Roberto Valenzuela

Steam explosion pretreatment is suitable for bioethanol production from Eucalyptus globulus wood. Multifactorial experiment designs were used to find the optimal temperature and residence time required to obtain the best glucose yield from the enzymatic hydrolysis of pretreated materials. The chemical composition, crystallinity index, morphology and polymerization degree of the pretreated materials were correlated with enzymatic accessibility. Simultaneous saccharification and fermentation (SSF) using a fed-batch strategy was applied to three different laboratory-scale fermenters. The optimization of the pretreatment was obtained at 208 °C and 11 min. However, the enzymatic hydrolysis performance did not show significant differences from the material obtained at 196 °C and 9.5 min, which was determined to be the real optimum, owing to its lower energy requirement. The vertical fermenter with type “G” blades and the horizontal fermenter with helical blades were both highly efficient for reaching ethanol yields close to 90% based on dry wood, and ethanol concentrations close to 9.0% v/v.

In order to understand the relation between chemical composition, microscopic structure and enzymatic digestibility, different Eucalyptus globulus wood pretreated samples were examined. Pretreated materials obtained by steam explosion and autohydrolysis were compared with those obtained by organosolv and kraft processes. Chemical analyses of pretreated materials showed a decrease in the content of xylans, except in the kraft pulp. FT-IR spectra showed that the residual lignin in autohydrolysis pulp had experienced greater changes compared to those in steam explosion and organosolv pulps, whereas minor changes in lignin kraft pulp were observed. The fiber morphology indicated that autohydrolysis pretreatment was the most aggressive treatment. Reduction in the content of lignin and its redistribution on the fiber wall were confirmed through confocal laser microscopy. The formation of discrete lignin droplets deposited on the surface of the fibers was observed in all pretreatments, with a higher frequency in organosolv followed by steam explosion. A significant increase in enzymatic accessibility was achieved in organosolv, autohydrolysis and steam explosion pulps, due to xylans removal combined with lignin redistribution. Homogeneous lignin distribution and higher xylan content may be related to the low enzymatic hydrolysis efficiency in kraft pulp.

β-Glucosidase from Trichoderma reesei was immobilised on synthetic superparamagnetic magnetite (Fe₃O₄) nanoparticles with a mean diameter of 10 nm and were used to supplement cellulase in the enzymatic hydrolysis of three substrates: wheat straw pretreated by steam explosion, Eucalyptus globulus pretreated by hydrothermolysis and E. globulus pretreated by hydrothermolysis followed by alkaline extraction. The hydrolysis yields for each pretreated material, using immobilised β-glucosidase (I-βG) and free cellulase, were 76.1%, 83.6% and 75.6%, respectively, and resulted in an improved hydrolysis yields compared with only cellulase. These yields were at most 10% lower than yields reached with free enzymes. The (I-βG) was magnetically recovered and successfully reused twice. The differences in the hydrolysis yields were not significant (p > 0.05) in the case of steam-exploded wheat straw and E. globulus pretreated by hydrothermolysis followed by alkaline extraction. The immobilisation of enzymes provides an opportunity to reduce the costs of enzymes in the bioethanol production process.