Self-Cloning Yeast Strains Containing Novel FAS2 Mutations Produce a Higher Amount of Ethyl Caproate in Japanese Sake
Biosci. Biotechnol. Biochem., 68 (1), 206–214, 2004 Self-cloning Yeast Strains Containing Novel FAS2 Mutations Produce a Higher Amount of Ethyl Caproate in Japanese Sake Kazuo ARITOMI,1 Isao HIROSAWA,2 Hisashi HOSHIDA,2 Mikio SHIIGI,1 y Yoshinori NISHIZAWA,2 Susumu KASHIWAGI,1 and Rinji AKADA2; 1Yamaguchi Prefectural Industrial Technology Institute, Asutopia, Ube 755-0151, Japan 2Department of Applied Chemistry and Chemical Engineering, Faculty of Engineering, Yamaguchi University, Tokiwadai, Ube 755-8611, Japan Received August 25, 2003; Accepted September 27, 2003 Point mutation of Gly1250Ser (1250S) of the yeast inhibitor of fatty acid synthesis,4,5) has been used to fatty acid synthase gene FAS2 confers cerulenin resist- isolate yeast mutants that produced higher amounts of ance. This mutation also results in a higher production ethyl caproate, an apple-like flavor component in sake.3) of the apple-like flavor component ethyl caproate in The cerulenin resistance and the higher production of Japanese sake. We mutated the 1250th codon by in vitro ethyl caproate were conferred by a mutation from site-directed mutagenesis to encode Ala (1250A) or Cys glycine to serine at the 1250th codon (1257th in a (1250C) and examined cerulenin resistance and ethyl different strain) of the fatty acid synthase gene FAS2.6–9) caproate production. The mutated FAS2 genes were This dominant FAS2 mutation was thought to decrease inserted into a binary plasmid vector containing a drug- the activity of the fatty acid synthase complex, which is resistance marker and a counter-selectable marker, a hexamer ( 6 6) of and subunits encoded by FAS2 GALp-GIN11M86.
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