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In this study, we intended to increase mass production of bacillus subtilis through medium optimization by statistical experimental method. For the seed culture of Bacillus subtilis, the basic medium and inoculum age was tryptic soy broth and 15 hours, respectively. Final dry cell weight and protease contents of Bacillus subtilis by basic production medium were 0.27 g/L, 0.04 U/mL, respectively. First, nutrient sources were selected starch, complex nitrogen(yeast extract & casitone) and ammonium phosphate for mass production of Bacillus subtilis by using one factor at a time. Dry cell weight and protease contents by obtained production medium were increased 20 and 19 times, respectively, as compared to those determined with basic production medium. Plackett-Burman experimental design was used to determine the key factors which are
critical to produce the biological products by the fermentation. According to the result of Plackett-Burman experimental design, key factors were selected starch, complex nitrogen(yeast extract & casitone) and Na2CO3. To obtained the optimum concentrations of three variables we are performing response surface methodology.