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Soybean meal is a by-product from soybean oil manufacturing. It contained, on the mass basis, crude fiber (6.3%) and protein (47.6%) after oil extraction. The aim of study was to investigate the cultivation of isolated Clostridium sp. G10 using soybean meal hydrolysate as a N- and C-source in fermentation medium. Soybean meal hydrolyzed with 0.02 to 0.1 M of HCl and H2SO4 and was heated at 121 °C, 15 psi for 10-30 min. The result showed that hydrolyzing soybean with 0.1 M HCl and 20-min heating period were the optimal conditions to obtain soluble protein (24.45 ±2.21 g/L) and reducing sugar (13.12 ±1.09 g/L). To study the effect of N- and C-sources replacement in T6 medium, soybean meal hydrolysate was utilized to replace yeast extract and tryptone resulting in 8 g/L soluble protein and used as a C-source with the addition of glucose with 50 g/L reducing sugar equivalent. The butanol concentration (10.80 ±0.44 g/L) was slightly lowered than that of control T6 medium (12.15 ±0.07 g/L). These results demonstrated that Clostridium sp. was able to uptake the N-and C-sources contained in the hydrolyzed soybean meal. However, a C-source alternative is required to mitigate the cost of raw materials., Proceedings of the 30th European Biomass Conference and Exhibition, 9-12 May 2022, Online, pp. 927-931