Breadmaking Quality of Selected Durum Wheat Genotypes and Its Relationship with High Molecular Weight Glutenin Subunits Allelic Variation and Gluten Protein Polymeric Composition

Twenty‐seven durum wheat genotypes originating from different geographical areas, all expressing LMW‐2 at Glu‐B3, and five bread wheats were evaluated for flour mixing properties, dough physical characteristics, and baking performance. Gluten polymeric composition was studied using size‐exclusion HPLC of unreduced flour protein extracts. As a group, durum wheats had poorer baking quality than bread wheats in spite of higher protein and total polymer concentrations. Durum wheats exhibited weaker gluten characteristics, which could generally be attributed to a reduced proportion of SDS‐unextractable polymer, and produced less extensible doughs than did bread wheats. However, substantial variation in breadmaking quality attributes was observed among durum genotypes. Better baking performance was generally associated with greater dough extensibility and protein content, but not with gluten strength related parameters. Extensibility did not correlate with gluten strength or SEHPLC parameters. Genotypes expressing high molecular weight glutenin subunits (HMW‐GS) 6+8 exhibited better overall breadmaking quality compared with those expressing HMW‐GS 7+8 or 20. Whereas differences between genotypes expressing HMW‐GS 6+8 and those carrying HMW‐GS 7+8 could only be attributed to variations in extensibility, the generally inferior baking performance of the HMW‐GS 20 group relative to the HMW‐GS 6+8 group could be attributed to both weaker and less extensible gluten characteristics.     

Wheat Gluten Polymer Structures: The Impact of Genotype,Environment, and Processing on Their Functionality in Various Applications

For a number of applications, gluten protein polymer structures are of the highest importance in determining end‐use properties. The present article focuses on gluten protein structures in the wheat grain, genotype‐ and environment‐related changes, protein structures in various applications, and their impact on quality. Protein structures in mature wheat grain or flour are strongly related to end‐use properties, although influenced by genetic and environment interactions. Nitrogen availability during wheat development and genetically determined plant development rhythm are the most important parameters determining the gluten protein polymer structure, although temperature during plant development interacts with the impact of the mentioned parameters. Glutenin subunits are the main proteins incorporated in the gluten protein polymer in extracted wheat flour. During dough mixing, gliadins are also incorporated through disulfide‐sulfhydryl exchange reactions. Gluten protein polymer size and complexity in the mature grain and changes during dough formation are important for breadmaking quality. When using the gluten proteins to produce plastics, additional proteins are incorporated in the polymer through disulfide‐sulfhydryl exchange, sulfhydryl oxidation, β‐eliminations with lanthionine formation, and isopeptide formation. In promising materials, the protein polymer structure is changed toward β‐sheet structures of both intermolecular and extended type and a hexagonal close‐packed structure is found. Increased understanding of gluten protein polymer structures is extremely important to improve functionality and end‐use quality of wheat‐ and gluten‐based products.     

Improvement of Sorghum-Wheat Composite Dough Rheological Properties and Breadmaking Quality Through Zein Addition

Addition of sorghum flour to wheat flour produces marked negative effects on rheological properties of dough and loaf volume. Although there are notable differences in the chemical composition of sorghum proteins (kafirins) compared with wheat gluten that might imply poor functionality in breadmaking systems, a larger constraint may be the unavailability of kafirins due to encapsulation in protein bodies. In this study, zein, the analogous maize prolamin to kafirin, was used to determine the potential effects of protein-body-free prolamins on dough rheology and baking quality of wheat-sorghum composite flour. Mixograms run at 35°C (above the glass transition temperature of zein) were significantly (P < 0.01) improved with addition of zein. Mixogram peak heights increased while mixing time decreased uniformly with addition of zein. Dough extensibility studies showed an increase in maximum tensile stress, while baking studies showed an increase in loaf volume with increasing amounts of added zein. These data are supported by a previous study showing that, in a model system, zein mixed with starch can form viscoelastic networks, and suggest that kafirin, if made available, could contribute to dough formation.     

Formation of dityrosine cross-links during breadmaking

To establish its significance during commercial breadmaking, dityrosine formation was quantified in flours and doughs of six commercial wheat types at various stages of the Chorleywood Bread Process. Dityrosine was formed mainly during mixing and baking, at the levels of nmol/g dry weight. Good breadmaking flours tended to exhibit a higher dityrosine content in the final bread than low quality ones, but no relationship was found for dityrosine as a proportion of flour protein content, indicating that the latter was still a dominant factor in the analysis. There was no correlation between gluten yield of the six wheat types and their typical dityrosine concentrations, suggesting that dityrosine cross-links were not a determinant factor for gluten formation. Ascorbic acid was found to inhibit dityrosine formation during mixing and proving, and it has no significant effect on dityrosine in the final bread. Hydrogen peroxide promoted dityrosine formation, which suggests that a radical mechanism involving endogenous peroxidases might be responsible for dityrosine formation during breadmaking.     

Microbial Transglutaminase as a Tool to Restore the Functionality of Gluten from Insect‐Damaged Wheat

In some wheat‐growing countries, considerable quantities of commercial wheat are rendered unusable in standard baking because of preharvest damage of the grain by protease‐injecting bugs. In the present study, we studied the ability of transglutaminase (TG) treatment of damaged wheat flour to return the functionality of the gluten network. To confirm the TG cross‐linking, the degree of protein hydrolysis, the amount of free thiol groups, and the electrophoresis properties of glutenin subunits were determined. The effectiveness of the TG treatment on insect‐damaged wheat was analyzed by measuring the dough mixing behavior and the gluten quality. A decrease in the degree of hydrolysis (or free amino groups), a reduction in thiol group concentration, and a decrease of extractable high molecular weight glutenin subunits (HMW‐GS) (measured by high‐performance capillary electrophoresis) confirmed the protein cross‐linking catalyzed by TG, the simultaneous formation of disulfide bonds by the proximity of the cross‐linked polypeptide chains, and the formation of aggregates of high molecular weight. The TG treatment of the damaged wheat flour led to a recovery of the consistograph parameters and gluten index value, and the covalent nature of the bonds ensured the stability of the protein changes.     

Differential recovery of lupin proteins from the gluten matrix in lupin-wheat bread as revealed by mass spectrometry and two-dimensional electrophoresis

Bread made from a mixture of wheat and lupin flour possesses a number of health benefits. The addition of lupin flour to wheat flour during breadmaking has major effects on bread properties. The present study investigated the lupin and wheat flour protein interactions during the breadmaking process including dough formation and baking by using proteomics research technologies including MS/MS to identify the proteins. Results revealed that qualitatively most proteins from both lupin and wheat flour remained unchanged after baking as per electrophoretic behavior, whereas some were incorporated into the bread gluten matrix and became unextractable. Most of the lupin α-conglutins could be readily extracted from the lupin-wheat bread even at low salt and nonreducing/nondenaturing extraction conditions. In contrast, most of the β-conglutins lost extractability, suggesting that they were trapped in the bread gluten matrix. The higher thermal stability of α-conglutins compared to β-conglutins is speculated to account for this difference.     

Einkorn Characterization for Bread and Cookie Production in Relation to Protein Subunit Composition

Twenty-four einkorns were evaluated for agronomic traits in Italy and in Germany in replicated plot trials. After dehulling and milling, the harvested kernels, flour protein content, sedimentation volume, falling number, carotenoid, and dry gluten content were determined. Farinograph profiles were obtained with a farinograph and baking and cookie quality were evaluated with standard microtests. Significant differences in yield potential were observed between the two locations, with a three-fold increase in Germany as compared with Italy. One of the einkorn lines (ID529) had farinograph stability and degree of softening indices better than those of the control bread wheat. All the samples analyzed for breadmaking aptitude showed some degree of stickiness, but it was possible to handle the dough during the different steps of breadmaking. On average, cookies produced with einkorn flour were larger in diameter and thinner than those produced with soft wheat flour. The composition in α-, β- and γ-gliadins and in high molecular weight glutenin subunits was similar in all the lines. In contrast, the pattern exhibited in low molecular weight glutenin subunits correlated strictly with baking quality. In particular, the lines with bands arbitrarily designated a and b showed a high breadmaking potential, while the lines lacking these bands had an ample range of variability but, on average, a much lower baking potential. Our data point to a simple genetic control of the breadmaking aptitude and indicate einkorn not only as a promising source of specialty foods but also as an ideal species for genetic investigations on wheat quality.     

Effect of Physical States of Nonpolar Lipids on Rheology,Ultracentrifugation, and Microstructure of Wheat Flour Dough

In the previous study, we investigated effect of physical state of nonpolar lipids of gluten‐starch model dough. This experiment examined a real wheat flour dough system to assess the role of fat crystals in the breadmaking processes. These experiments were performed with a baking test and an investigation of wheat flour dough through rheological measurements (both large and small deformations), scanning electron microscopy, and ultracentrifugation. As a result, we found that the added oil was absorbed in the gluten structure, causing the aggregation of the gluten, which gave rise to more elastic behavior. In contrast, solid fat seemed to be distributed uniformly between the starch granules in the dough, reducing the friction between the starch granules and facilitating thin gluten gel layers. These properties lead to the lower G′ value and the increased viscous behavior, which yields an increase in loaf volume. In addition, the supposed mechanism behind the large loaf volume described in the previous study was that fat provides a uniform distribution of the dough components, and that the dough can thus expand easily, resulting in a larger loaf volume, which was supported in the wheat flour dough system. In conclusion, we found that thin, expandable gluten films and the uniform dispersion of gluten and starch granules in the dough are prerequisites for attaining better baking performance.     

Stress Relaxation Behavior of Wheat Dough,Gluten, and Gluten Protein Fractions

Relaxation behavior was measured for dough, gluten and gluten protein fractions obtained from the U.K. biscuitmaking flour, Riband, and the U.K. breadmaking flour, Hereward. The relaxation spectrum, in which relaxation times (τ) are related to polymer molecular size, for dough showed a broad molecular size distribution, with two relaxation processes: a major peak at short times and a second peak at times longer than 10 sec, which is thought to correspond to network structure, and which may be attributed to entanglements and physical cross‐links of polymers. Relaxation spectra of glutens were similar to those for the corresponding doughs from both flours. Hereward gluten clearly showed a much more pronounced second peak in relaxation spectrum and higher relaxation modulus than Riband gluten at the same water content. In the gluten protein fractions, gliadin and acetic acid soluble glutenin only showed the first relaxation process, but gel protein clearly showed both the first and second relaxation processes. The results show that the relaxation properties of dough depend on its gluten protein and that gel protein is responsible for the network structure for dough and gluten.     

Effect of Varying Protein Content and Glutenin-to-Gliadin Ratio on the Functional Properties of Wheat Dough

Gluten, starch, lipids, and water-soluble material were separated from seven wheat samples with a range of protein contents and breadmaking quality. The isolated glutens were further partitioned into gliadin- and gluteninrich fractions using pH precipitation. Protein content and glutenin-togliadin ratio were systematically altered by blending these fractions into the original flours in calculated amounts. Mixing properties, extension-tester parameters, and baking performance of composite flours were determined using small-scale techniques. Results of dough testing with blends of constant glutenin-to-gliadin ratio showed increases in the mixing time, mixograph peak resistance, maximum resistance to extension, extensibility, and loaf volume as the protein content increased. At constant protein content, increases in glutenin-to-gliadin ratio were associated with increases in mixing time, mixograph peak resistance, maximum resistance to extension, and loaf volume, and with decreases in extensibility. Thus, total protein content and glutenin-to-gliadin ratio independently affected dough and baking properties. The results have allowed the separation of the effects of flour protein quantity and composition on breadmaking properties.     

Comparison of Small and Large Deformation Rheological Properties of Wheat Dough and Gluten

The rheological properties of dough and gluten are important for end‐use quality of flour but there is a lack of knowledge of the relationships between fundamental and empirical tests and how they relate to flour composition and gluten quality. Dough and gluten from six breadmaking wheat qualities were subjected to a range of rheological tests. Fundamental (small‐deformation) rheological characterizations (dynamic oscillatory shear and creep recovery) were performed on gluten to avoid the nonlinear influence of the starch component, whereas large deformation tests were conducted on both dough and gluten. A number of variables from the various curves were considered and subjected to a principal component analysis (PCA) to get an overview of relationships between the various variables. The first component represented variability in protein quality, associated with elasticity and tenacity in large deformation (large positive loadings for resistance to extension and initial slope of dough and gluten extension curves recorded by the SMS/Kieffer dough and gluten extensibility rig, and the tenacity and strain hardening index of dough measured by the Dobraszczyk/Roberts dough inflation system), the elastic character of the hydrated gluten proteins (large positive loading for elastic modulus [G′], large negative loadings for tan δ and steady state compliance [J_e⁰]), the presence of high molecular weight glutenin subunits (HMW‐GS) 5+10 vs. 2+12, and a size distribution of glutenin polymers shifted toward the high‐end range. The second principal component was associated with flour protein content. Certain rheological data were influenced by protein content in addition to protein quality (area under dough extension curves and dough inflation curves [W]). The approach made it possible to bridge the gap between fundamental rheological properties, empirical measurements of physical properties, protein composition, and size distribution. The interpretation of this study gave indications of the molecular basis for differences in breadmaking performance.     

Association of Glutenin Subunit Composition and Dough Rheological Characteristics with Cookie Baking Properties of Soft Wheat Cultivars

The effect of flour type and dough rheology on cookie development during baking was investigated using seven different soft winter wheat cultivars. Electrophoresis was used to determine the hydrolyzing effects of a commercial protease enzyme on gluten protein and to evaluate the relationships between protein composition and baking characteristics. The SDS‐PAGE technique differentiated flour cultivars based on the glutenin subunits pattern. Electrophoresis result showed that the protease degraded the glutenin subunits of flour gluten. Extensional viscosities of cookie dough at all three crosshead speeds were able to discriminate flour cultivar and correlated strongly and negatively to baking performance (P < 0.0001). The cookie doughs exhibited extensional strain hardening behavior and those values significantly correlated to baking characteristics. Of all rheological measurements calculated, dough consistency index exhibited the strongest correlation coefficient with baking parameters. The degradation effects of the protease enzyme resulted in more pronounced improvements on baking characteristics compared with dough rheological properties. Stepwise multiple regression showed that the dough consistency index, the presence or absence of the fourth (44 kDa) subunit in LMW‐GS and the fifth subunit (71 kDa) subunit in HMW‐GS were predominant parameters in predicting cookie baking properties.     

小麦粉面团形成过程水分状态及其比例变化

为揭示小麦粉面团形成过程水分状态和比例、面团结构的变化,以及这种变化与粉质仪和拉伸仪表征的质量特性之间的关系;认识面团形成过程表征筋力强弱的物质基础和变化机理。选用中筋(宁春4号)和强筋(师栾02-1)小麦品种为试验材料,利用低场核磁共振技术测定粉质仪和面过程、拉伸仪醒发拉伸过程不同时间点面团水分状态和比例的变化;利用红外显微成像技术分析面团形成过程不同取样点蛋白质和淀粉的分布及结构变化。结果表明,面粉原料中主要为弱结合水。面粉在粉质仪加水搅拌形成面团后,水分状态和比例发生显著变化,面团中的水可以分为强结合水(T_(21))、弱结合水(T_(22))和自由水(T_(23))。面团搅拌形成过程中,中筋小麦品种宁春4号面团中的强结合水比例显著降低;师栾02-1的强结合水的弛豫时间在和面终点消失,弱结合水的弛豫时间显著延长,而自由水的比例显著增加(P0.05)。强筋小麦粉强结合水的保持时间较长。拉伸过程加盐和不加盐对同一取样点、同一种水分状态之间的水分弛豫时间和比例无显著影响;宁春4号自由水的弛豫时间在加盐和不加盐处理时都显著缩短(P0.05)。湿面筋含量高、筋力较强面团的蛋白质网络结构致密。粉质仪和面过程强结合水和弱结合水弛豫时间和比例的变化,与面筋含量和强度有关。该结论可为面制品加工过程和面工艺选择与优化等方面提供一定的理论参考。     

Protein Allelic Composition,Dough Rheology,and Baking Characteristics of Flour Mill Streams from Wheat Cultivars with Known and Varied Baking Qualities

Flour mill streams obtained by milling grain of 10 bread wheat cultivars grown in the Skopje region of Macedonia were analyzed for rheological and breadmaking quality characteristics and for composition of gliadins and HMW‐GS. The objective of this study was to examine the relationships between the composition of gluten proteins and breadmaking quality, as well as to determine the importance of gluten proteins for technological quality of flour mill streams. The grain was milled in an experimental mill according to a standardized milling procedure, with three break and three reduction passages. The addition of two vibratory finishers in the milling scheme enabled better separation of bran. A small‐scale baking method for evaluation of the breadmaking properties was developed, and electrophoretic methods including acid‐PAGE and SDS‐PAGE were used to determine the composition of the gluten proteins. There were significant differences in the degree of dough softening of individual and total flour fractions of the flour mill streams for cultivars with different alleles from six loci, for farinograph water absorption from seven loci, and for bread loaf volume and crumb quality score from six loci. The Glu‐1 quality scores for the wheat cultivars investigated were 3–9 and proved to be a useful indicator of breadmaking quality. The novel feature of the investigation related to the breadmaking potential of the flour mill streams compared with straight‐run flours.     

Relationships Between Gluten Rheological Properties and Hearth Loaf Characteristics

The rheological properties of fresh gluten in small amplitude oscillation in shear (SAOS) and creep recovery after short application of stress was related to the hearth breadbaking performance of wheat flours using the multivariate statistics partial least squares (PLS) regression. The picture was completed by dough mixing and extensional properties, flour protein size distribution determined by SE‐HPLC, and high molecular weight glutenin subunit (HMW‐GS) composition. The sample set comprised 20 wheat cultivars grown at two different levels of nitrogen fertilizer in one location. Flours yielding stiffer and more elastic glutens, with higher elastic and viscous moduli (G′ and G″) and lower tan δ values in SAOS, gave doughs that were better able to retain their shape during proving and baking, resulting in breads of high form ratios. Creep recovery measurements after short application of stress showed that glutens from flours of good breadmaking quality had high relative elastic recovery. The nitrogen fertilizer level affected the protein size distribution by an increase in monomeric proteins (gliadins), which gave glutens of higher tan δ and flatter bread loaves (lower form ratio).     

Effect of ascorbic acid in dough: reaction of oxidized glutathione with reactive thiol groups of wheat glutelin

The reactions of oxidized glutathione generated from endogenous glutathione by the addition of ascorbic acid (AA) prior to dough mixing on free thiol groups of gluten proteins have been investigated. A small amount of (35)S-labeled glutathione was added as a tracer to identify the reaction products of GSSG and free protein thiols by radioactivity measurement. First, gluten was isolated from the dough, then the gliadins were extracted, and residual glutenin was partially hydrolyzed with thermolysin. After preseparation by gel permeation chromatography, the fractions with the highest radioactivity were separated by high-performance liquid chromatography. Radioactive peptides were identified, isolated, sequenced, and assigned to amino acid sequences of gluten protein components. The isolated peptides contained exclusively the cysteine residues C(b) and C(x) of low molecular weight subunits of glutenin, which are supposed to be highly reactive in forming intermolecular disulfide bonds. From these results it can be assumed that the cysteine residues C(b) and C(x) of the low molecular weight subunits of glutenin are at least partly present in the thiol form in flour. During dough mixing they are converted to protein-protein disulfides or glutathione-protein mixed disulfides by thiol/disulfide interchange reactions. Oxidized glutathione necessary for this reaction is generated from glutathione by the action of AA. These results are in accordance with the major hypothesis about the mechanism of action of AA.     

Role of Water in Pretzel Dough Development and Final Product Quality

The relationship between flour quality or processing conditions and pretzel quality has not been extensively investigated. The objective of this study was to elucidate the role of water in pretzel dough development and the consequent impact on pretzel integrity. Control pretzel and pretzels made with lower or higher levels of added water in the dough were produced under standard processing conditions at Reading Bakery Systems' pilot plant in Robesonia, PA. Dough samples were evaluated for their appearance, moisture content, and extensibility and were viewed under a microscope to evaluate the gluten network. Pretzels before and after the kiln were evaluated for moisture content, pasting properties, and hardness and were viewed under a microscope to evaluate the extent of starch gelatinization. The structural and functional attributes of dough and pretzels were significantly different for the three treatments. The degrees of gluten development during mixing and starch gelatinization during baking were influenced by the levels of water added and consequently influenced pretzel quality. Pretzels made using low‐water treatment were brittle due to a lack of gluten development in the dough and inadequate starch gelatinization during baking, while pretzels made using high water treatment were unacceptable due to extensive gelatinization and retrogradation of starch. Pretzel quality therefore appeared to be a function of appropriate gluten development and starch gelatinization in the product.     

Comparison of Quality Characteristics and Breadmaking Functionality of Hard Red Winter and Hard Red Spring Wheat

Various whole‐kernel, milling, flour, dough, and breadmaking quality parameters were compared between hard red winter (HRW) and hard red spring (HRS) wheat. From the 50 quality parameters evaluated, values of only nine quality characteristics were found to be similar for both classes. These were test weight, grain moisture content, kernel size, polyphenol oxidase content, average gluten index, insoluble polymeric protein (%), free nonpolar lipids, loaf volume potential, and mixograph tolerance. Some of the quality characteristics that had significantly higher levels in HRS than in HRW wheat samples included grain protein content, grain hardness, most milling and flour quality measurements, most dough physicochemical properties, and most baking characteristics. When HRW and HRS wheat samples were grouped to be within the same wheat protein content range (11.4–15.8%), the average value of many grain and breadmaking quality characteristics were similar for both wheat classes but significant differences still existed. Values that were higher for HRW wheat flour were color b*, free polar lipids content, falling number, and farinograph tolerance. Values that were higher for HRS wheat flour were geometric mean diameter, quantity of insoluble polymeric proteins and gliadins, mixograph mix time, alveograph configuration ratio, dough weight, crumb grain score, and SDS sedimentation volume. This research showed that the grain and flour quality of HRS wheat generally exceeds that of HRW wheat whether or not samples are grouped to include a similar protein content range.     

Correlation of Quality Parameters with the Baking Performance of Wheat Flours

The baking performance of a set of flours from 13 wheat cultivars was determined by means of two different microscale baking tests (10 g of flour each). In the micro‐rapid‐mix test the dough was mixed for a fixed time at a high speed, whereas the microbaking test used mixing to optimum dough consistency in a microfarinograph. Quality parameters such as sedimentation value, crude protein content, dough and gluten extension data, and microfarinograph data were also determined. Finally, quality‐related protein fractions (gliadins, glutenins, SDS‐soluble proteins, and glutenin macropolymer) were quantitated by extraction/HPLC methods with reversed‐phase and gel‐permeation columns. All quality parameters were correlated with the bread volumes of both baking tests. The results demonstrated that the microbaking test (adapted mixing time) was much more closely related to the quality parameters than the micro‐rapid‐mix test (fixed mixing time), which hardly showed any correlation. Among the standard quality parameters, only the crude protein content showed a medium correlation with the bread volume of the microbaking test (r = 0.71), whereas the contents of gliadins (r = 0.80), glutenins (r = 0.76), and glutenin macropolymer (r = 0.80) appeared to be suitable parameters to predict the baking performance of wheat flour. All other quality parameters were not or were only weakly correlated and unsuitable for predicting baking performance.     

Effect of the Coenzymes NAD(P)(H) in Straight‐Dough Breadmaking on Protein Properties and Loaf Volume

The nicotinamide adenine dinucleotide coenzymes [NAD(P)(H)] are strong redox agents naturally present in wheat flour, and are indispensable cofactors in many redox reactions. Hence, it is not inconceivable that they affect gluten cross‐linking during breadmaking. We investigated the effect of increasing concentrations of NAD(P)(H) on gluten cross‐linking, dough properties, and bread volume using two flours of different breadmaking quality. Separate addition of the four nicotinamide coenzymes did not significantly affect mixograph properties. While addition of NAD⁺ hardly affected bread volume, supplementation with NADP(H) and NADH significantly decreased loaf volumes of breads made using flour of high breadmaking quality. Wheat flour incubation with NAD(P)H under anaerobic conditions increased wheat flour thiol content, while NAD(P)⁺ increased the extractability in SDS‐containing medium of the protein of the strong breadmaking flour. Based on the results, it was hypothesized that at least three reactions, competing for NAD(P)(H), occur during breadmaking that determine the final effect on protein, dough, and loaf properties. Next to coenzyme hydrolysis, the experiments pointed to coenzyme oxidation and NAD(P)(H) dependent redox reactions affecting protein properties.