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When Bacillus subtilis NB-NUC1 associated with Cheonggukjang fermentation was aerobically grown in a synthetic medium containing 1 to 2% glycine (w/v), cell growth was inhibited in a dose-dependent manner. Subsequently, different concentrations of glycine (0, 1, and 2%) were used in Cheonggukjang fermentation for 96 h at 40 °C. Supplementation of 1% glycine increased extracellular γ-glutamyl transpeptidase (γ-GTPase), responsible for the production of viscous substance. Based on correlation studies, we conclude that the production of viscous substance is correlated with viscous extension (r = 0.867), extracellular proteins contents (r = 0.821), and γ-GTPase activity (r = 0.807). The molecular weight of the viscous substance obtained during Cheonggukjang fermentation by B. subtilis NB-NUC1 was also affected by glycine supplementation. Our results demonstrate that glycine supplementation before solid-state fermentation may increase the mass production of mucilage in food industry.