Applied Biochemistry and Biotechnology

, Volume 189, Issue 3, pp 810–821 | Cite as

Improved Stress Tolerance of Saccharomyces cerevisiae by CRISPR-Cas-Mediated Genome Evolution

  • Ryosuke Mitsui
  • Ryosuke YamadaEmail author
  • Hiroyasu Ogino


In bioprocesses, a microorganism with high tolerance to various stresses would be advantageous for efficient bio-based chemical production. Yeast Saccharomyces cerevisiae has long been used in the food industry because of its safety and convenience, and genetically engineered S. cerevisiae strains have been constructed and used for the production of various bio-based chemicals. In this study, we developed a novel genome shuffling method for S. cerevisiae using CRISPR-Cas. By using this, the thermotolerant mutant strain T8-292, which can grow well at 39 °C, was successfully created. The strain also showed higher cell viability in low pH and high ethanol concentration. In addition, the differences in genome structure between mutant and parent strains were suggested by random amplified polymorphic DNA PCR method. Our genome shuffling method could be a promising strategy for improvement of various stress tolerance in S. cerevisiae.


CRISPR-Cas δ sequence DNA repair Genome shuffling Saccharomyces cerevisiae Stress tolerance 



This work was partly supported by the Japan Society for the Promotion of Science KAKENHI (grant number JP18K14069) and KAKENHI Specific Support Operation of Osaka Prefecture University to RY.

Supplementary material

12010_2019_3040_MOESM1_ESM.docx (112 kb)
ESM 1 (DOCX 111 kb)


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© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Chemical EngineeringOsaka Prefecture UniversityOsakaJapan

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