Swelling behavior and structural characteristics of wheat gluten polypeptide films

Wheat gluten films were subjected to controlled thermomechanical treatments to increase the percentage of aggregated sodium dodecyl sulfate (SDS)-insoluble gluten protein, the aggregation reaction being disulfide bonding. The rheological properties of the films were measured under immersion in water, where wheat gluten films are stable and show only slight swelling. The equilibrium swelling of the gluten films in water decreased with the increase of the percentage of SDS-insoluble protein aggregates, and the frequency the independent shear modulus increased sharply with increasing percentage of SDS-insoluble aggregates. Both findings confirm that disulfide bonding between gluten proteins is the predominant cross-linking reaction in the system. A relationship between shear modulus and aggregated protein compatible with a power law (of exponent 3) suggests the existence of a protein network at a molecular scale. However, the classical Flory-Rehner model failed to describe the relationship between the plateau modulus and the gluten volume fraction (a very drastic increase, compatible with a power law of an exponent of about 14). This result shows that gluten cannot be described as an entangled polymer network. The interpretation of both relationships is a network of mesoscale particles which in turn have a fractal inner structure (with a fractal dimension close to 3).     

Analysis of dough functionality of flours from transgenic wheat

The rheological properties of flours from five different lines of transgenic wheat that either express or over-express subunits 1Ax1 and 1Dx5 were analyzed by mixograph assays and SDS sedimentation tests. In one case, the over-expression of subunit 1Dx5 resulted in a ca. 2-fold increase in mixing time, associated with a significant improvement in dough strength, and a lower resistance breakdown, suggesting an important increase in dough stability. However, the flour failed to develop properly without mixing with control flour because the rate of mixing was insufficient to develop the dough, i.e., the flour was overstrong. In two wheat transgenic lines, the expression of 1Ax1 and 1Dx5 transgenes, associated with silencing of all the endogenous high-molecular-weight glutenin subunits, resulted in flours with lower mixing time, peak resistance and sedimentation volumes, suggesting a lower gluten strength.     

A Spin-Label Study on Lipids in Dough Formed at Various Concentrations of Oxygen

The effect of oxygen on wheat flour lipids during dough mixing was investigated by analysis of the lipid composition and by an ESR technique with a fatty acid spin-label (4,4’-dimethyl-oxazolidine-N-oxyl derivative of 5-ketostearic acid). Dough was prepared in the presence of the spin-label under an atmosphere of air, nitrogen, 95% nitrogen—5% oxygen or oxygen, and the gluten was obtained by washing out the starch. ESR spectra of the spin-label incorporated into the gluten showed decreases in the order parameter, rotational correlation time and activation energy for rotational viscosity with increasing atmospheric oxygen concentration. During dough mixing in oxygen, oxidation of lipids proceeded and bound lipids slightly decreased. These data indicate that modification of lipids by incorporated oxygen leads to an increase in their fluidity and to a decrease in their hydrophobic interaction with protein in dough.     

The structure and properties of gluten: an elastic protein from wheat grain

The wheat gluten proteins correspond to the major storage proteins that are deposited in the starchy endosperm cells of the developing grain. These form a continuous proteinaceous matrix in the cells of the mature dry grain and are brought together to form a continuous viscoelastic network when flour is mixed with water to form dough. These viscoelastic properties underpin the utilization of wheat to give bread and other processed foods. One group of gluten proteins, the HMM subunits of glutenin, is particularly important in conferring high levels of elasticity (i.e. dough strength). These proteins are present in HMM polymers that are stabilized by disulphide bonds and are considered to form the 'elastic backbone' of gluten. However, the glutamine-rich repetitive sequences that comprise the central parts of the HMM subunits also form extensive arrays of interchain hydrogen bonds that may contribute to the elastic properties via a 'loop and train' mechanism. Genetic engineering can be used to manipulate the amount and composition of the HMM subunits, leading to either increased dough strength or to more drastic changes in gluten structure and properties.     

Characteristics of Relationships Between Structure of Gluten Proteins and Dough Rheology – Influence of Dietary Fibres Studied by FT-Raman Spectroscopy

The aim of this research was to study which kind of conformational changes in gluten proteins were induced by addition of four dietary fibre (apple-cranberry, cacao, carob and oat) by using FT-Raman spectroscopy and to find relationships between conformational changes and rheological behaviour of bread dough in mixing and extensional tests. Structural studies showed that all fibres induced formation of β-like structures between two protein molecules (pseudo-β-sheets) with the band at 1616 cm^?1 in the Raman spectrum. According to Principal Component Analysis, the strongest dependence was between changes in gluten structure and two extensographic parameters (resistance to extension and extensibility). Resistance to extension was positively correlated with content of α-helix and pseudo-β-sheets, while a negative correlation was observed between the parameter and content of β-sheets and β-turns. Gauche-gauche-gauche conformation of disulphide bridges and ability of tyrosine residues to hydrogen bonds creation improved mixing properties as stability of dough.     

Mechanism of heat and shear mediated aggregation of wheat gluten protein upon mixing

Changes in wheat gluten network structure upon mixing were studied from the biochemical analyses of gluten/glycerol blends mixed at 100 rpm with increasing times (up to 30 min) and temperatures of regulation (40, 60, and 80 degrees C). Whereas mixing induced protein solubility loss, the reduction of disulfide bonds restored protein extractability. But disulfide bond reduction became less efficient in promoting gluten extractability as mixing severity increased. This feature is consistent with the formation of a three-dimensional protein network stabilized by the formation of an increasing number of interchain disulfide bonds. Mixing induced a transient increase in free thiol groups while total thiol-equivalent groups dropped continuously. The changes were attributed to a shear-mediated scission of gluten disulfide bonds followed by oxidation of the thiyl radical moieties. Upon mixing, gluten solubility loss showed an Arrhenius-type temperature dependence with activation energy of 33.7 kJ.mol-1 instead of the more than 100 kJ.mol-1 reported for heat-induced gluten protein solubility loss. To explain this discrepancy, we postulated that during mixing, the disulfide interchange reactions are mediated by thiyl radicals in place of free thiol groups. A general model accounting for shear and temperature effects on gluten network structure is proposed.     

Application of xyloglucan to improve the gluten membrane on breadmaking

Effects of xyloglucan (XG) on the physical properties of dough and bread quality were studied. XG was fractionated to water-soluble (WS-XG) by enzymatic hydrolysis with cellulase, and the four kinds of WS-XG: WS-XG-A (average degree of polymerization; 17), WS-XG-B (32), WS-XG-C (78) and WS-XG-D (223) were obtained. XG without enzymatic hydrolysis was termed water-insoluble xyloglucan (WI-XG). Additions of WS-XG-A (3%), WS-XG-D (1–5%) and WI-XG (3%) increased the stability of the dough and improved the loaf and softness of bread samples. Especially, the WS-XG-D (1–5%) significantly improved the various factors of the final products, such as loaf volume, storage properties and good appearances with fine distribution of small size gas cells, and its addition of low level (1%) still showed improving effects, as compared with other additives. The more viscous gluten matrix could be observed in the mixed doughs with WS-XG-D, than the control sample without XG. WS-XG-D increased the water activity of the dough, and therefore the gluten matrix of dough became strong and uniform. Since the WS-XG-D had the higher degree of polymerization, it might be polymerized during mixing or fermentation, followed by the formation of new insoluble-XG after baking. Appropriate amount of the new WI-XG formed from WS-XG-D was considered to improve the storage properties with higher water holding property than WS-XG-D alone.     

Coexpression of the High Molecular Weight Glutenin Subunit 1Ax1 and Puroindoline Improves Dough Mixing Properties in Durum Wheat (Triticum turgidum L. ssp. durum)

Wheat end-use quality mainly derives from two interrelated characteristics: the compositions of gluten proteins and grain hardness. The composition of gluten proteins determines dough rheological properties and thus confers the unique viscoelastic property on dough. One group of gluten proteins, high molecular weight glutenin subunits (HMW-GS), plays an important role in dough functional properties. On the other hand, grain hardness, which influences the milling process of flour, is controlled by Puroindoline a (Pina) and Puroindoline b (Pinb) genes. However, little is known about the combined effects of HMW-GS and PINs on dough functional properties. In this study, we crossed a Pina-expressing transgenic line with a 1Ax1-expressing line of durum wheat and screened out lines coexpressing 1Ax1 and Pina or lines expressing either 1Ax1 or Pina. Dough mixing analysis of these lines demonstrated that expression of 1Ax1 improved both dough strength and over-mixing tolerance, while expression of PINA detrimentally affected the dough resistance to extension. In lines coexpressing 1Ax1 and Pina, faster hydration of flour during mixing was observed possibly due to the lower water absorption and damaged starch caused by PINA expression. In addition, expression of 1Ax1 appeared to compensate the detrimental effect of PINA on dough resistance to extension. Consequently, coexpression of 1Ax1 and PINA in durum wheat had combined effects on dough mixing behaviors with a better dough strength and resistance to extension than those from lines expressing either 1Ax1 or Pina. The results in our study suggest that simultaneous modulation of dough strength and grain hardness in durum wheat could significantly improve its breadmaking quality and may not even impair its pastamaking potential. Therefore, coexpression of 1Ax1 and PINA in durum wheat has useful implications for breeding durum wheat with dual functionality (for pasta and bread) and may improve the economic values of durum wheat.     

Effects of High-voltage Electric Field Treatment on the Water Activity of Bread

Treating bread dough by a high-voltage electric field (HVEF) during the first fermentation enabled the bread dough to retain water in the gluten fibers. The microstructure of the gluten fibers was still visible even with an HVEF treatment at 50 kV for 20 min, but could no longer be observed when the gluten was treated for more than 30 min. The crumb temperature of the HVEF treated bread during baking began to rise a few minutes sooner than that of the untreated bread, but the maximum temperature reached was the same in both cases: 99°C. The fact that the water activity of the HVEF-treated bread was 0.987 ± 0.0056, being higher than that of the untreated bread by 0.011, is in good agreement with the growing tests of Rhizopus nigricans. Furthermore, a distinct decrease in the water loss of the baked gluten was observed after the HVEF treatment. These results suggest that the HVEF treatment increased the ability of the gluten fibers, rather than the starch granules, to absorb and retain water; the state of the remaining water is considered to have become immobile, so that migration of moisture to the starch granules in the bread could be minimized.     

Reactivity of wheat gluten protein during mechanical mixing: radical and nucleophilic reactions for the addition of molecules on sulfur

Mechanical properties of gluten-based biomaterials, such as break stress, were known to be influenced by temperature and shear stresses applied during processing. It is well documented in literature that these processing parameters promoted wheat gluten protein aggregation. Exchange between disulfide bonds and thiol groups oxidation are the postulated mechanisms that lead to gluten protein solubility loss in sodium dodecyl sulfate buffers. Both nucleophilic and radical reactions were postulated to act during gluten aggregation. To graft molecules on gluten, a study was carried out to explore the reactivity of its thiol and disulfide groups during thermomechanical mixing. A range of reactants able to react via radical or nucleophilic pathways with thiol groups were synthesized. Reactivity between gluten and functions was quantified by gluten solubility measurements. This investigation and literature observations allowed proposal of a general gluten aggregation mechanism during mixing.     

土壤紧实度变化对小麦籽粒产量和品质的影响

以济南17(强筋品种)、烟农15(中筋品种)、鲁麦22(弱筋品种)为供试品种,设置人为碾压和不碾压2种处理,研究了土壤紧实度(以土壤容重表示)变化对不同类型小麦品种的籽粒产量和加工品质的影响。结果表明,随着土壤紧实度提高,3个品种的分蘖成穗率均显著降低,从而导致单位面积穗数和籽粒产量降低。3个品种相比较分蘖成穗率低的鲁麦22籽粒产量降幅最大。相关品质指标测定结果显示,提高土壤紧实度,对3个品种的蛋白质含量、湿面筋含量、沉淀值和吸水率均无显著影响,但济南17的面筋指数明显降低,面团断裂时间和面团稳定时间显著缩短,单位重量面粉烘焙所得的面包体积变小,而烟农15和鲁麦22受影响较小。其原因可能与土壤紧实度提高条件下济南17籽粒中谷蛋白／醇溶蛋白比例和谷蛋白大聚体含量降低有关。将济南17面团流变学特性年际间变化幅度与紧实度变化的处理效应相比较发现,土壤紧实度是影响强筋小麦品种品质性状稳定性的重要因素之一。     

小麦高分子量麦谷蛋白亚基序列及其结构与加工品质的关系

高分子量麦谷蛋白亚基（high molecular weight glutenin subunit,HMW-GS）是小麦种子贮藏蛋白的主要成分,其组成、含量和结构直接影响小麦面粉面筋的弹展性,决定着小麦的加工品质。本文主要对小麦HMW-GS的序列、结构和亚基之间组合形式做了详细的综述,并较系统地讨论了HMW-GS的结构和组成、特点等与面粉的加工品质之间的关系以及如何从定性和定量两方面来影响面粉的加工品质。     

Microstructures of bread dough and the effects of shortening on frozen dough

Three types of straight doughs different in combination of yeast and shortenings (RLS20, FTS20, and FTS80) were prepared, and the structure of the frozen doughs was examined under a microscope after staining protein or lipid droplets. Even after 2 months of frozen storage, distinct changes were not found in the gluten network of FTS80, although significant damages in the dough structures of FTS20 and RLS20 appeared after only one month of frozen storage. These results suggest that the gluten networks loosen and decrease in the water retention ability, and it may be concluded that the lipid is removed from the gluten protein due to the decrease in water in the continuous protein phase. The resulting product from the damage to the gluten matrix gave rise to fusion of lipid droplets and an increase in their size. Because of the difference in fatty acid composition, the lipids of shortening S80 are presumed to interact more strongly with gluten proteins and to keep the gluten matrix from damage in comparison with the lipids of shortening S20.     

Creating Novel Structures in Food Materials: The Role of Well-Defined Shear Flow

Structure formation in food materials is influenced by the ingredient properties and processing conditions. Until now, small structural elements, such as fibrils and crystals, have been formed using self-assembly, while processing was applied to create relatively large structures. The effect of self-assembly under flow is rarely studied for food materials, but it is widely studied for non-food systems. The use of well-defined flow, often simple shear, turned out to be essential to study and control the structure formation process in foods as well. This observation encouraged us to develop a number of different shearing devices that allowed processing of biopolymer systems under simple shear flow. This paper reviews our main findings. In the case of protein fibrillization, the shear rate was found to control the growth rate as well as the properties of the fibrils formed. In the case of dough processing, simple shear flow made the product more process tolerant and induced gluten migration. The use of shear flow for dense caseinate dispersions led to hierarchically structured and fibrous material. Based on the presented results, we conclude that introducing simple shear flow in food structuring processes can lead to a much broader range of structures, thereby better utilizing the full potential of food ingredients.     

Rheological and breadmaking properties of wheat samples infected with<Emphasis Type="Italic">Fusarium spp.</Emphasis>

Rheological and breadmaking properties of untreated and suboptimally stored wheat samples (grain moisture: 20%, temperature: 20°C) and also of wheat which was inoculated withFusarium spp. were investigated. The deoxynivalenol (DON) content of the stored and inoculated wheat samples ranged between 820–12,000 μg/kg. Gluten proteins were isolated with different extraction solutions and the fractions obtained were analysed by means of RP-HPLC. Microextension tests and micro-baking tests were used for the determination of dough properties (maximum resistance (MR) and extensibility (EX)) and bread volume, respectively. In spite of the extremely high DON concentrations of some wheat samples contaminated withFusarium spp. they showed only a slight decrease of the amount of gluten proteins. Extension tests of dough led to a slight decrease of MR, bread volumes stayed almost the same compared with the non-contaminated grain. The contamination of wheat withAspergillus andPenicillium led to a high decrease of gluten proteins, which resulted in an extremely decreased MR of the dough and a very low bread volume.     

Distribution of gluten proteins in bread wheat (Triticum aestivum) grain

Background and Aims

Gluten proteins are the major storage protein fraction in the mature wheat grain. They are restricted to the starchy endosperm, which forms white flour on milling, and interact during grain development to form large polymers which form a continuous proteinaceous network when flour is mixed with water to give dough. This network confers viscosity and elasticity to the dough, enabling the production of leavened products. The starchy endosperm is not a homogeneous tissue and quantitative and qualitative gradients exist for the major components: protein, starch and cell wall polysaccharides. Gradients in protein content and composition are the most evident and are of particular interest because of the major role played by the gluten proteins in determining grain processing quality.

Methods

Protein gradients in the starchy endosperm were investigated using antibodies for specific gluten protein types for immunolocalization in developing grains and for western blot analysis of protein extracts from flour fractions obtained by sequential abrasion (pearling) to prepare tissue layers.

Key Results

Differential patterns of distribution were found for the high-molecular-weight subunits of glutenin (HMW-GS) and γ-gliadins when compared with the low-molecular-weight subunits of glutenin (LMW-GS), ω- and α-gliadins. The first two types of gluten protein are more abundant in the inner endosperm layers and the latter more abundant in the subaleurone. Immunolocalization also showed that segregation of gluten proteins occurs both between and within protein bodies during protein deposition and may still be retained in the mature grain.

Conclusions

Quantitative and qualitative gradients in gluten protein composition are established during grain development. These gradients may be due to the origin of subaleurone cells, which unlike other starchy endosperm cells derive from the re-differentiation of aleurone cells, but could also result from the action of specific regulatory signals produced by the maternal tissue on specific domains of the gluten protein gene promoters.     

Rheological behaviour and physical properties of controlled-release gluten-based bioplastics

Bioplastics based on glycerol, water and wheat gluten have been manufactured in order to determine the effect that mechanical processing and further thermal treatments exert on different thermo-mechanical properties of the biomaterials obtained. An “active agent”, KCl was incorporated in these matrices to develop controlled-release formulations. Oscillatory shear, dynamic mechanical thermal analysis (DMTA), diffusion and water absorption tests were carried out in order to study the influence of the above-mentioned treatments on the physico–chemical characteristics and rheological behaviour of these bioplastic samples. Wheat gluten protein-based bioplastics studied in this work present a high ability for thermosetting modification, due to protein denaturation, which may favour the development of a wide variety of biomaterials. Bioplastic hygroscopic properties depend on plasticizer nature and processing procedure, and may be a key factor for industrial applications where water absorption is required. On the other hand, high water absorption and slow KCl release from bioplastic samples (both of them suitable properties in agricultural applications) may be obtained by adding citric acid to a given formulation, at selected processing conditions.     

Role of lipoxygenase in the determination of wheat grain quality

Analysis of the correlation between endogenous lipoxygenase activity and 15 wheat grain quality parameters in three bread wheat populations has shown that enzyme activity influences the weight of 1000 grains, dough deformation energy, dough tenacity, and mixing properties. The correlations between the enzyme activity and the basic quality parameters are negative at high activity levels. The optimum values of specific lipoxygenase activity at which all quality parameters studied have the maximum values range from 108.5 ± 1.2 to 126.4 ± 1.9. It has been found that the ability of lipoxygenase to strengthen gluten is related to the lowering of dough extensibility.     

Visualization of gluten and starch distributions in dough by fluorescence fingerprint imaging

A novel method combining imaging techniques and fluorescence fingerprint (FF) data measurement was developed to visualize the distributions of gluten and starch in dough without any preprocessing. Fluorescence images of thin sections of gluten, starch, and dough were acquired under 63 different combinations of excitation and emission wavelengths, resulting in a set of data consisting of the FF data for each pixel. Cosine similarity values between the FF of each pixel in the dough and those of gluten and starch were calculated. Each pixel was colored according to the cosine similarity value to obtain a pseudo-color image showing the distributions of gluten and starch. The dough sample was then fluorescently stained for gluten and starch. The stained image showed patterns similar to the pseudo-color FF image, validating the effectiveness of the FF imaging method. The method proved to be a powerful visualization tool, applicable in fields other than food technology.     

Visualization of Gluten and Starch Distributions in Dough by Fluorescence Fingerprint Imaging

A novel method combining imaging techniques and fluorescence fingerprint (FF) data measurement was developed to visualize the distributions of gluten and starch in dough without any preprocessing. Fluorescence images of thin sections of gluten, starch, and dough were acquired under 63 different combinations of excitation and emission wavelengths, resulting in a set of data consisting of the FF data for each pixel. Cosine similarity values between the FF of each pixel in the dough and those of gluten and starch were calculated. Each pixel was colored according to the cosine similarity value to obtain a pseudo-color image showing the distributions of gluten and starch. The dough sample was then fluorescently stained for gluten and starch. The stained image showed patterns similar to the pseudo-color FF image, validating the effectiveness of the FF imaging method. The method proved to be a powerful visualization tool, applicable in fields other than food technology.