Creation of green chemical substances and biofuels in biorefineries may be the potential choice for petrochemicals and fuel in transitioning of petro-economy into bioeconomy. biomass and their issues, besides this strategic function of nano and Formononetin (Formononetol) biotechnological strategies to the sustainability and viability of biorefineries can Formononetin (Formononetol) be discussed. respectively, attributing this known reality to removing lignin, incomplete hemicellulose solubilization, and cellulose retention. As amorphous fractions from the materials are CDK6 simpler to remove during pretreatment, the full total crystallinity from the materials shall boost, however the crystallinity of staying cellulose could be lower if in comparison to neglected biomass, and, hence, the pretreatment leading to higher enzymatic digestibility (Driemeier et al. 2011). The Formononetin (Formononetol) scholarly research of biomass features and Formononetin (Formononetol) its own adjustments during pretreatment, aswell as the knowledge of the connections of factors (e.g.: lignin removal and raising of surface) are key for the introduction of brand-new technicals and circumstances aswell as process marketing of known methodologies. Usually, the crystallinity index (CI) is one of the most applied methods to verify changes in the biomass crystallinity related to the pretreatments. However, some authors reported that this method could not be effective due to the difficulties to distinguish the specific crystallinity of the cellulose and total biomass. This fact was discussed by Driemeier et al. (2011), which observed the evolution of cellulose crystals from sugarcane bagasse after pretreatment by hydrothermal, dilute acid or steam explosion methods, and soda delignification. Those authors observed a decrease in crystal-to-cellulose ratio after pretreatment, an effect opposite to preferential removal of non-crystalline cellulose. The observed behavior was explained by a cellulose partial decrystallization or more defective crystallites as a result of the treatments. As an alternative to evaluate the effect of pretreatment, Bernardinelli et al. (2015) demonstrated the application of cross polarization by multiple contact periods (Multi-CP) to obtain quantitative 13?C solid-state nuclear magnetic resonance (SSNMR) spectra to evaluate raw and pretreated sugarcane bagasse. This method was reported as more feasible to unravel different pretreatments action in biomass cell wall digestion changing cellulose ultrastructure. Actually, aiming to increase scientific comprehension of biomass recalcitrance, researchers have studied changes in the structural morphology Formononetin (Formononetol) of biomass along with lignocellulosic pretreatments by different strategies. Chandel et al. (2014) evaluated sequential acidCbase pretreatment, aiming to first obtain hemicellulosic hydrolysate, followed by lignin solubilization of remaining solid portion by alkali treatment. Thus, cellulose in the remaining portion was cleaved in hexose monomers sugars by enzymatic hydrolysis. A large number of structural changes were observed in biomass along pretreatments using different physical analysis. For instance, after acid hydrolysis, 92.78% hemicellulose was removed, increasing the cellulose amount and, hence, the crystallinity of the sample. By light microscopic analysis, thin shape of particles and more cylindrical shape (20?m size), as well as the relocation of lignin portion on the surface, were found compared to native sugarcane bagasse. In addition, the relation of cellulose/hemicellulose bands was verified by Raman spectroscopy. Even by studying the modifications in composition and structure of biomass due to pretreatment, there is not a perfect and unique indicator of biomass recalcitrance or even to be used like a predictor of pretreatment achievement. Nevertheless, there are a few tries with this true way. Costa et al. (2013) reported a report about enzymatic hydrolysis of internodes of sugarcane hybrids with differing lignin material. Those authors noticed a correlation between your chemical composition as well as the microscopy features of the cross sugarcane internode fractions using the efficiency from the enzymatic hydrolysis. A quadratic polynomial equation was adjusted when enzymatic.