Physical and chemical differences between one-stage and two-stage hydrothermal pretreated hardwood substrates for use in cellulosic ethanol production

Background: There are many different types of pretreatment carried out to prepare cellulosic substrates for fermentation. In this study, one- and two-stage hydrothermal pretreatment were carried out to determine their effects on subsequent fermentations. The two substrates were found to behave differently during fermentation. The two substrates were then characterized using physical and chemical parameters. Results: The one-stage substrate was found to have higher carbohydrate content and lower lignin content. It exhibited a higher level of viscosity, a larger settled volume, and a slower settling time than the two-stage substrate. It also showed higher polarity and reduced crystallinity. Glycome profiling showed physical differences between the two substrates, specifically pointing toward higher levels of pectin and hemicellulose in the one-stage substrate (MS1112) as compared to the two-stage substrate (MS1107). Conclusions: We hypothesize that these physical and chemical differences between the substrates contribute to the differences seen during fermentation including: ethanol yield, ethanol titer, fermentation rate, fermentation completion time, mixing, and substrate solubilization. These findings can be used in optimizing pretreatment parameters to maximize ethanol conversion and overall process yield for hardwood substrates.