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The Green Monster Process for the Generation of Yeast Strains Carrying Multiple Gene Deletions
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Polyploid engineering by increasing mutant gene dosage in yeasts.

Nobuo Fukuda1,2, Shinya Honda2, Maki Fujiwara3

  • 1Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Osaka, Japan.

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Yeast polyploids with adjusted gene copy numbers can regulate ethanol fermentation. Increasing mutant gene dosage in tetraploid yeast strains enhances ethyl caproate production, a key flavor compound.

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Area of Science:

  • Microbiology
  • Biotechnology
  • Industrial Fermentation

Background:

  • Saccharomyces cerevisiae is a key organism for ethanol production, with diploid strains showing superior industrial performance.
  • Whole-genome duplications and allelic variations influence microbial adaptation and product composition during fermentation.
  • Fine-tuning fermentation products is challenging, despite various yeast strains available.

Purpose of the Study:

  • To investigate the use of polyploid yeast with varying gene dosage for regulating ethanol fermentation.
  • To explore the impact of mutant gene dosage on the production of ethyl caproate, a sake flavoring agent.

Main Methods:

  • Generated and evaluated yeast strains with varying gene dosage using FAS2 as a model.
  • Utilized loss of heterozygosity in diploid and tetraploid strains to increase mutant gene dosage.
  • Assessed ethyl caproate productivity in relation to mutant gene dosage.

Main Results:

  • Successfully increased mutant gene dosage in diploid and tetraploid yeast strains.
  • Demonstrated a gradual increase in ethyl caproate productivity with increasing mutant gene dosage in tetraploid strains.

Conclusions:

  • Polyploid yeast with controlled gene dosage offers a method for regulating fermentation products.
  • This approach shows potential for developing yeast strains with enhanced production of specific compounds like ethyl caproate.
  • Growth-based screening can be applied for yeast strain development using this gene dosage strategy.