The researchers optimized Organosolv pretreatment conditions to remove lignin and release polysaccharides from rice straw for biofuel production. By testing different temperature, time, and acid concentration levels, they found the best conditions to be 75.41°C, 29.68 minutes, and 69.85% acid concentration. This resulted in 73.17% lignin removal, 46.62% hemicellulose solubilization, and 74.09% cellulose release. The study showed that all process parameters significantly affected the breakdown of rice straw components, with time having the strongest impact. The combination of Organosolv pretreatment and Bacillus sp. BMP01 hydrolysis was effective in increasing sugar and ethanol yields from rice straw.