title: Improved Acetic Acid Resistance in Saccharomyces cerevisiae by Overexpression of the WHI2 Gene Identified through Inverse Metabolic Engineering.
abstract: Development of acetic acid-resistant Saccharomyces cerevisiae is important for economically viable production of biofuels from lignocellulosic biomass, but the goal remains a critical challenge due to limited information on effective genetic perturbation targets for improving acetic acid resistance in the yeast. This study employed a genomic-library-based inverse metabolic engineering approach to successfully identify a novel gene target, WHI2 (encoding a cytoplasmatic globular scaffold protein), which elicited improved acetic acid resistance in S. cerevisiae. Overexpression of WHI2 significantly improved glucose and/or xylose fermentation under acetic acid stress in engineered yeast. The WHI2-overexpressing strain had 5-times-higher specific ethanol productivity than the control in glucose fermentation with acetic acid. Analysis of the expression of WHI2 gene products (including protein and transcript) determined that acetic acid induced endogenous expression of Whi2 in S. cerevisiae. Meanwhile, the whi2Δ mutant strain had substantially higher susceptibility to acetic acid than the wild type, suggesting the important role of Whi2 in the acetic acid response in S. cerevisiae. Additionally, overexpression of WHI2 and of a cognate phosphatase gene, PSR1, had a synergistic effect in improving acetic acid resistance, suggesting that Whi2 might function in combination with Psr1 to elicit the acetic acid resistance mechanism. These results improve our understanding of the yeast response to acetic acid stress and provide a new strategy to breed acetic acid-resistant yeast strains for renewable biofuel production.
authors.affiliation: Department of Civil and Environmental Engineering and Earth Sciences, University of Notre Dame, Notre Dame, Indiana, USA.
authors.forename: Yingying
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Department of Civil and Environmental Engineering, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.
authors.forename: Lisa
authors.initials: L
authors.lastname: Stabryla
authors.affiliation: Department of Civil and Environmental Engineering and Earth Sciences, University of Notre Dame, Notre Dame, Indiana, USA nwei@nd.edu.|Department of Civil and Environmental Engineering, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.
authors.forename: Na
authors.initials: N
authors.lastname: Wei
mesh_terms: D019342:Acetic Acid / Q000378:metabolism*; D000431:Ethanol / Q000378:metabolism; D005285:Fermentation; D015870:Gene Expression; D005947:Glucose / Q000378:metabolism; D060847:Metabolic Engineering; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics* / Q000378:metabolism; D014994:Xylose / Q000378:metabolism
chemical_list: D029701:Saccharomyces cerevisiae Proteins; C470937:Whi2 protein, S cerevisiae; D000431:Ethanol; D014994:Xylose; D005947:Glucose; D019342:Acetic Acid
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