We collected 92 isolates belonging to the genus Bacillus from the sake brewing process at Shiraki Tsunesuke Sake Brewery in Gifu, Japan to determine whether there is strain specificity at individual sake breweries. After distributing the isolates into seven groups, we observed that at least two groups (68 isolates) were kuratsuki bacteria at Shiraki Tsunesuke Sake Brewery. The kuratsuki Bacillus isolates were collected from different samples at the early and late stages of sake brewing in 2021 and 2019, respectively. These results showed that kuratsuki Bacillus entered the sake brewing process at this location. These kuratsuki Bacillus isolates had a high ethanol tolerance. Our previous paper showed the existence of kuratsuki Kocuria at Narimasa Sake Brewery in Toyama, Japan, but this study demonstrated that it is not found at Shiraki Tsunesuke Sake Brewery. Therefore, each sake brewery has specific kuratsuki bacterial strains, which are isolated with high frequency and contribute a specific flavor or taste to each sake brewery.