Microscopic investigation of the formation of a thermoset wheat gluten network in a model system relevant for bread making

Although the impact of heat on molecular properties of wheat gluten is well understood, changes in its microstructure have rarely been studied. Here, formation of the thermoset gluten network in a model system relevant for bread baking was studied with confocal laser scanning microscopy and protein network analysis. From 65 °C onwards, gluten converts from thick aligned protein strands in a highly branched and homogeneous network of small thin protein threads. Neither gliadin incorporation in the network nor application of aqualysin 1, the thermo-active serine peptidase from Thermus aquaticus which recently has been reported to hydrolyse gluten proteins in dough only at temperatures exceeding 80 °C, impacts on the gluten microstructure. As starch causes structure setting itself and thereby decreases protein mobility, molecular scale changes in the gluten network at temperatures exceeding 80 °C brought about by aqualysin 1 do not impact its microstructure.     

Microstructural and dynamic oscillatory aspects of yogurt as influenced by hydrolysed guar gum

The objective of the current study was to explore the influence of crude, purified and hydrolysed guar gum at low and high concentrations on gel structure stability of yogurts at 4 °C throughout 21 days of storage. The rheology and microstructure of yogurts had been perceived by Bohlin rheometer (Malvern Instrument, UK) and cryo‐scanning electron microscopy (Cryo‐SEM), respectively. The viscoelastic behaviour of yogurts characterised at 25 °C exhibited weak gel structural properties (G? > G?). Cryo‐SEM had been used for microstructural image processing. The acidic and enzyme hydrolysed guar gum at low (0.3%) and high (1%) concentrations led to yogurt gels with fine‐stranded microstructure, while gelation with crude, purified and basic hydrolysed guar gum demonstrated diverse behaviour of depletion flocculation, visually observed even at low concentrations of crude guar gum (0.1%) as a thin layer of phase‐separated material. Conclusively, guar gum in its hydrolytic form enhances better structural and rheological properties.     

The Effect of Fermentation Temperature on the Microstructure,Physicochemical and Rheological Properties of Probiotic Buffalo Yoghurt

The properties of buffalo and bovine milk differ and the procedures developed to make bovine yoghurt may require optimisation for the production of buffalo yoghurt. This study aimed to apply cryo-scanning electron microscopy and confocal laser scanning microscopy to determine the optimal temperature for processing buffalo yoghurt. Milk was fermented at three different temperatures (37, 40 and 43 °C), stored for 28 days and the yoghurt microstructure, physicochemical and rheological properties assessed. Yoghurt fermented at 37 °C had a compact microstructure and the probiotic Lactobacillus acidophilus La-5 was more viable on storage. In contrast, yoghurt produced from a faster fermentation at 43 °C was firmer with a more porous microstructure that exhibited a higher degree of syneresis. The rheological properties during storage including the thixotropy, consistency coefficient and flow behaviour index were not significantly affected by temperature nor were the concentration of lactose, ionic calcium or titratable acidity. This study shows how changes to processing can be used to alter the microstructure of buffalo products and suggests that a decrease in fermentation temperature could be used to improve the quality of buffalo yoghurt.     

Effect of γ-polyglutamate on the rheological properties and microstructure of tofu

The effects of γ-polyglutamates with different concentrations (0.1%, 0.15%, 0.2%) and molecular weights (high, medium, low) addition on the rheological properties, microstructure, and syneresis of tofu were studied. The addition of γ-polyglutamate increased the gelation time, and decreased the storage modulus (G′) and the loss modulus (G″) of tofu. The molecular weight and concentration of γ-polyglutamate effectively changed the rheological properties of tofu. The network of tofu without γ-polyglutamate addition was constructed by fine strands in a dense arrangement as seen by using scanning electron microscope. However, the addition of γ-polyglutamate reduced the thickness of the strands in tofu network. Tofu syneresis was also reduced by the addition of γ-polyglutamate. Increase the concentration of γ-polyglutamate significantly decreased the syneresis of tofu. This trend was more evident on the tofu with high molecular weight γ-polyglutamate.     

Polymorphism, microstructure and rheology of butter. Effects of cream heat treatment

The effect of cream heat treatment prior to butter manufacturing, fluctuating temperatures during storage and presence of fat globules vs. no fat globules was examined in laboratory scale produced butter. X-ray diffraction and differential scanning calorimetry was used to study crystallization behaviour and nuclear magnetic resonance to measure solid fat content and water droplet size distribution. Furthermore, the crystal structure was linked to the rheological properties and microstructure of the butter using confocal laser scanning microscopy. Butter produced from non-matured cream mainly formed α- and β'-crystals with minor traces of β-crystals. Maturing of the cream caused a transition from α- to β'- and β-form. The rheological behaviour of slow cooled butter deviated from the matured ones by having a lower elastic modulus, caused by a weaker crystal network. Presence of fat globules did not affect the rheological properties significantly.     

Gel‐like emulsions prepared with zein nanoparticles produced through phase separation from acetic acid solutions

In this article, we report the microstructure and rheological property of Pickering emulsions stabilised with zein nanoparticles prepared through phase separation from acetic acid solution. The fresh emulsions showed liquid‐like behaviour and reasonable small droplet size. Interestingly, after 3 days of storage at 25° C, the emulsions changed into solid‐like state. The viscosity remarkably increased, and the storage modulus was much larger than the loss modulus. These results indicate the formation of the gel‐like network in emulsions. The droplet size also showed an obvious increase, while the big droplets could be disrupted into small ones in the presence of sodium dodecyl sulphate. The particle network in the continuous phase was seen in the confocal laser scanning microscopy. Therefore, it is suggested that the gel‐like network is formed by the flocculation of oil droplets and particle network in continuous, mainly through the hydrophobic interactions between the particles.     

Effect of autoclaving on the mechanical properties and structure of the giant squid hydrolysate-polysaccharide gel

Fish hydrolysates are an attractive option for preparing composite gel foods. However, not much is known regarding their gel properties. Here, we investigated the effect of the autoclaving treatment (121°C, 10 min) on the microstructure and properties of the giant squid hydrolysate-konjac glucomannan–κ-carrageenan–locust bean gum gel (GSH-P gel). The nuclear magnetic resonance proton spin–spin relaxation time (T₂) measurements indicated that autoclaving led to stronger water-binding ability of the GSH-P gel. The rheological measurements indicated that autoclaving led to significantly higher viscoelastic modulus and gel strength. The results of confocal laser scanning microscopy and small-angle X-ray scattering indicated that the phase separation of polysaccharides and proteins/peptides was enhanced by autoclaving, and the polysaccharides swelled better, resulting in less, but ordered, network structures. Autoclaving had a similar, but insignificant, positive effect on the polysaccharide (P) gel. Thus, GSH seems to play an important role in the process of polysaccharide gelation. This study shows that autoclaving can alter the structure and improve the properties of the GSH-P gel.     

The microstructure and textural properties of Australian cream cheese with differing composition

Confocal laser scanning microscopy was used to compare the microstructure of six Australian commercial cream cheese products. The optimal conditions for cryo scanning electron microscopy (cryo SEM) analysis of cream cheese microstructure were also examined. These complementary techniques revealed a typical cream cheese microstructure of homogenised fat globules embedded in a non-continuous protein network. The association between fat and protein within the microstructure was influenced by product composition (fat:protein ratio, moisture content) and ingredients. The addition of emulsifier led to a softer product with distinct microstructure. Cryo SEM also revealed a “honeycomb”-like structure, which was interpreted as a eutectic artefact formed by the addition of gum(s). Product hardness and gel strength generally correlated with high fat, low moisture content and a compact microstructure. Overall, this study shows how product composition affects the microstructure, texture and rheological properties of cream cheese.     

Effect of mechanical and thermal treatments on the microstructure and rheological properties of carrot,broccoli and tomato dispersions

BACKGROUND: The food industry has shown an increased interest in the manufacture of healthier and more natural food products. By tailored processing fruit and vegetables can be used as structurants thus reducing artificial gums and stabilisers. The effect of different thermal and mechanical treatments, including high‐pressure homogenisation, on the microstructural and rheological properties of carrot, broccoli and tomato dispersions was studied. As part of the rheological characterisation small oscillatory deformation as well as shear flow measurements were performed. RESULTS: Carrot and broccoli showed a different behaviour from tomato under the conditions studied. Changing the order of thermal and mechanical treatment led to microstructures with different flow properties. The resulting microstructures differed in the manner of cell wall separation: either breaking across the cell walls or through the middle lamella. High‐pressure homogenisation decreased the viscosity of carrot and broccoli dispersions, while it increased the viscosity of tomato. Cryo‐scanning electron microscopy showed that the cell walls of carrot and broccoli remained as compact structures after homogenisation whereas tomato cell walls were considerably swollen. CONCLUSIONS: Based on the type of vegetable, the different processes applied led to microstructures with different rheological properties. This study shows that particle size distribution, morphology and phase volume are important parameters to explain the complex relationship between rheology and microstructure for these types of systems. Copyright © 2010 Society of Chemical Industry     

Rheological,microstructural and sensory characterization of low-fat and whole milk set yoghurt as influenced by inulin addition

The effect of inulin addition (0–4%) upon texture and microstructure of set yoghurt with different levels of fat (0.2%–3.5%) was investigated. A two-factor experimental design with four treatments was used for data analysis. Skimmed milk with various inulin and cream concentrations was standardized to 4% protein content, homogenized, heated to 92 °C and fermented at 42 °C until a pH of 4.6 was reached. The chemical composition, pH, consistency and microstructure properties of the yoghurts were analysed after 6 days of storage at 5 °C. The statistical analysis showed that inulin and fat significantly affected the rheological and sensory results. Higher yield stress, “firmness” and “creaminess” values were observed in yoghurt produced with higher inulin additions, whereas the pH value was not affected. A significant correlation was found between yield stress and sensory determined firmness (r = 0.91). The microstructure examined by confocal laser scanning microscopy (CLSM) was only slightly affected by the concentrations of inulin in the range studied, possibly due to weak protein interactions between the inulin and the milk protein network.     

Impact of sodium chloride on wheat flour dough for yeast‐leavened products. I. Rheological attributes

BACKGROUND: The rheological properties of wheat dough for yeast‐leavened products were tested at different levels of sodium chloride (NaCl) addition ranging from 0 to 40 g NaCl kg^?1 wheat flour. Rheological tests carried out to make this evaluation included (1) empirical rheological methods of the Farinograph, load extension and a dough stickiness test and (2) fundamental rheological methods of creep recovery and dynamic rheometry. Modifications to the gluten matrix microstructure by NaCl were examined by confocal laser‐scanning microscopy. RESULTS: Highly significant (P?0.001) differences due to NaCl addition could be determined in particular by the stickiness test as well as by examination of the creep test with the Burger model. Rheological changes measured in the creep test probably depend on protein charge shielding due to NaCl interaction, resulting in an improvement in gluten network formation. An increase in dough stickiness was measured when using NaCl. CONCLUSION: The present result for stickiness is contrary to the common subjective results. Therefore the theory proposed here for increased stickiness suggests that it is based on more non‐protein‐bound water in the dough system due to NaCl interaction and thus more viscous dough behaviour, which leads to higher stickiness as measured with the stickiness test. This may also suggest that the objectively measured ‘stickiness’ in this case does not properly indicate the subjectively measured stickiness it was designed to represent. Copyright © 2011 Society of Chemical Industry     

EFFECT OF ADDITION OF PEACH GUM ON PHYSICOCHEMICAL PROPERTIES OF GELATIN-BASED MICROCAPSULE

The physicochemical properties of the microcapsules, which were prepared by spray drying, were evaluated. Gelatin‐sucrose (Gel‐Suc) and gelatin‐peach gum‐sucrose (Gel‐PG‐Suc) were used as wall material, while vitamin A was used as core material. The microencapsulation efficiency of Gel‐Suc and Gel‐PG‐Suc microcapsules significantly varied from 94.77 to 96.70% (P < 0.05). By adding peach gum, the microcapsules exhibited a spherical shape with a smooth surface, as evidenced by scanning electron microscopy, and the volume average diameter (D_4,3) showed significant difference from 73.95 to 68.72 µm (P < 0.05). By employing Fourier transform infrared spectroscopy, it was shown that OH‐stretching vibrations of Gel‐PG‐Suc microcapsules shifted to a lower wave number than that of Gel‐Suc microcapsules (Gel‐Suc microcapsules absorption peak: 3403.80/cm, Gel‐PG‐Suc microcapsules absorption peak: 3395.23/cm), so H‐bonding was strengthened by adding peach gum. Corresponding to the band of C=O in —C=ONH—, Gel‐Suc and Gel‐PG‐Suc microcapsules were 1652.72/cm and 1646.31/cm, respectively. Differential scanning calorimetry results revealed that the glass transition temperature (T_g) increased significantly from 46.103 to 50.448C by the addition of peach gum (P < 0.05).     

Rheological,thermal and microstructural properties of whey protein isolate‐modified cassava starch mixed gels at different pH values

The objective of this study was to investigate the rheological, thermal and microstructural properties of whey protein isolate (WPI)‐hydroxypropylated cassava starch (HPCS) gels and WPI‐cross‐linked cassava starch (CLCS) gels at different pH values (5.75, 7.00 and 9.00). The rheological results showed that the WPI‐modified starch gels had greater storage modulus (G?) values than the WPI‐native cassava starch gels at pH 5.75 and 7.00. Differential scanning calorimetry curves suggested that the phase transition order of the WPI and modified starch changed as the pH increased. Scanning electron microscopy images showed that the addition of HPCS and CLCS contributed to the formation of a compact microstructure at pH 5.75 and 7.00. A comprehensive analysis showed that the gelling properties of the WPI‐modified starch were affected by the difference between the WPI denaturation temperature and modified starch gelatinisation temperature and by the granular properties of the modified starch during gelatinisation. These results may contribute to the application of WPI‐modified starch mixtures in food preparation.     

Impact of arabinoxylan with different molecular weight on the thermo‐mechanical,rheological, water mobility and microstructural characteristics of wheat dough

The aim of this study was to investigate the effects of arabinoxylan with different molecular weight on the wheat dough thermo‐mechanical, rheological, microstructural and water mobility properties. Arabinoxylan was extracted from wheat bran and hydrolysed by endo‐1,4‐β‐xylanase (EC 3.2.1.8 from Trichoderma reesei, 10 000 U g^?1) for 2 min (AX_M) and 10 min (AX_L), respectively. The addition of hydrolysed arabinoxylan AX_L increased the stability time, decreased the setback value of wheat dough and enhanced the values of storage modulus (G′) and loss modulus (G″), while unhydrolysed arabinoxylan (AX_H) reduced these values. Meanwhile, unhydrolysed arabinoxylan increased T₂ relaxation time while hydrolysed arabinoxylan AX_L decreased T₂₁ and T₂₂. Confocal laser scanning microscope (CLSM) results showed that the addition of hydrolysed arabinoxylan AX_L promoted the formation of a more compact and continuous protein network in wheat dough. These results revealed that compared with high molecular weight arabinoxylan, hydrolysed arabinoxylan could improve the rheological properties and processing properties of wheat dough by enhancing the interaction among water molecules, starch and gluten in wheat dough.     

The effect of butter grains on physical properties of butter-like emulsions

Milk fat exists as globules in its natural state in milk. The potential of using globular fat to modulate the rheological properties and crystallization behavior in butter-like emulsions was studied in the present work. We conducted a comparative study of butter-like emulsions, with a fat phase consisting of 0, 10, 25, 50, or 100% anhydrous milk fat (AMF), the remaining fat being butter grains, and all samples containing 20% water, to obtain systematic variation in the ratio of globular fat. All emulsions were studied over 4 wk of storage at 5°C. By combining small and large deformation rheology, we conducted a detailed characterization of the rheological behavior of butter-like emulsions. We applied differential scanning calorimetry to monitor thermal behavior, confocal laser scanning microscopy for microstructural analysis, and low-field pulsed nuclear magnetic resonance spectrometry to measure solid fat content. By combining these techniques, we determined that increasing the fraction of globular fat (by mixing with butter grains) decreases the hardness of butter-like emulsions up to an order of magnitude at d 1. However, no difference was observed in thermal behavior as a function of butter grain content, as all emulsions containing butter grains revealed 2 endothermal peaks corresponding to the high (32.7°C ± 0.6) and medium (14.6°C ± 0.1) melting fractions of fatty acids. In terms of microstructure, decreasing the amount of butter grains in the emulsions resulted in formation of a denser fat crystal network, corresponding to increased hardness. Moreover, microstructural analysis revealed that the presence of butter grains resulted in faster formation of a continuous fat crystal network compared with the 100% AMF sample, which was dominated by crystal clusters surrounded by liquid oil. During storage, hardness remained stable and no changes in thermal behavior were observed, despite an increase in solid fat content of up to 5%. After 28 d of storage, we observed no difference in either microstructural or rheological properties, indicating that formation of primary bonds occurs primarily within the first day of storage. The rheological behavior of butter-like emulsions is not determined solely by hardness, but also by stiffness related to secondary bonds within the fat crystal network. The complex rheological behavior of milk fat-based emulsions is better characterized using multiple parameters.     

Extensional flow,viscoelasticity and baking performance of gluten-free zein-starch doughs supplemented with hydrocolloids

Viscoelastic doughs of zein and starch were prepared at 40 °C, above the glass transition temperature of zein. The effects of hydrocolloid supplementation with hydroxypropyl methylcellulose (HPMC) or oat bran with a high content of β-glucan (28%) were investigated by dynamic measurements in shear, confocal laser scanning microscopy (CLSM) and Hyperbolic Contraction Flow. Zein-starch dough without hydrocolloids exhibited rapid age-related stiffening, believed to be caused by cross-links between peptide chains. A prolonged softness was attributed to doughs containing hydrocolloids, with the oat bran exhibiting the most pronounced reduction in age-related stiffening. Moreover, CLSM-images of dough microstructure revealed that a finer fibre network may be formed by increased shearing through an addition of viscosity-increasing hydrocolloids, a reduction in water content in the dough or the use of appropriate mixing equipment. The Hyperbolic Contraction Flow measurements showed that doughs containing hydrocolloids had high extensional viscosities and strain hardening, suggesting appropriate rheological properties for bread making. Zein-starch dough without hydrocolloids showed poor bread making performance while hydrocolloid additions significantly improved bread volume and height. Although the hydrocolloid supplemented doughs had similar extensional rheological properties and microstructures, a fine crumb structure was attributed only to bread containing HPMC, marking the importance of surface active components in the liquid-gas interface of dough bubble walls. Zein could not mimic the properties of gluten on its own, but hydrocolloids did positively affect the structural and rheological properties of zein, which yielded dough similar to wheat dough and bread with increased volume.     

Effect of medium molecular weight xanthan gum in rheology and stability of oil‐in‐water emulsion stabilized with legume proteins

Xanthan gum is a water‐soluble extracellular polysaccharide that has gained widespread commercial use because of its strong pseudoplasticity and tolerance to high ionic strength, which bring unique rheological properties to solutions. This study compares and evaluates the emulsifying properties of oil‐in‐water (30:70 v/v) emulsions stabilized with lupin and soya protein isolates and medium molecular weight xanthan gum. The protein was obtained by an isoelectric precipitation method and the polysaccharide was produced by Xanthomonas campestris ATCC 1395 in batch culture in a laboratory fermenter (LBG medium) without pH control. The addition of xanthan gum in the emulsion formulation enhances emulsion stability through the phenomenon of thermodynamic incompatibility with the legume protein, resulting in an increase of the adsorbed protein at the interface. The emulsion stability is also enhanced by a network structure built by the polysaccharide in the bulk phase. Copyright © 2005 Society of Chemical Industry     

Effect of ground corn steeping on starch properties

Pasting and thermal properties, and microstructure of starch from ground corn (GS) steeped at 52 °C in sulphurous acid solution (0.20%) at different steep times (8, 16, 24, 32 and 40 h) were investigated. The isolated starch obtained by wet milling was characterised by determining pasting properties in Rapid Visco Analyser (RVA), retrogradation enthalpy in differential scanning calorimetry (DSC), and microstructure of granules in light (LM) and scanning electron microscopy (SEM). Milling before steeping and long steep time caused reduction in peak, hot paste, cool paste, breakdown and setback viscosities. Steep time provoked an increase in the retrogradation temperatures (onset and peak), whereas no significant differences were found in retrogradation enthalpy. Microstructure characteristics analysed by SEM showed many cracks and cavities on surface in samples steeped 8 h, whereas samples steeped 40 h showed a great alteration and disruption of their inner layer. LM with polarised light confirmed that starches retained birefringence with several differences depending on the steep time. The use of ground kernels allowed a better steep solution–endosperm interaction, which caused changes in rheological, thermal and microstructural starch characteristics.     

RHEOLOGICAL PROPERTIES OF OIL-IN-WATER EMULSIONS FORMED WITH MILK PROTEIN CONCENTRATE

The rheology and the microstructure of emulsions stabilized with milk protein concentrate (MPC) were investigated using different MPC and oil concentrations. Steady‐shear tests showed that the emulsions were Newtonian at low oil concentrations (<20 wt%), whereas they were pseudoplastic and thixotropic at high oil concentrations (>20 wt%). Dynamic rheological measurements demonstrated that these emulsions behaved as weak gels, as indicated by relatively high values of G′ to G″. The rheological behavior of these emulsions was consistent with bridging flocculation mechanisms, whereby more than one droplet is shared by adsorbed casein aggregates/micelles. This was confirmed by confocal microscopy, which showed that these emulsions were aggregated.     

ELECTROSTATIC EFFECTS ON PHYSICAL PROPERTIES OF PARTICULATE WHEY PROTEIN ISOLATE GELS

Physical properties of particulate whey protein isolate gels formed under varying electrostatic conditions were investigated using large strain rheological and microstructural techniques. The two treatment ranges evaluated were adjusting pH (5.2‐5.8) with no added NaCl and adjusting the NaCl (0.2‐0.6 M) at pH 7. Gels (10% protein w/v) were formed by heating at 80C for 30 min. The large strain properties of fracture strain (γ_f), fracture stress (σ_f), and a measure of strain hardening (R_0.3) were determined using a torsion method. Gel microstructure was evaluated using scanning electron microscopy (SEM) and gel permeability (B_gel). Overlaying σ_f and γ_f curves for pH and NaCl treatments demonstrated an overlap where gels of equal σ_f and γ_f could be formed by adjusting pH or NaCl concentration. The high fracture stress (σ_f～ 23 kPa and γ_f～ 1.86) pair conditions were pH 5.47 and 0.25 M NaCl, pH 7.0. The low fracture stress (σ_f～ 13 kPa and γ_f～ 1.90) pair conditions were pH 5.68 and 0.6 M NaCl, pH 7.0. The 0.25 M NaCl, pH 7 treatment demonstrated higher R_0.3 values than the pH 5.47 treatment. When the sulfhydryl blocker n‐ethylmaleimide was added at 2 mM to the 0.25 M NaCl, pH 7 gel treatment, its rheological behavior was NSD (p>0.05) to the pH 5.47 gel treatment, indicating disulfide bond formation regulated strain hardening. Altering surface charge or counterions, and disulfide bonding, was required to produce gels with similar large strain rheological properties. An increase in gel permeability coincided with an increase in pore size as observed by SEM, independent of rheological properties. This demonstrated that at the length scales investigated, microstructure was not linked to changes in large strain rheological properties.