The role of water in the reversibility of thermal denaturation of lysozyme in solid and liquid states.

Although unfolding of protein in the liquid state is relatively well studied, its mechanisms in the solid state, are much less understood. We evaluated the reversibility of thermal unfolding of lysozyme with respect to the water content using a combination of thermodynamic and structural techniques such as differential scanning calorimetry, synchrotron small and wide-angle X-ray scattering (SWAXS) and Raman spectroscopy. Analysis of the endothermic thermal transition obtained by DSC scans showed three distinct unfolding behaviors at different water contents. Using SWAXS and Raman spectroscopy, we investigated reversibility of the unfolding for each hydration regime for various structural levels including overall molecular shape, secondary structure, hydrophobic and hydrogen bonding interactions. In the substantially dehydrated state below 37 wt% of water the unfolding is an irreversible process and can be described by a kinetic approach; above 60 wt% the process is reversible, and the thermodynamic equilibrium approach is applied. In the intermediate range of water contents between 37 wt% and 60 wt%, the system is phase separated and the thermal denaturation involves two processes: melting of protein crystals and unfolding of protein molecules. A phase diagram of thermal unfolding/denaturation in lysozyme - water system was constructed based on the experimental data.

Polyamide-amine-epichlorohydrin (PAE) induced TiO2 nanoparticles assembly in cellulose network.

HYPOTHESIS: TiO2-NPs-Cellulose composites functionality depends on the retention and dispersion of NPs in the composites. SAXS and SEM can be combined to reveal the effect PAE has on the NPs aggregation, retention and interaction mechanisms in the composites. EXPERIMENTS: TiO2-NPs-Cellulose sheets were made by first preparing PAE-cellulose suspensions of different PAE dosages (10 and 50 mg of PAE/g fibres). The TiO2 NPs suspension (at different NPs loading) was then added to the cellulose-PAE suspension. The final suspension was used to make flexible paper-like composites sheets. SEM and SAXS quantified NPs retention and aggregation state. FINDINGS: PAE dosage of 20 mg/g cellulose provides full surface coverage of cellulose fibres. A 10 mg of PAE/g cellulose covers half the cellulose surface area and no free PAE remains in the suspension. PAE dosage of 50 mg/g cellulose gives full cellulose surface coverage and provides a large amount of PAE (30 mg/g cellulose) free in the suspension. Surprisingly, at both PAE dosages, NP coagulates and the size of the aggregates increase with NPs loading. Aggregates of two particle sizes (10 and 35 nm) are formed and the number density of smaller particles is higher than larger particle. The NPs aggregates and their retention are similar at both PAE dosages, which is explained by different PAE-NPs bridging mechanisms.