Anna Bannikova

Structural behaviour in condensed bovine serum albumin systems following application of high pressure.

The present study shows that application of high hydrostatic pressure of 600MPa for 15min at ambient temperature on condensed bovine serum albumin systems (BSA) with up to 80% w/w solids content has a limited effect on the conformational structure of the protein, as compared to thermal treatment. This was demonstrated throughout the experimental concentration range using small-deformation dynamic oscillation, differential scanning calorimetry and infrared spectroscopy. BSA possesses seventeen disulfide linkages per molecule, which constitutes a stable arrangement with high energy requirements for substantial structure alteration. Upon cooling, pressurised materials undergo vitrification and networks exhibit comparative mechanical strength to that of thermally treated counterparts. The mechanical manifestation of the glass transition region and glassy state for atmospheric and pressurised samples was examined by the method of reduced variables and the combined framework of WLF/free volume theory producing disparate predictions of the glass transition temperature for the two types of polymeric network.

Release mechanism of omega-3 fatty acid in κ-carrageenan/polydextrose undergoing glass transition

A high-solid matrix of κ-carrageenan with polydextrose was developed to entrap α-linolenic acid, which is an omega-3 bioactive compound. Physicochemical analysis of this system utilised modulated DSC, dynamic oscillation in shear, ESEM, FTIR and WAX diffraction. The carbohydrate matrix was conditioned through an extensive temperature range to induce changes in molecular morphology and identify the network glass transition temperature. Thermally induced variation in phase morphology was employed to rationalise transportation patterns of the bioactive compound within the high-solid preparation. Thus, experimental observations using UV-vis spectroscopy modelled diffusion kinetics to document the mobility arresting effect of the vitrifying matrix on the micro-constituent. Within the glass transition region, results argue that free volume theory is the molecular process governing structural relaxation. Further, Less Fickian diffusion follows well the rate of molecular transport of α-linolenic acid as a function of time and temperature of observation in the condensed matrix.

Controlled release of thiamin in a glassy κ-carrageenan/glucose syrup matrix.

The work dealt with the diffusional mobility of thiamin embedded in a high-solid matrix of κ-carrageenan with glucose syrup. It utilized thermomechanical analysis in the form of modulated differential scanning calorimetry and small-deformation dynamic oscillation in shear, Fourier transform infrared spectroscopy, wide angle X-ray diffraction, scanning electron microscopy and UV-vis spectrophotometry. The structural properties of the matrix were assessed in a temperature induced rubber-to-glass transformation. A thiamin-dye binding assay was employed to monitor the diffusion process of the vitamin from the high-solid preparation to ethylene glycol. The relationship between mechanical properties of the carbohydrate matrix and vitamin mobility was assessed via the application of the combined framework of the free volume theory and the predictions of the reaction rate theory. Results argue that the transport of the micronutrient is governed by the structural relaxation of the high-solid matrix. These were further treated with the concept of Fickian diffusion coefficient to provide the rate of the bioactive compound mobility within the present experimental settings.

Phase behaviour of gelatin/polydextrose mixtures at high levels of solids

This investigation focuses on understanding the phase behaviour of gelatin when mixed with polydextrose (co-solute) primarily at high solid concentrations. The experimental work was carried out using small deformation dynamic oscillation in shear, modulated differential scanning calorimetry, Fourier transform infrared spectroscopy, wide angle X-ray diffraction and environmental scanning electron microscopy. A progression in the mechanical strength and thermal stability of the gelatin network was observed with the addition of polydextrose to the system. Combined thermomechanical and microscopy evidence argues for the development of phase separation phenomenon between protein and co-solute in high-solid preparations, where gelatin maintains helical conformation to provide network integrity as well as glassy consistency at subzero temperature. At the high solids regime, glassy consistency was treated with theoretical frameworks from the synthetic polymer research to pinpoint the glass transition temperature of the system.

Investigation on the phase behaviour of gelatin/agarose mixture in an environment of reduced solvent quality

Investigation on the phase behaviour of a biopolymer mixture has been performed using 7.5% (w/w) gelatin and 1.5% (w/w) agarose in the presence of variable amounts of polydextrose as the co-solute from low to high levels of total solids. Mechanical observation of the system was performed using small deformation dynamic oscillation in shear along with thermal studies using modulated differential scanning calorimetry. Micrographs provided images of the changing morphology of the network with the addition of co-solute. Agarose and gelatin form non-interactive bicontinuous phases in the aqueous environment. Systematic increase in the concentration of polydextrose prevents the formation of a stable agarose network, with the polysaccharide chains dispersing in the high solids environment. Gelatin, on the other hand, retains its conformational stability even at a saturating co-solute environment through enhanced protein structuring. Vitrification studies on the high solids system at subzero temperatures provides information on the structural and molecular relaxation identified as a glass transition phenomenon. Fourier transform infrared spectroscopy was used to analyse potential direct interaction between polymers and co-solute. The extent of amorphicity in the system was confirmed using wide angle X-ray diffraction.

Networks of polysaccharides with hydrophilic and hydrophobic characteristics in the presence of co-solute.

The present investigation deals with the changing network morphology of agarose and high methoxy pectin when mixed with polydextrose as co-solute at concentrations varying up to high level of solids. Thermomechanical analysis and micro-imaging were performed using small deformation dynamic oscillation in shear, modulated differential scanning calorimetry and environment scanning electron microscopy. Fourier transform infrared spectroscopy and wide angle X-ray diffraction were practised to examine the nature of interactions between polymer and co-solute, and the extent of amorphicity of preparations. We observed a decline in the mechanical strength of aqueous agarose preparations upon addition of high levels of polydextrose, which should be attributed to reduced enthalpic content of the coil-to-helix transition of the polysaccharide network. Glass transition phenomena were observed at subzero temperatures in condensed preparations, hence further arguing for the formation of a lightly cross-linked agarose network with changing solvent quality. High levels of co-solute induce formation of weak pectin gels at elevated temperatures (even at 95°C), which with lowering temperature exhibit increasing strength. This results in the formation of rubbery pectin gels at ambient temperature, which upon controlled cooling to subzero temperatures convert to a clear glass earlier than the agarose counterparts.

Analysis on the effectiveness of co-solute on the network integrity of high methoxy pectin

Co-solute requirements for high methoxy pectin gelation were observed by the addition of glucose syrup and polydextrose at concentrations varying from 50% to 78% (w/w). Pectin content was fixed at 2% (w/w) in formulations. Studies from small deformation dynamic oscillation in shear, modulated differential scanning calorimetry and environmental scanning electron microscopy are reported. Structural properties of pectin preparations were recorded in relation to the molecular weight and concentration of added co-solute in an acidic environment (pH ∼3.0). High levels of co-solute induce formation of weak pectin gels at elevated temperatures (even at 95°C), which upon subsequent cooling exhibit increasing strength and convert to a clear glass at subzero temperatures. Fourier Transform Infrared Spectroscopy and wide angle X-ray diffraction were practised to examine the nature of interactions between polymer and co-solute and the extent of amorphicity of preparations. Glucose syrup is an efficient plasticiser leading to a reduction in the glass transition temperature (T(g)) of the pectin network, whereas polydextrose assists in the formation of stronger pectin gels in the rubbery state.

Protein‐loaded sodium alginate and carboxymethyl cellulose beads for controlled release under simulated gastrointestinal conditions

Network characteristics of sodium alginate and carboxymethyl cellulose, in relation to the controlled release of bovine serum albumin under simulated gastrointestinal conditions, have been examined. Small-deformation dynamic oscillation in-shear, compression testing and optical microscopy were employed to monitor the structural characteristics of polysaccharide beads as excipients for protein entrapment. Work focused on gastrointestinal functionality and controlled release utilising variable acidity and counterion addition in preparations. Beads were found to shrink in gastric and swell in intestinal conditions, thus emphasising the disparate effect of experimental conditions. Changes in mechanical properties reflected alterations in gel microstructure, with beads becoming relatively strong in the gastric environment. In contrast, networks were weakened in the intestinal environment resulting in bead disintegration towards the end of digestion time. This type of structural behaviour allows the controlled delivery of the protein, as a model bioactive, in a particular part of the gastrointestinal tract.

Structural and Molecular Properties of Condensed Globular Proteins from High Pressure Perspectives

The marked increase in the application of high pressure processing in the food industry worldwide created a range of novel products with enhanced natural flavour and retention of natural vitamins compared to thermally treated products. The present treatise deals with the effect of high hydrostatic pressure on the structural properties of globular proteins with potential industrial interest in a wide range of high-solid foods. Globular proteins exhibit glassy behaviour at subzero temperatures in high-solid preparations (e.g. 80%, w/w) recorded rheologically and modelled theoretically. DSC and FTIR results revealed that disulphide bonds are involved in the pressure stability of globular proteins. BSA comprises 17 disulphide linkages being the most stable at high pressure, whereas whey protein with 2 disulphide bonds is the most affected globular protein under high pressure (600 MPa for 15 min at ambient temperature). However, ovalbumin with one disulphide linkage does not follow this sequence, an outcome that raises the issue of the effect of surface hydrophobicity of the molecule. Thus, ovalbumin has a hydrophobicity value (So) of around 100, which falls between that of BSA (So ∼ 2200) and whey protein (So ∼ 35), indicating that both phenomena, i.e. disulphide linkages and surface hydrophobicity, are combined in the observed structural properties of globular proteins following application of high pressure. This may have industrial significance in relation to the formulation and stabilisation of “functional food” products as well as in protein ingredients and concentrates by replacing spray dried powders with condensed HPP-treated pastes that maintain techno- and biofunctionality.