Conference paper
Evolution Reveals A Glutathione-dependent Mechanism Of 3-hydroxypropionic Acid Detoxification
Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark1
Research Groups, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark2
KTH Royal Institute of Technology3
University of Copenhagen4
iLoop, Translational Management, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark5
CFB - Core Flow, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark6
Bacterial Cell Factories, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark7
Metagenomics, Center for Biological Sequence Analysis, Department of Systems Biology, Technical University of Denmark8
Fungal Cell Factories, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark9
Biologically produced 3-hydroxypropionic acid (3HP) 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, high titer, rate and yield are essential; thus there is a need for robust cell factories tolerant to high concentration of 3HP, preferably at low pH.
Through adaptive laboratory evolution we selected S. cerevisiae strains with improved tolerance to 3HP at pH 3.5. Genome sequencing of three independent clones identified single-nucleotide changes in the SFA1 gene encoding S-(hydroxymethyl)glutathione dehydrogenase. Introduction of the mutated SFA1 alleles or overexpression of any of the SFA1 alleles in a sfa16 strain enabled growth in the presence of above 40 g/L 3HP.
We further found that aldehyde dehydrogenase (ALD6), S-formylglutathione hydrolase (YJL068C) and glutathione play a role in 3HP detoxicification. Addition of glutathione relieved growth inhibition by 3HP for several yeast species and for E. coli; but glutathione could not enable growth of a S. cerevisiae sfa16 strain.
Based on our findings we propose a 3-hydroxypropionic aldehyde-mediated mechanism underlying 3HP toxicity as well as a glutathione-dependent route for detoxification of 3-hydroxypropionic aldehyde (reuterin). The identified molecular response to 3HP and reuterin may well be a general mechanism for handling resistance to organic acids and aldehydes by living cells
Language: | English |
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Year: | 2015 |
Proceedings: | Metabolic Engineering X |
Types: | Conference paper |
ORCIDs: | Kildegaard, Kanchana Rueksomtawin , Sonnenschein, Nikolaus , Maury, Jerome , Herrgard, Markus , Förster, Jochen and Borodina, Irina |