yield and productivity limitations incompatible with commodity-scale production. Here we use four non-native metabolic reactions to rewire central carbon metabolism in S. cerevisiae, enabling biosynthesis of cytosolic acetyl coenzyme A (acetyl-CoA, the two-carbon isoprenoid precursor) with a reduced ATP requirement, reduced loss of carbon to CO2-emitting reactions, and improved pathway redox balance. We show that strains with rewired central metabolism can devote an identical quantity of sugar to farnesene production as control strains, yet produce 25% more farnesene with that sugar while requiring 75% less oxygen. These changes lower feedstock costs and dramatically increase productivity in industrial fermentations which are by necessity oxygen-constrained. Despite altering key regulatory nodes, engineered strains grow robustly under taxing industrial conditions, maintaining stable yield for two weeks in broth that reaches >15% farnesene by volume. This illustrates that rewiring yeast central metabolism is a viable strategy for cost-effective, large-scale production of acetyl-CoA-derived molecules.
authors.affiliation: Amyris, 5885 Hollis Street, Suite 100, Emeryville, California 94608, USA.
authors.forename: Adam L
authors.initials: AL
authors.lastname: Meadows
authors.affiliation: Amyris, 5885 Hollis Street, Suite 100, Emeryville, California 94608, USA.
authors.forename: Kristy M
authors.initials: KM
authors.lastname: Hawkins
authors.affiliation: Amyris, 5885 Hollis Street, Suite 100, Emeryville, California 94608, USA.
authors.forename: Yoseph
authors.initials: Y
authors.lastname: Tsegaye
authors.affiliation: Amyris, 5885 Hollis Street, Suite 100, Emeryville, California 94608, USA.
authors.forename: Eugene
authors.initials: E
authors.lastname: Antipov
authors.affiliation: Amyris, 5885 Hollis Street, Suite 100, Emeryville, California 94608, USA.
authors.forename: Youngnyun
authors.initials: Y
authors.lastname: Kim
authors.affiliation: Amyris, 5885 Hollis Street, Suite 100, Emeryville, California 94608, USA.
authors.forename: Lauren
authors.initials: L
authors.lastname: Raetz
authors.affiliation: Amyris, 5885 Hollis Street, Suite 100, Emeryville, California 94608, USA.
authors.forename: Robert H
authors.initials: RH
authors.lastname: Dahl
authors.affiliation: Amyris, 5885 Hollis Street, Suite 100, Emeryville, California 94608, USA.
authors.forename: Anna
authors.initials: A
authors.lastname: Tai
authors.affiliation: Amyris, 5885 Hollis Street, Suite 100, Emeryville, California 94608, USA.
authors.forename: Tina
authors.initials: T
authors.lastname: Mahatdejkul-Meadows
authors.affiliation: Amyris, 5885 Hollis Street, Suite 100, Emeryville, California 94608, USA.
authors.forename: Lan
authors.initials: L
authors.lastname: Xu
authors.affiliation: Amyris, 5885 Hollis Street, Suite 100, Emeryville, California 94608, USA.
authors.forename: Lishan
authors.initials: L
authors.lastname: Zhao
authors.affiliation: Amyris, 5885 Hollis Street, Suite 100, Emeryville, California 94608, USA.
authors.forename: Madhukar S
authors.initials: MS
authors.lastname: Dasika
authors.affiliation: Amyris, 5885 Hollis Street, Suite 100, Emeryville, California 94608, USA.
authors.forename: Abhishek
authors.initials: A
authors.lastname: Murarka
authors.affiliation: Amyris, 5885 Hollis Street, Suite 100, Emeryville, California 94608, USA.
authors.forename: Jacob
authors.initials: J
authors.lastname: Lenihan
authors.affiliation: Amyris, 5885 Hollis Street, Suite 100, Emeryville, California 94608, USA.
authors.forename: Diana
authors.initials: D
authors.lastname: Eng
authors.affiliation: Amyris, 5885 Hollis Street, Suite 100, Emeryville, California 94608, USA.
authors.forename: Joshua S
authors.initials: JS
authors.lastname: Leng
authors.affiliation: Amyris, 5885 Hollis Street, Suite 100, Emeryville, California 94608, USA.
authors.forename: Chi-Li
authors.initials: CL
authors.lastname: Liu
authors.affiliation: Amyris, 5885 Hollis Street, Suite 100, Emeryville, California 94608, USA.
authors.forename: Jared W
authors.initials: JW
authors.lastname: Wenger
authors.affiliation: Amyris, 5885 Hollis Street, Suite 100, Emeryville, California 94608, USA.
authors.forename: Hanxiao
authors.initials: H
authors.lastname: Jiang
authors.affiliation: Amyris, 5885 Hollis Street, Suite 100, Emeryville, California 94608, USA.
authors.forename: Lily
authors.initials: L
authors.lastname: Chao
authors.affiliation: Amyris, 5885 Hollis Street, Suite 100, Emeryville, California 94608, USA.
authors.forename: Patrick
authors.initials: P
authors.lastname: Westfall
authors.affiliation: Amyris, 5885 Hollis Street, Suite 100, Emeryville, California 94608, USA.
authors.forename: Jefferson
authors.initials: J
authors.lastname: Lai
authors.affiliation: Amyris, 5885 Hollis Street, Suite 100, Emeryville, California 94608, USA.
authors.forename: Savita
authors.initials: S
authors.lastname: Ganesan
authors.affiliation: Amyris, 5885 Hollis Street, Suite 100, Emeryville, California 94608, USA.
authors.forename: Peter
authors.initials: P
authors.lastname: Jackson
authors.affiliation: Amyris, 5885 Hollis Street, Suite 100, Emeryville, California 94608, USA.
authors.forename: Robert
authors.initials: R
authors.lastname: Mans
authors.affiliation: Amyris, 5885 Hollis Street, Suite 100, Emeryville, California 94608, USA.
authors.forename: Darren
authors.initials: D
authors.lastname: Platt
authors.affiliation: Amyris, 5885 Hollis Street, Suite 100, Emeryville, California 94608, USA.
authors.forename: Christopher D
authors.initials: CD
authors.lastname: Reeves
authors.affiliation: Amyris, 5885 Hollis Street, Suite 100, Emeryville, California 94608, USA.
authors.forename: Poonam R
authors.initials: PR
authors.lastname: Saija
authors.affiliation: Amyris, 5885 Hollis Street, Suite 100, Emeryville, California 94608, USA.
authors.forename: Gale
authors.initials: G
authors.lastname: Wichmann
authors.affiliation: Amyris, 5885 Hollis Street, Suite 100, Emeryville, California 94608, USA.
authors.forename: Victor F
authors.initials: VF
authors.lastname: Holmes
authors.affiliation: Amyris, 5885 Hollis Street, Suite 100, Emeryville, California 94608, USA.
authors.forename: Kirsten
authors.initials: K
authors.lastname: Benjamin
authors.affiliation: Amyris, 5885 Hollis Street, Suite 100, Emeryville, California 94608, USA.
authors.forename: Paul W
authors.initials: PW
authors.lastname: Hill
authors.affiliation: Amyris, 5885 Hollis Street, Suite 100, Emeryville, California 94608, USA.
authors.forename: Timothy S
authors.initials: TS
authors.lastname: Gardner
authors.affiliation: Amyris, 5885 Hollis Street, Suite 100, Emeryville, California 94608, USA.
authors.forename: Annie E
authors.initials: AE
authors.lastname: Tsong
mesh_terms: D000105:Acetyl Coenzyme A / Q000096:biosynthesis / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D019149:Bioreactors; D053898:Biosynthetic Pathways; D050260:Carbohydrate Metabolism; D002244:Carbon / Q000378:metabolism*; D002245:Carbon Dioxide / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D005285:Fermentation; D060847:Metabolic Engineering; D010084:Oxidation-Reduction; D010100:Oxygen / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000378:metabolism*; D012717:Sesquiterpenes / Q000378:metabolism; D013729:Terpenes / Q000378:metabolism*
chemical_list: D012717:Sesquiterpenes; D013729:Terpenes; D002245:Carbon Dioxide; C062671:beta-farnesene; D000105:Acetyl Coenzyme A; D002244:Carbon; D000255:Adenosine Triphosphate; D010100:Oxygen