Diabetes is a serious disease, the destruction was terrible, it causes disability for diabetic patients, especially untreated or inadequately treated patients. Moranoline (1-Deoxynojirimycin, DNJ) as a potential aglucosidase inhibitor produced by Bacillus strain used for diabetes type 2 treatment. Objective: Production of DNJ by Bacillus subtilis VN9 and examination alpha-glucosidase inhibitor ability of DNJ in in-vitro. Strain and Methods: Bacillus strains producing alpha-glucosidase inhibitor, optimization of culture condition for synthesizing DNJ, determination level of alpha-glucosidase inhibitor and DNJ identification. Results and Conclusions: DNJ is thermotolerant compound and strongly inhibits alpha-glucosidase, alpha-amylase, sucrase and maltase. High yield production on medium containing lactose, peptone, initial pH of 7, at 200 rpm for 4 days of fermentation.