Rheological Properties of Fish and Mammalian Gelatin Hydrogels as Bases for Potential Practical Formulations.

Hydrogels have the ability to retain large amounts of water within their three-dimensional polymer matrices. These attractive materials are used in medicine and the food industry; they can serve as the basis for structured food products, additives, and various ingredients. Gelatin is one of widely used biopolymers to create hydrogels that exhibit biocompatibility and tunable rheological properties. In this study, we offer a comparative analysis of rheological properties of gelatin-based hydrogels (C = 6.67%), including mammalian gelatins from bovine and porcine skins and fish gelatins from commercial samples and samples extracted from Atlantic cod skin. Mammalian gelatins provide high strength and elasticity to hydrogels. Their melting point lies in the range from 22 to 34 °C. Fish gelatin from cod skin also provides a high strength to hydrogels. Commercial fish gelatin forms weak gels exhibiting low viscoelastic properties and strength, as well as low thermal stability with a melting point of 7 °C. Gelatins were characterized basing on the analysis of amino acid composition, molecular weight distribution, and biopolymer secondary structure in gels. Our research provides a unique rheological comparison of mammalian and fish gelatin hydrogels as a tool for the re-evaluation of fish skin gelatin produced through circular processes.

Underused Marine Resources: Sudden Properties of Cod Skin Gelatin Gel.

The main object of this work was to characterize the structure and properties of laboratory-made fish gelatin from cod skin in comparison with known commercial gelatins of fish and mammalian origin. This is one way we can contribute to the World Food Program and characterize foodstuff resources from alternative natural sources. Our research was based on the combination of an expanded set of complementary physical-chemical methods to study the similarities and distinctions of hydrogels from traditional and novel gelatin sources from underused marine resources. In this work, we have compared the morphology, supramolecular structure and colloid properties of two commercial (mammalian and fish) gelatins with gelatin we extracted from cold-water cod skin in laboratory conditions. The obtained results are novel, showing that our laboratory-produced fish gelatin is much closer to the mammalian one in terms of such parameters as thermal stability and strength of structural network under temperature alterations. Especially interesting are our experimental observations comparing both fish gelatins: it was shown that the laboratory-extracted cod gelatin is essentially more thermally stable compared to its commercial analogue, being even closer in its rheological properties to the mammalian one.

Recent Advances in Protein-Protein Interactions.

Protein-protein interactions (PPIs) lead to formation of complexes and aggregates between a pair or multiple protein molecules [...].

Highlighting the difference in nanostructure between domain-forming and domainless protic ionic liquids.

Nanoheterogeneity in some ionic liquids is a known phenomenon, but quantifying or sometimes even identifying it is not a straightforward task. We compared several known and suggested some novel approaches to identify and characterize domain segregation using the results of atomistic simulations. 10 ammonium-based protic ionic liquids with different propensity to form segregated polar and apolar domains as suggested by experimental studies were considered. They include butyl-, propyl-, 2-methoxyethylammonium nitrate, butyl- and propylammonium hydrogen sulfate, butylammonium thiocyanate (domain-forming liquids), ethylammonium and pyrrolidinium nitrate (weakly pronounced segregation), methylammonium and 2-hydroxyethylammonium nitrate (domainless liquids). Molecular dynamics simulations were performed using models based on the OPLS-AA force field with scaled ion charges. Results show that domains can be recognized and the characteristic domain length scale can be determined from peaks of Ripley's functions, peaks and large-period oscillations of finite-volume radial distribution function integral, or difference of such integrals for polar and apolar atoms, and peaks of local atom density variance. These peaks disappear with increasing temperature due to the disruption of segregated domains. In domain-forming liquids, apolar atoms are more homogeneously distributed in space than polar atoms. In addition, the probability of molecular-sized cavity formation is significantly higher in apolar domains, which determines better solubility of apolar species in domain-forming ILs. The suggested approaches can be applied to various nanostructured liquids including both ionic and molecular solvents and mixtures, as well as other systems with mesoscale ordering.

Fibril fragments from the amyloid core of lysozyme: An accelerated molecular dynamics study.

Protein aggregation and formation of amyloid fibrils are associated with many diseases and present a ubiquitous problem in protein science. Hen egg white lysozyme (HEWL) can form fibrils both from the full length protein and from its fragments. In the present study, we simulated unfolding of the amyloidogenic fragment of HEWL encompassing residues 49-101 to study the conformational aspects of amyloidogenesis. The accelerated molecular dynamics approach was used to speed up the sampling of the fragment conformers under enhanced temperature. Analysis of conformational transformation and intermediate structures was performed. During the unfolding, the novel short-living and long-living ß-structures are formed along with the unstructured random coils. Such ß-structure enriched monomers can interact with each other and propagate into fibril-like forms. The stability of oligomers assembled from these monomers was evaluated in the course of MD simulations with explicit water. The residues playing a key role in fibril stabilization were determined. The work provides new insights into the processes occurring at the early stages of amyloid fibril assembly.

Fast scanning calorimetry of lysozyme unfolding at scanning rates from 5 K/min to 500,000 K/min.

BACKGROUND: Protein denaturation is often studied using differential scanning calorimetry (DSC). However, conventional instruments are limited in the temperature scanning rate available. Fast scanning calorimetry (FSC) provides an ability to study processes at much higher rates while using extremely small sample masses [ng]. This makes it a very interesting technique for protein investigation. METHODS: A combination of conventional DSC and fast scanning calorimeters was used to study denaturation of lysozyme dissolved in glycerol. Glycerol was chosen as a solvent to prevent evaporation from the micro-sized samples of the fast scanning calorimeter. RESULTS: The lysozyme denaturation temperatures in the range of scanning rates from 5 K/min to ca. 500,000 K/min follow the Arrhenius law. The experimental results for FSC and conventional DSC fall into two distinct clusters in a Kissinger plot, which are well approximated by two parallel straight lines. CONCLUSIONS: The transition temperatures for the unfolding process measured on fast scanning calorimetry sensor are significantly lower than what could be expected from the results of conventional DSC using extrapolation to high scanning rates. Evidence for the influence of the relative surface area on the unfolding temperature was found. GENERAL SIGNIFICANCE: For the first time, fast scanning calorimetry was employed to study protein denaturation with a range of temperature scanning rates of 5 orders of magnitude. Decreased thermal stability of the micro-sized samples on the fast scanning calorimeter raise caution over using bulk solution thermal stability data of proteins for applications where micro-sized dispersed protein solutions are used, e.g., spray drying.

Molecular dynamics study of unfolding of lysozyme in water and its mixtures with dimethyl sulfoxide.

All-atom explicit solvent molecular dynamics was used to study the process of unfolding of hen egg white lysozyme in water and mixtures of water with dimethyl sulfoxide at different compositions. We have determined the kinetic parameters of unfolding at a constant temperature 450K. For each run, the time of disruption of the tertiary structure of lysozyme tu was defined as the moment when a certain structural criterion computed from the trajectory reaches its critical value. A good agreement is observed between the results obtained using several different criteria. The secondary structure according to DSSP calculations is found to be partially unfolded to the moment of disruption of tertiary structure, but some of its elements keep for a long time after that. The values of tu averaged over ten 30ns-long trajectories for each solvent composition are shown to decrease very rapidly with addition of dimethyl sulfoxide, and rather small amounts of dimethyl sulfoxide are found to change the pathway of unfolding. In pure water, despite the loss of tertiary contacts and disruption of secondary structure elements, the protein preserves its compact globular state at least over 130ns of simulation, while even at 5mol percents of dimethyl sulfoxide it loses its compactness within 30ns. The proposed methodology is a generally applicable tool to quantify the rate of protein unfolding in simulation studies.

New insights into the solubility of graphene oxide in water and alcohols.

One of the main advantages of graphene oxide (GO) over its non-oxidized counterpart is its ability to form stable solutions in water and some organic solvents. At the same time, the nature of GO solutions is not completely understood; the existing data are scarce and controversial. Here, we demonstrate that the solubility of GO, and the stability of the as-formed solutions depend not just on the solute and solvent cohesion parameters, as commonly believed, but mostly on the chemical interactions at the GO/solvent interface. By the DFT and QTAIM calculations, we demonstrate that the solubility of GO is afforded by strong hydrogen bonding established between GO functional groups and solvent molecules. The main functional groups taking part in hydrogen bonding are tertiary alcohols; epoxides play only a minor role. The magnitude of the bond energy values is significantly higher than that for typical hydrogen bonding. The hydrogen bond energy between GO functional groups and solvent molecules decreases in the sequence: water > methanol > ethanol. We support our theoretical results by several experimental observations including solution calorimetry. The enthalpy of GO dissolution in water, methanol and ethanol is -0.1815 ± 0.0010, -0.1550 ± 0.0012 and -0.1040 ± 0.0010 kJ g-1, respectively, in full accordance with the calculated trend. Our findings provide an explanation for the well-known, but poorly understood solvent exchange phenomenon.

Quantitative description of the hydrophobic effect: the enthalpic contribution.

A new method of experimental determination of the hydrophobic effect enthalpy is proposed. The method is based on regarding the hydration enthalpy as the sum of the nonspecific hydration enthalpy, specific hydration enthalpy, and the hydrophobic effect enthalpy. The hydrophobic effect enthalpies of noble and simple substance gases, alkanes, arenes, and normal aliphatic alcohols are determined. For the noble gases and alkanes, the hydrophobic effect enthalpy is found to be negative and independent of the size of molecule. For aromatic hydrocarbons, it is positive and grows up with the size of the hydrocarbon. The hydrophobic effect enthalpies of normal aliphatic alcohols are determined by assuming that the specific interaction enthalpies of alcohols in water and in methanol are equal. The hydrophobic effect enthalpy values for the aliphatic alcohols (-10.0 +/- 0.9 kJ.mol(-1)) were found to be close to the alkanes hydrophobic effect enthalpies (-10.7 +/- 1.5 kJ.mol(-1)).