A simple genotyping method for RIM15 gene polymorphisms in sake yeast strains
Takashi Kuribayashi1,2, Nanako Asahi2, Hina Satone2
1a Niigata Prefectural Sake Research Institute.
Mycoscience
|April 13, 2026
Summary
A new PCR-RFLP method quickly identifies sake yeast strains with a specific RIM15 gene mutation. This technique aids in distinguishing yeasts crucial for producing distinct sake flavors and alcohol content.
Area of Science:
- Microbiology
- Biotechnology
- Food Science
Background:
- Sake brewing relies on Saccharomyces cerevisiae for alcohol and flavor compound production.
- A specific mutation, RIM15ins5067A, in the RIM15 gene creates a unique sake yeast strain.
- Accurate identification of sake yeast strains is vital for consistent sake production.
Purpose of the Study:
- To develop a rapid and simple method for identifying sake yeast strains with the RIM15ins5067A mutation.
- To enable precise differentiation of Kyokai no. 7 (K7) group sake yeasts.
- To provide a valuable tool for quality control in sake production.
Main Methods:
- Designed primer sets (P1/P2) with engineered mismatches to introduce CspCI restriction sites.
- Developed a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay.
- Validated the PCR-RFLP assay for detecting the RIM15ins5067A mutation in K7 yeast strains.
Main Results:
- Successfully created a simple and rapid PCR-RFLP assay.
- The assay effectively identified the RIM15ins5067A mutation in K7 sake yeast strains.
- Demonstrated the reliability of the developed method for strain identification.
Conclusions:
- The developed PCR-RFLP assay is a straightforward and effective tool.
- This method facilitates the distinction of specific sake yeast strains.
- The assay supports quality control and research in sake production.
