AN ABSTRACT OF THE THESIS OF Thara G. Siddaramu, for the Master of Science degree in Civil and Environmental Engineering, presented on February 5, 2011, at Southern Illinois University Carbondale. TITLE: EVALUATION OF DIFFERENT PRETREATMENT APPROACHES FOR DISRUPTING LIGNOCELLULOSIC STRUCTURES MAJOR PROFESSOR: Dr. Yanna Liang There are two major steps in biofuel production- pretreatment of lignocellulosic materials and enzymatic hydrolysis. The present study investigated the ability of two pretreatment methods, namely traditional oven and microwave oven treatments for disrupting lignocellulosic structures. The substrates tested were Jatropha seed cake and sweet sorghum bagasse. In recent years, Jatropha curcas also known as physic nut or purging nut has attracted extensive attention due to its several unique characteristics. Similarly, sweet sorghum has the potential to provide great value to energy sectors and food industries being that the entire plant is rich in various sugars and nutrients. Both crops can adapt to various climates, and can withstand extended drought conditions compared to other crops. Additionally, both Jatropha seed cakes and sweet sorghum bagasse are good sources of lignin and carbohydrates, which could be used for production of biofuels only if the sugars can be unlocked. Several treatment methods such as mechanical, physical, chemical and biological treatments have been reported to breakdown the cellulosic structure of biomass. However, other low cost and quicker methods, such as ovenpretreatment and microwave irradiation have not been evaluated for Jatropha seed cake and Sweet Sorghum Bagasse (SSB), respectively. Composition change of Jatropha seed cake samples was evaluated upon lime pretreatment at 100 oC with different parameters. With a lime dose of 0.2 g and a water content of 10 ml per gram of cake and a treatment period of 1 h, 38.2 ± 0.6% of lignin was removed. However, 65 ± 16% of hemicellulose was also lost under this condition. For all the treatments tested, cellulose content was not affected by lime supplementation. Through further examining total reducing sugar (TRS) release by enzymatic hydrolysis after lime pretreatment, results indicated that 0.1 g of lime and 9 ml of water per gram of cake and 3 h pretreatment produced the maximal 68.9% conversion of cellulose. Without lime pretreatment, the highest cellulose conversion was 33.3%. Finally, this study shows that Jatropha seed cake samples could be hydrolyzed by enzymes. Even though the cellulose content was not high for this Jatropha cake sample, the fractionation by lime presented in this study opened the door for other applications, such as removal of lignin and toxicity for use as animal feed and fertilizer. The microwave radiation pretreatment of SSB was evaluated with or without lime (0.1 g/g bagasse) at 10 ml water/g bagasse for 4 min. TRS release over 72-h enzymatic hydrolysis was different for samples treated differently and at different solid loadings. The TRS concentration was increased by 2 and 5-fold from 0 to 24 hours in non lime-pretreated and lime-pretreated samples, respectively. Further incubation of samples for 48 and 72 h did not result in increased TRS. Comparing different solid loadings of samples treated with or without lime, 1% solid content resulted in 1.4 times higher TRS increase than that of 5% solid concentration. Therefore, lime was effective in disintegrating lignocellulosic structures and making cellulose more accessible for saccharification. Higher solid loadings which can lead to higher sugar concentrations are desired for downstream biofuel production. But, as shown in this study, higher concentration of bagasse samples decreased rate of cellulose hydrolysis due to poorer mixing efficiency and hindrance to interactions between enzymes and solid materials. Thus, an optimal solid content needs to be determined for maximal cellulose hydrolysis and for preparing the hydrolysates for downstream processes, either bioethanol or lipid production.

Hydrolysis of cellulosic biomass is an important and ongoing subject of research due to generating precursors for biofuel synthesis. This work involves hydrolysis of microcrystalline cellulose and Arundo donax to glucose. Methods for acid hydrolysis include the use of sulfuric acid, phosphomolybdic acid, blue molybdenum using low concentrations at 40-100 °C. For comparison purposes, enzymatic hydrolysis was also carried out using cellulase. Products were quantified by measuring total organic carbon and reducible sugars using dinitrosalicylic acid. Use of phosphomolybdic acid, blue molybdenum and sulfuric acid even at 6.0 M did not increase the amount of glucose compared to using water only. Interestingly, enzymatic hydrolysis of powdered Arundo donax without pretreatment was successful and resulted in statistically similar amounts of glucose compared to using microcrystalline cellulose. Efforts are ongoing to understand the enzyme kinetics in the hydrolysis of Arundo donax and potentially increase the yield of glucose using chemical and microbiological pretreatment.