Dimethyl trisulfide (DMTS) is one of the components responsible for the unpalatable aroma of stale Japanese sake, called "hineka". Recently, a precursor compound of DMTS, 1,2-dihydroxy-5-(methylsulfinyl)pentan-3-one (DMTS-P1), was identified. It was speculated that the yeast methionine salvage pathway (MTA cycle) might participate in the formation of DMTS-P1, because the chemical structure of DMTS-P1 was similar to one of the intermediate compounds of that pathway. Here, we carried out sake brewing tests using laboratory yeast strains with disrupted MTA cycle genes and found that DMTS-P1 was hardly produced by ?meu1, ?mri1, and ?mde1 strains. Furthermore, the DMTS producing potential (production of DMTS during storage of sake) decreased in sake made with ?mri1 and ?mde1. We constructed sake yeast strains with a disrupted MRI1 or MDE1 gene and confirmed a decline in the DMTS-P1 content and DMTS producing potential of sake made with these disruptants. The results of sake brewing tests using MTA cycle disruptants suggested that SPE2 is responsible for the production of DMTS precursors other than DMTS-P1: although the DMTS-P1 content was higher in ?spe2 sake than in ?mri1 or ?mde1 sake, the DMTS producing potential of ?spe2 sake was as low as that of ?mri1 or ?mde1 sake. Sake brewing tests using BY4743 ?spe2 ?mri1 double disruptants revealed that the DMTS producing potential was further decreased as compared with the ?spe2 or ?mri1 single disruptant. These results suggest that MRI1, MDE1, and SPE2 are promising targets for breeding yeast to suppress the formation of DMTS during storage of sake.
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