The industrial production of sugar syrups from lignocellulosic materials requires the conduction of the enzymatic hydrolysis step at high-solids loadings (i.e., with over 15% solids [w/w] in the reaction mixture). Such conditions result in sugar syrups with increased concentrations and in improvements in both capital and operational costs, making the process more economically feasible. However, this approach still poses several technical hindrances that impact the process efficiency, known as the “high-solids effect” (i.e., the decrease in glucan conversion yields as solids load increases). The purpose of this review was to present the findings on the main limitations and advances in high-solids enzymatic hydrolysis in an updated and comprehensive manner. The causes for the rheological limitations at the onset of the high-solids operation as well as those influencing the “high-solids effect” will be discussed. The subject of water constraint, which results in a highly viscous system and impairs mixing, and by extension, mass and heat transfer, will be analyzed under the perspective of the limitations imposed to the action of the cellulolytic enzymes. The “high-solids effect” will be further discussed vis-à-vis enzymes end-product inhibition and the inhibitory effect of compounds formed during the biomass pretreatment as well as the enzymes’ unproductive adsorption to lignin. This review also presents the scientific and technological advances being introduced to lessen high-solids hydrolysis hindrances, such as the development of more efficient enzyme formulations, biomass and enzyme feeding strategies, reactor and impeller designs as well as process strategies to alleviate the end-product inhibition. We surveyed the academic literature in the form of scientific papers as well as patents to showcase the efforts on technological development and industrial implementation of the use of lignocellulosic materials as renewable feedstocks. Using a critical approach, we expect that this review will aid in the identification of areas with higher demand for scientific and technological efforts.

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Cellulose nanocrystals (CNCs), a biomaterial with high added value, were obtained from pure cellulose, Eucalyptus holocellulose, unbleached Kraft pulp, and sugarcane bagasse, by fibrillating these biomass substrates using wet disk milling (WDM) followed by enzymatic hydrolysis using endoglucanase/β-glucosidase. The hydrolysis experiments were conducted using the commercial enzyme OptimashBG or a blend of Pyrococcus horikoshii endoglucanase and Pyrococcus furiosus β-glucosidase. The fibrillated materials and CNCs were analyzed by X-ray diffraction, atomic force microscopy, scanning electron microscopy, and the specific surface area (SSA) was measured. WDM resulted in the formation of long and twisted microfibers of 1000-5000 nm in length and 4-35 nm in diameter, which were hydrolyzed into shorter and straighter CNCs of 500-1500 nm in length and 4-12 nm in diameter, with high cellulose crystallinity. Therefore, the CNC's aspect ratio was successfully adjusted by endoglucanases under mild reaction conditions, relative to the reported acidic hydrolysis method.

This study presents data on the production, purification, and properties of a thermostable β-xylanase produced by an Aspergillus awamori 2B.361 U2/1 submerged culture using wheat bran as carbon source. Fractionation of the culture filtrate by membrane ultrafiltration followed by Sephacryl S-200 and Q-Sepharose chromatography allowed for the isolation of a homogeneous xylanase (PXII-1), which was 32.87 kDa according to MS analysis. The enzyme-specific activity towards soluble oat spelt xylan, which was found to be 490 IU/mg under optimum reaction conditions (50°C and pH 5.0-5.5), was 17-fold higher than that measured in the culture supernatant. Xylan reaction products were identified as xylobiose, xylotriose, and xylotetraose. K (m) values (mg ml(-1)) for soluble oat spelt and birchwood xylan were 11.8 and 9.45, respectively. Although PXII-1 showed 85% activity retention upon incubation at 50 °C and pH 5.0 for 20 days, incubation at pH 7.0 resulted in 50% activity loss within 3 days. PXII-1 stability at pH 7.0 was improved in the presence of 20 mM cysteine, which allowed for 85% activity retention for 25 days. This study on the production in high yields of a remarkably thermostable xylanase is of significance due to the central role that this class of biocatalyst shares, along with cellulases, for the much needed enzymatic hydrolysis of biomass. Furthermore, stable xylanases are important for the manufacture of paper, animal feed, and xylooligosaccharides.