title: Evolution reveals a glutathione-dependent mechanism of 3-hydroxypropionic acid tolerance.
abstract: Biologically produced 3-hydroxypropionic acid (3 HP) is a potential source for sustainable acrylates and can also find direct use as monomer in the production of biodegradable polymers. For industrial-scale production there is a need for robust cell factories tolerant to high concentration of 3 HP, preferably at low pH. Through adaptive laboratory evolution we selected S. cerevisiae strains with improved tolerance to 3 HP at pH 3.5. Genome sequencing followed by functional analysis identified the causal mutation in SFA1 gene encoding S-(hydroxymethyl)glutathione dehydrogenase. Based on our findings, we propose that 3 HP toxicity is mediated by 3-hydroxypropionic aldehyde (reuterin) and that glutathione-dependent reactions are used for reuterin detoxification. The identified molecular response to 3 HP and reuterin may well be a general mechanism for handling resistance to organic acid and aldehydes by living cells.
authors.affiliation: The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kogle Allé 6, DK-2970 Hørsholm, Denmark.
authors.forename: Kanchana R
authors.initials: KR
authors.lastname: Kildegaard
authors.affiliation: Science for Life Laboratory, KTH Royal Institution of Technology, Box 1031, SE-171 21 Solna, Sweden.
authors.forename: Björn M
authors.initials: BM
authors.lastname: Hallström
authors.affiliation: The Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Blegdamsvej 3b, DK-2200 Copenhagen , Denmark.
authors.forename: Thomas H
authors.initials: TH
authors.lastname: Blicher
authors.affiliation: The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kogle Allé 6, DK-2970 Hørsholm, Denmark.
authors.forename: Nikolaus
authors.initials: N
authors.lastname: Sonnenschein
authors.affiliation: The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kogle Allé 6, DK-2970 Hørsholm, Denmark.
authors.forename: Niels B
authors.initials: NB
authors.lastname: Jensen
authors.affiliation: The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kogle Allé 6, DK-2970 Hørsholm, Denmark.
authors.forename: Svetlana
authors.initials: S
authors.lastname: Sherstyk
authors.affiliation: The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kogle Allé 6, DK-2970 Hørsholm, Denmark.
authors.forename: Scott J
authors.initials: SJ
authors.lastname: Harrison
authors.affiliation: The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kogle Allé 6, DK-2970 Hørsholm, Denmark.
authors.forename: Jérôme
authors.initials: J
authors.lastname: Maury
authors.affiliation: The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kogle Allé 6, DK-2970 Hørsholm, Denmark.
authors.forename: Markus J
authors.initials: MJ
authors.lastname: Herrgård
authors.affiliation: The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kogle Allé 6, DK-2970 Hørsholm, Denmark.
authors.forename: Agnieszka S
authors.initials: AS
authors.lastname: Juncker
authors.affiliation: The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kogle Allé 6, DK-2970 Hørsholm, Denmark.
authors.forename: Jochen
authors.initials: J
authors.lastname: Forster
authors.affiliation: The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kogle Allé 6, DK-2970 Hørsholm, Denmark; Department of Chemical and Biological Engineering, Chalmers University of Technology, Kemivägen 10, SE-412 96 Göteborg, Sweden.
authors.forename: Jens
authors.initials: J
authors.lastname: Nielsen
authors.affiliation: The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kogle Allé 6, DK-2970 Hørsholm, Denmark. Electronic address: irbo@biosustain.dtu.dk.
authors.forename: Irina
authors.initials: I
authors.lastname: Borodina
mesh_terms: D049109:Cell Proliferation / Q000187:drug effects / Q000235:genetics; D019020:Directed Molecular Evolution / Q000379:methods*; D004305:Dose-Response Relationship, Drug; D004361:Drug Tolerance / Q000235:genetics; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics*; D024861:Genetic Enhancement / Q000379:methods*; D005978:Glutathione / Q000235:genetics / Q000378:metabolism*; D019344:Lactic Acid / Q000008:administration & dosage / Q000031:analogs & derivatives*; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics*
chemical_list: D019344:Lactic Acid; C031601:hydracrylic acid; D005978:Glutathione
keywords: 3-hydroxypropionic acid; 3-hydroxypropionic aldehyde (reuterin); Adaptive laboratory evolution; Saccharomyces cerevisiae; Tolerance