A simple genotyping method for RIM15 gene polymorphisms in sake yeast strains

Takashi Kuribayashi; Nanako Asahi; Hina Satone; Jumpei Tanaka; Masamichi Sugawara; Keigo Sato; Yoshihito Nabekura; Toshio Joh; Toshio Aoki

doi:10.51094/jxiv.868

##article.authors##

Takashi Kuribayashi Niigata Prefectural Sake Research Institute / Faculty of Food Industry, Niigata Agro-Food University https://researchmap.jp/takashi-kuribayashi
Nanako Asahi Faculty of Food Industry, Niigata Agro-Food University
Hina Satone Faculty of Food Industry, Niigata Agro-Food University
Jumpei Tanaka Niigata Prefectural Sake Research Institute
Masamichi Sugawara Niigata Prefectural Sake Research Institute
Keigo Sato Niigata Prefectural Sake Research Institute
Yoshihito Nabekura Niigata Prefectural Sake Research Institute
Toshio Joh Faculty of Agriculture, Niigata University
Toshio Aoki Niigata Prefectural Sake Research Institute

DOI:

https://doi.org/10.51094/jxiv.868

キーワード:

polymerase chain reaction-restriction fragment length polymorphism、 RIM15、 Saccharomyces cerevisiae

抄録

In the process of brewing sake, Saccharomyces cerevisiae not only synthesizes numerous flavor compounds but also produces alcohol, which enhances fermentation efficiency. A previous study identified a sake yeast mutant with a single-nucleotide insertion in the RIM15 protein kinase gene (RIM15 with an insertion of adenine at position 5067: RIM15ins5067A). In this study, we developed primer sets (P1/P2) with engineered mismatches that introduce restriction sites for the restriction enzyme CpsCI. This allowed us to create a simple and rapid polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay to identify Kyokai no. 7 (K7) group sake yeast strains carrying the RIM15ins5067A mutation. We validated the PCR-RFLP method in detecting the RIM15ins5067A mutation in the K7 group of yeast strains. This approach provides a straightforward, effective, and valuable tool for distinguishing specific yeasts used in sake production.

利益相反に関する開示

The authors declare no conflict of interest. All the experiments undertaken in this study complied with the current laws of the country in which they were performed.

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