Deep eutectic solvents (DESs) are a unique category of green solvents that have gained attention in biomass processing due to their distinctive properties not offered by traditional solvents. The pH behavior of 17 selected DESs along with their temperature dependence on pH were evaluated in this study. For all investigated DESs, a temperature increase caused a decrease in pH value.

Monosaccharides such as glucose, xylose and arabinose are the main monomer units of which cellulose and hemicelluloses are composed. The cellulose and hemicelluloses content in many biomass species makes them suitable for 2G bioethanol production. Today, when 1G bioethanol production is closely monitored due to its enormous consumption of food raw materials such as wheat or corn grains, larger companies are gradually moving to pilot operations of 2G bioethanol production. However, cellulose and hemicelluloses contained in biomass are only very slightly accessible to enzymes used in 2G bioethanol production. Therefore pretreatment methods such as steam explosion are very suitable to use for fractionation of cell structure. In this paper, we tested the cellulose accessibility. We compared the cellulose accessibility of wheat straw particles with wooden particles obtained from beech and poplar. Particle size was less than 0.7 mm. We identified the optimal conditions of steam explosion pretreatment at reaction temperature of 200 °C for wheat straw, poplar and beech wood particles. The main indicator of accessibility was concentration of monomers obtained from enzymatic hydrolysis. The concentration of monomer was determined by high performance liquid chromatography. The experimental results showed different accessibility measure for each type of biomass species.

The effect of steam explosion on enzymatic hydrolysis of various parts of poplar tree (heartwood, sapwood and 1-year coppice) was investigated. These parts were milled, the obtained sawdust was chemically analysed and then steam explosion of 0.7 mm poplar particles at temperature of 205°C was performed. Concentration of monomers obtained after enzymatic hydrolysis was considered as the main indicator for cellulose accessibility. Analysis of high performance liquid chromatography showed that non-treated poplar sawdust does not enable sufficient cellulose accessibility, while excessively high temperature and rapid pressure release resulted in substantial breakdown of polysaccharides and lignin and formation of inhibitors. The concentration of monomers increased gradually in the order of coppice, sapwood and heartwood. Steam exploded heartwood gave the maximum monosaccharides concentration of 90.0 g. L-1 after 72 hours of enzymatic hydrolysis. However, glucose concentration culminated after 48 hours of this hydrolysis. This corresponds to the best holocellulose accessibility for enzymes. The maximum concentration of inhibitors (9.3 g. L-1) was determined for poplar coppice after 24 hours of enzymatic hydrolysis.

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A database for 3D structures of pepsin-like enzymes has been created on the basis of a novel approach using the Internal Coordinate System (ICS). It allows rapid comparison of multiple structures of pepsin-like enzymes without the need for preliminary calculations. Atomic displacements measured by this approach are very close to those estimated by the superposition procedures widely employed in comparing three-dimensional structures of proteins. Any new structure of pepsin-like enzyme converted to the ICS automatically becomes superimposed with all structures in a database. The ICS approach can be used for any class of enzymes and is especially efficient for families containing a large number of homologous structures.