AbstractA Plackett-Burman design was used to pre-optimize the medium composition for biobutanol production using a unique isolate of solvent-producing Clostridium YM1. Various nutrient factors affecting biobutanol production were screened using the Plackett-Burman design. These factors included: glucose, tryptone, yeast extract, peptone, ammonium acetate, KH2PO4, K2HPO4, MgSO4, FeSO4, Na2CO3, and NaCl. The results were analyzed by an analysis of variance (ANOVA), which showed that glucose, tryptone, yeast extract, peptone, K2HPO4, Na2CO3, and MgSO4 had significant effects on biobutanol production. However, ammonium acetate, KH2PO4, and FeSO4 had insignificant effects. The established model from the ANOVA analysis had a significant value of Pmodel > F = 0.0245 and an R2 value of 0.999. The estimated maximum biobutanol production was 9.01 g/L, whereas the optimized medium produced 10.93 g/L of biobutanol.