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Microbial inhibitors including weak acids, furan derivatives and phenolic compounds are key problems of cellulosic bio-fuels production by fermentation. Most of these inhibitors are sugars and lignin degradation compounds, which are almost unavoidable during pretreatment processes. While, most of the one stage pretreatment has been conducted at high severity factors of 3.5 or more to get high sugar yield, with increase in severity factor, high concentration of microbial inhibitors were formed and significantly affected downstream biofuel yield. Thus, a two-stage pretreatment of corn stalk, hydrothermal followed by oxalic acid, under low severity factor and its enzymatic degradability was investigated in this study to identify fermentable sugar production and corresponding microbial inhibitors formation. Additionally, effect of equivalent severity factors of 2 to 3.5 and particle sizes of 1 to 35 mm were also studied systematically. Particle size of 15 mm was found as an optimum size at an equivalent severity factor of 2.5. Sugars 61.99 ± 0.03 g and inhibitors 5.12 ± 0.01 g from 100 g of corn stalk were obtained at the optimum particle size and pretreatment condition. The highest glucan conversion and recovery at the optimum conditions were 92.95± 0.08 and 78.42± 0.07%, respectively. Overall, the two-stage pretreatment process with the larger particle size and low equivalent severity factor could be an alternative to reduce microbial inhibitors formation and excessive biomass processing cost. Key words: Bio-fuel, corn stalk, pretreatment, particle size, microbial inhibitors, fermentable sugars.

Fermentable sugars and microbial inhibitors formation from two-stage pretreatment of corn stalk with variation in particle size and severity factor