Abstract

Sake is a traditional Japanese alcoholic beverage prepared by multiple parallel fermentation of rice. The fermentation process of “yamahai-ginjo-shikomi” sake is mainly performed by three microbes, Aspergillus oryzae, Saccharomyces cerevisiae, and Lactobacilli; the levels of various metabolites fluctuate during the fermentation of sake. For evaluation of the fermentation process, we monitored the concentration of moderate-sized molecules (m/z: 200–1000) dynamically changed during the fermentation process of “yamahai-ginjo-shikomi” Japanese sake. This analysis revealed that six compounds were the main factors with characteristic differences in the fermentation process. Among the six compounds, four were leucine- or isoleucine-containing peptides and the remaining two were predicted to be small molecules. Quantification of these compounds revealed that their quantities changed during the month of fermentation process. Our metabolomic approach revealed the dynamic changes observed in moderate-sized molecules during the fermentation process of sake, and the factors found in this analysis will be candidate molecules that indicate the progress of “yamahai-ginjo-shikomi” sake fermentation.

Highlights

  • Sake is a traditional Japanese rice wine prepared by multistep fermentation of rice, called multiple parallel fermentation, in which conversions of rice starch to sugar by Aspergillus oryzae and sugar to alcohol by sake yeast (Saccharomyces cerevisiae) spontaneously occur[1]

  • To profile the metabolite changes of the fermentation processes, we prepared the sake mash samples taken during sake fermentation process at 6, 11, 16, 21, 26, and 34 days after “tome-shikomi” for sake_1 sake mash sample, and at 1, 5, 11, 19, 26, and 34 days after “tome-shikomi” for sake_2 sake mash samples

  • We performed the time-course metabolic profiling of “yamahai-ginjo-shikomi” Japanese sake and first identified factors that indicate the progress of the fermentation process

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Summary

Introduction

Sake is a traditional Japanese rice wine prepared by multistep fermentation of rice, called multiple parallel fermentation, in which conversions of rice starch to sugar by Aspergillus oryzae and sugar to alcohol by sake yeast (Saccharomyces cerevisiae) spontaneously occur[1]. The process of brewing sake begins with polishing of rice grains to remove bran-containing proteins, lipids, and minerals that could impair the aroma and flavor of sake. The sake using rice polished to at least 60% of the original grain was called as “ginjo” sake. A fermentation starter is prepared using steamed rice, malted rice, and sake yeast.

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