Mutagenesis, breeding, and characterization of sake yeast strains with low production of dimethyl trisulfide precursor
Section snippets
Yeast strains and media
The sake yeast Saccharomyces cerevisiae strain Kyokai no. 701 (K701) was provided by the Brewing Society of Japan. Two additional haploid strains derived from K701 owned by the Nihonsakari Co., Ltd. (Nishinomiya, Japan), 9-1 (MATα) and #4 (MATa), were also used. The methionine-auxotrophic strain (MO-001, MATα) was obtained from 9-1 strain. MDR01 was produced by the disruption of the MRI1 gene of MO-001 strain.
YPD medium (1% yeast extract, 2% bacto-peptone, and 2% glucose) was used for yeast
Establishment of a screening method for MTA non-utilizing strains
The methionine-auxotrophic strain MO-001 was generated from the wild-type strain 9-1 by EMS mutation treatment, and was selected from strains that did not grow in the minimal culture medium, but grew in minimal culture medium containing methionine, and formed black or brown colonies due to the generation of lead sulfide in MLA medium, using the replica method. The medium conditions were examined using the MRI1 gene disrupted strain MDR01 obtained from MO-001 strain. First, acquisition was
Discussion
In this study, the MTA method was established as a screening method for MTA non-utilizing strains targeting genes involved in the production of DMTS-P1, a precursor of DMTS and the main component of hineka. The mutants were screened using the MTA method to obtain candidate strains. These candidate strains were confirmed to have mutations in the MDE1 or MRI1 gene, and the DMTS-P1 production ability was low, as expected. However, the fermentation abilities of the strains were poor, and therefore,
Acknowledgments
We wish to thank Dr. T. Goshima for technical suggestions on FACS analysis.
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Present address: Faculty of Food and Agricultural Sciences, Fukushima University, 1 Kanayagawa, Fukushima 960-1296, Japan.