Identification of a mutation causing a defective spindle assembly checkpoint in high ethyl caproate-producing sake yeast strain K1801

• Published in: Bioscience, biotechnology, and biochemistry Vol. 80; no. 8; pp. 1657 - 1662
• Main Authors: Goshima, Tetsuya, Nakamura, Ryo, Kume, Kazunori, Okada, Hiroki, Ichikawa, Eri, Tamura, Hiroyasu, Hasuda, Hirokazu, Inahashi, Masaaki, Okazaki, Naoto, Akao, Takeshi, Shimoi, Hitoshi, Mizunuma, Masaki, Ohya, Yoshikazu, Hirata, Dai
• Format: Journal Article
• Language: English
• Published: England Taylor & Francis 02.08.2016
• Subjects: Alcoholic Beverages; Amino Acid Sequence; Base Sequence; Caproates - metabolism; Cell Cycle Proteins - genetics; Cell Cycle Proteins - metabolism; cerulenin-resistant; checkpoint; Ethanol - metabolism; ethyl caproate; Fermentation; Food Technology; Gene Expression; Humans; Japan; M Phase Cell Cycle Checkpoints; morphological variation; Mutation; Odorants; Oryza - chemistry; Polymorphism, Single Nucleotide; Protein Phosphatase 2 - genetics; Protein Phosphatase 2 - metabolism; Saccharomyces cerevisiae - genetics; Saccharomyces cerevisiae - metabolism; Saccharomyces cerevisiae Proteins - genetics; Saccharomyces cerevisiae Proteins - metabolism; sake yeast; Selection, Genetic; checkpoint; ethyl caproate; cerulenin-resistant; sake yeast; morphological variation
• ISSN: 0916-8451; 1347-6947
• DOI: 10.1080/09168451.2016.1184963

In high-quality sake brewing, the cerulenin-resistant sake yeast K1801 with high ethyl caproate-producing ability has been used widely; however, K1801 has a defective spindle assembly checkpoint (SAC). To identify the mutation causing this defect, we first searched for sake yeasts with a SAC-defect like K1801 and found that K13 had such a defect. Then, we searched for a common SNP in only K1801 and K13 by examining 15 checkpoint-related genes in 23 sake yeasts, and found 1 mutation, R48P of Cdc55, the PP2A regulatory B subunit that is important for the SAC. Furthermore, we confirmed that the Cdc55-R48P mutation was responsible for the SAC-defect in K1801 by molecular genetic analyses. Morphological analysis indicated that this mutation caused a high cell morphological variation. But this mutation did not affect the excellent brewing properties of K1801. Thus, this mutation is a target for breeding of a new risk-free K1801 with normal checkpoint integrity. K1801 (triploid) having 2 copies of R48P-type cdc55 and 1 copy of R48-type cdc55 showed a defective spindle assembly checkpoint (SAC). Cdc55-R48P is semidominant mutation in the SAC function.