Self-cloning Yeast Strains Containing Novel FAS2 Mutations Produce a Higher Amount of Ethyl Caproate in Japanese Sake
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概要
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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.
- 社団法人 日本農芸化学会の論文
- 2004-01-23
著者
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HOSHIDA Hisashi
Department of Applied Molecular Bioscience, Yamaguchi University Graduate School of Medicine
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AKADA Rinji
Department of Applied Molecular Bioscience, Yamaguchi University Graduate School of Medicine
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Akada Rinji
Department Of Applied Chemistry And Chemical Engineering Faculty Of Engineering Yamaguchi University
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NISHIZAWA YOSHINORI
Department of Fermentation Tachnology, Faculty of Engineering, Hiroshima University
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Nishizawa Y
Department Of Fermentation Technology Faculty Of Engineering Hiroshima University
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Nishizawa Y
Yamaguchi Univ. Ube Jpn
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Nishizawa Yoshinori
Department Of Applied Chemistry And Chemical Engineering Faculty Of Engineering Yamaguchi University
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Nishizawa Yoshinori
Department Of Fermentation Technology Faculty Of Engineering Hiroshima University
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Hoshida Hisashi
Department Of Applied Chemistry And Chemical Engineering Faculty Of Engineering Yamaguchi University
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ARITOMI Kazuo
Yamaguchi Prefectural Industrial Technology Institute
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HIROSAWA Isao
Department of Applied Chemistry and Chemical Engineering, Faculty of Engineering, Yamaguchi Universi
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SHIIGI Mikio
Yamaguchi Prefectural Industrial Technology Institute
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KASHIWAGI Susumu
Yamaguchi Prefectural Industrial Technology Institute
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Aritomi Kazuo
Yamaguchi Prefectural Industrial Technol. Inst.
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Hirosawa Isao
Department Of Applied Chemistry And Chemical Engineering Faculty Of Engineering Yamaguchi University
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