Eucalyptus globulus wood was subjected to autohydrolysis pretreatment at different severity factors. The pretreated materials were enzymatically saccharified at a substrate load of 10% (w/v) using a cellulase enzyme complex. Around 82-95% of original glucans were retained in the pretreated material, and the enzymatic hydrolysis yields ranged from 58% to 90%. The chemical and structural changes in the pretreated materials were investigated by microscopic (SEM, LSCM) and spectroscopic (2D-HSQC NMR and FT-IR) techniques. 2D-NMR results showed a reduction in the amounts of β-O-4 aryl-ether linkages and suggested the presence of newly condensed structures of lignin in the biomass pretreated at the more severe conditions. Furthermore, the microscopic analysis showed that lignin migrates out of the cell wall and re-deposits in certain regions of the fibers at the more severe conditions to form droplet-like structures and expose the cellulose surface. These changes improved the glucose yield up to 69%, on dry wood basis.
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.
Eucalyptus globulus wood chips were subjected to autohydrolysis pretreatment at 175ºC at three different residence times. Part of the recovered solids were submitted to alkaline extraction with NaOH solution to remove leachable lignin. The chemical composition of the fibrous material was analyzed by HPLC, Py-GCMS and 2D-NMR HSQC, while morphological changes were evaluated by SEM and LSCM. The pretreated materials were hydrolyzed with cellulases at a substrate loading of 10% (w/v) for up to 72 h. Glucose yields (based on dry wood) obtained in the enzymatic hydrolysis ranged between 38% and 65%, depending on reaction time in the autohydrolysis pretreatment. After the alkaline extraction, no significant change was observed in the yields in the enzymatic hydrolysis at 72 h, but at the lower severities, the initial rates of saccharification increased. The main effect of the hydrothermal pretreatment was removal of hemicelluloses, resulting in enriched cellulose pulps. SEM and LSCM images of the hydrothermal pretreated samples showed a disruption of the fiber surface, mainly in those samples obtained at the higher severity. Py-GC/MS and HSQC analysis showed that no major changes in the lignin structure occurred in the samples obtained by autohydrolysis and further alkaline extraction. By autohydrolysis at the higher severity (So=4.02), the lateral chains in lignin were cleaved and the formation of lignin droplets was observed. Hemicelluloses removal and lignin redeposition as droplets in certain regions of the fiber surface was associated with the higher accessibility of cellulose and the yield increase of the enzymatic hydrolysis.
The objective of this study was to investigate structural changes and lignin redistribution in Eucalyptus globulus pre-treated by steam explosion under different degrees of severity (S0), in order to evaluate their effect on cellulose accessibility by enzymatic hydrolysis. Approximately 87.7% to 98.5% of original glucans were retained in the pre-treated material. Glucose yields after the enzymatic hydrolysis of pre-treated material improved from 19.4% to 85.1% when S0 was increased from 8.53 to 10.42. One of the main reasons for the increase in glucose yield was the redistribution of lignin as micro-particles were deposited on the surface and interior of the fibre cell wall. This information was confirmed by laser scanning confocal fluorescence and FT-IR imaging; these microscopic techniques show changes in the physical and chemical characteristics of pre-treated fibres. In addition, the results allowed the construction of an explanatory model for microscale understanding of the enzymatic accessibility mechanism in the pre-treated lignocellulose.
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