Self-cloning Yeast Strains Containing Novel<i>FAS2</i>Mutations Produce a Higher Amount of Ethyl Caproate in Japanese Sake

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  • ARITOMI Kazuo
    Yamaguchi Prefectural Industrial Technology Institute
  • HIROSAWA Isao
    Department of Applied Chemistry and Chemical Engineering, Faculty of Engineering, Yamaguchi University
  • HOSHIDA Hisashi
    Department of Applied Chemistry and Chemical Engineering, Faculty of Engineering, Yamaguchi University
  • SHIIGI Mikio
    Yamaguchi Prefectural Industrial Technology Institute
  • NISHIZAWA Yoshinori
    Department of Applied Chemistry and Chemical Engineering, Faculty of Engineering, Yamaguchi University
  • KASHIWAGI Susumu
    Yamaguchi Prefectural Industrial Technology Institute
  • AKADA Rinji
    Department of Applied Chemistry and Chemical Engineering, Faculty of Engineering, Yamaguchi University

書誌事項

タイトル別名
  • Self-cloning Yeast Strains Containing Novel FAS2 Mutations Produce a Higher Amount of Ethyl Caproate in Japanese Sake
  • Self-cloning yeast strain containing novel FAS2 mutations produce a higher amount of ethyl caproate in Japanese sake
公開日
2004
資源種別
journal article
DOI
  • 10.1271/bbb.68.206
公開者
公益社団法人 日本農芸化学会

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説明

Point mutation of Gly1250Ser (1250S) of the yeast fatty acid synthase gene FAS2 confers cerulenin resistance. This mutation also results in a higher production of the apple-like flavor component ethyl caproate in Japanese sake. We mutated the 1250th codon by in vitro site-directed mutagenesis to encode Ala (1250A) or Cys (1250C) and examined cerulenin resistance and ethyl caproate production. The mutated FAS2 genes were inserted into a binary plasmid vector containing a drug-resistance marker and a counter-selectable marker, GALp-GIN11M86. The plasmids were integrated into the wild-type FAS2 locus of a sake yeast strain, and the loss of the plasmid sequences from the integrants was done by growth on galactose plates, which is permissive for loss of GALp-GIN11M86. These counter-selected strains contained either the wild type or the mutated FAS2 allele but not the plasmid sequences, from which FAS2 mutant strains were selected by allele-specific PCR. The FAS2-1250C mutant produced a higher amount of ethyl caproate in sake than FAS2-1250S, while FAS2-1250A produced an ethyl caproate level intermediate between FAS2-1250S and the parental Kyokai no. 7 strain. Interestingly, these mutants only showed detectable cerulenin resistance. These ‘self-cloning’ yeast strains should be acceptable to the public because they can improve sake quality without the presence of extraneous DNA sequences.

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