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Anticancer effects of chungkukjang (a Korean short-term fermented soy paste) were studied in human gastric adenocarcinoma cells, and Bacillus strains from chungkukjang were isolated and identified. Before the experiments, six different chungkukjang products (K-, M-, Mn-, O-, Os-, and H-chungkukjangs) were purchased from a folk village in the Sunchang region, Jeonbuk, Republic of Korea. Based on sensory evaluation tests and general chemical and quality studies, K-, H-, and M-chungkukjangs were selected for the experiments. All chungkukjang samples exhibited in vitro anticancer activities; however, K-chungkukjang revealed the highest anticancer activity in the previous studies. In this experiment, K-chungkukjang again showed the highest anticancer effect in the AGS cells. At the concentration of 1mg/mL, K-chungkukjang (87%) showed the highest growth inhibitory effect, followed by H-chungkukjang (85%) and MC-chungkukjang (69%) (P<.05). K-chungkukjang induced apoptosis as determined by 4,6-diamidino-2-phenylindole staining and exhibited increased bax and decreased bcl-2 mRNA expression. Three representative Bacillus strains from K-chungkukjang were isolated and identified by recA gene sequencing as Bacillus cereus, Bacillus amyloliquefaciens, and Bacillus subtilis. Identifying B. cereus in the chungkukjang means that when chungkukjang is prepared by the traditional method, B. cereus, which is a common cause of foodborne disease, can grow during the natural fermentation process. All B. cereus strains, of course, are not pathogens, but its presence causes food safety concerns. Therefore, using a starter culture is safer than the traditional natural fermentation for the industrialization of chungkukjang in Korea.