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Rapid and efficient galactose fermentation by engineered Saccharomyces cerevisiae.

Josh Quarterman1, Jeffrey M Skerker2, Xueyang Feng3

  • 1Department of Food Science and Human Nutrition, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA; Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA.

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Summary

Researchers engineered yeast to ferment galactose anaerobically, enhancing ethanol production. Deleting COX9 and mutating Gal80p enabled efficient fermentation, creating superior industrial strains.

Keywords:
EthanolEvolutionary engineeringGalactoseGenome sequencingSaccharomyces cerevisiaeSystems biology

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Area of Science:

  • Biotechnology and metabolic engineering
  • Industrial microbiology
  • Yeast fermentation

Background:

  • Saccharomyces cerevisiae typically requires respiration for galactose metabolism, limiting anaerobic applications.
  • Current galactose metabolism is inefficient for fuel and chemical production due to CO2 byproduct formation.
  • Engineering yeast for anaerobic galactose fermentation is crucial for sustainable bioprocessing.

Purpose of the Study:

  • To develop a method for enabling Saccharomyces cerevisiae to utilize galactose via fermentation under anaerobic conditions.
  • To enhance ethanol yield and productivity from galactose in industrial yeast strains.
  • To investigate the genetic and metabolic basis for anaerobic galactose fermentation.

Main Methods:

  • Deletion of the COX9 gene to impair respiration in Saccharomyces cerevisiae.
  • Serial sub-culturing on galactose to select for evolved fermentative strains.
  • (13)C-metabolic flux analysis and genome sequencing to characterize metabolic and genetic changes.
  • Construction of double deletion mutants (Δcox9Δgal80) for synergistic analysis.

Main Results:

  • An evolved strain (JQ-G1) demonstrated efficient galactose fermentation with a 94% increase in ethanol yield and 6.9-fold higher specific productivity.
  • Metabolic flux analysis revealed reduced TCA cycle activity and redirected flux towards fermentation in the evolved strain.
  • A loss-of-function mutation in Gal80p (Glu348*) synergized with COX9 deletion, significantly improving fermentation rates and yields.

Conclusions:

  • The metabolic 'death valley' (impaired galactose utilization in Δcox9 mutants) is a critical intermediate for evolving fermentative capabilities.
  • Adaptive evolution combined with targeted gene deletions offers a powerful strategy for generating yeast strains with enhanced anaerobic fermentation phenotypes.
  • This study presents a promising approach for optimizing yeast for sustainable biofuel and biochemical production from galactose.