Avaliação do pré-tratamento de explosão a vapor catalisado por ácido cítrico e hidróxido de sódio sobre a hidrólise enzimática do bagaço de cana-de-açúcar

Abstract

Nowadays, there is various types of studied pre-treatments to reduce the lignocellulosic biomass recalcitrance in order to increase its chemical / enzymatic digestibility, so that it can be used in ethanol production and / or other bioproducts value. This study evaluated the effect of steam explosion pretreatment catalyzed by citric acid and sodium hydroxide, and alkaline delignification pretreatment over chemical and structural properties of sugarcane bagasse, as well as the enzymatic hydrolysis process. Steam explosion pretreatment was conducted in a 1.4 L reactor at temperature of 180 °C and 5 min hold time. The delignification step with NaOH (2% m:m) was performed at 120 °C under reflux for 4 hours. Chemical and structural characterization of raw and pretreated biomass was performed by FTIR, SEM and XRD. Enzymatic hydrolysis was performed with final volume of 20 mL consisting 3% SCB on dryweight basis, sodium citrate buffer 50 mM (pH = 5.0) and 10 FPU of Cellic® Ctec 3 enzyme complex. Flasks were kept under agitation of 150 rpm at 50 °C during 72 h. It was analyzed 1.5 mL aliquots after 0, 12, 24, 36, 48 and 72 hours to determinate total reducing sugars (TRS) by acid 3,5-dinitrosalicílic method (DNS). Sugarcane bagasse showed 24.22% lignin, 27.61% hemicelluloses and 42.77% of cellulose, however after steam explosion pretreatment catalyzed by citric acid, it was obtained a biomass with lesser hemicelluloses amount (16.16%) and cracks formation on fiber cell wall were observed. The bagasse pretreated by steam explosion by NaOH showed complete destructuring of fiber and lignin removal was 65%, while hemicellulosic fraction was preserved. After submitting untreated and pretreated lignocellulosic biomass by steam explosion to alkaline delignification process, complete destructuring and solubilization of the lignocellulosic matrix was observed with 85-90% lignin removal in all samples. The crystallinity index of biomass after pretreatments increased when compared to raw material and this could be associated to amorphous components removal, such as lignin and hemicelluloses, and also the amorphous cellulose fraction. The theoretical value corresponding to 100% of saccharification corresponds to approximately 33.0 g·L-1, compared to the mass percentage of biomass used in the enzymatic hydrolysis process. After 72 h enzymatic hydrolysis of raw and pretreated biomass by steam explosion, the highest concentration of total reducing sugars (TRS) and the highest percentage of saccharification were obtained for sugarcane bagasse (23.05 g·L-1, 69,15% saccharification). In in nature biomass hydrolysis, after alkaline delignification process, ART concentration increased 18.33 g·L-1, while pretreated biomass by steam explosion with water, citric acid and NaOH followed by alkaline delignification increase TRS to 19.67 g·L-1, 19.93 g·L-1 and 6.87 g·L-1. After delignification, it was noted that saccharification percentage for untreated biomass increased from 11.88% to 69.15%, while for bagasse after steam explosion and delignification this percentage was between 82.05% - 89.79%. Lastly, it should be noted that in biomass previously pretreated by NaOH steam explosion, lignin solubilization after the second pretreatment had an increase of only 20% and TRS concentration of 6.87 g·L-1. Thus, it is believed that increasing NaOH solution concentration to perform steam explosion could not need to implement delignification step.