Polymer Conformation Structure of Wheat Proteins and Gluten Subfractions Revealed by ATR‐FTIR

The polymer conformation structure of gluten extracted from a Polish wheat cultivar, Korweta, and gluten subfractions obtained from 2 U.K. breadmaking and biscuit flour cultivars, Hereward and Riband, was investigated using attenuated total reflectance Fourier transform infrared spectroscopy (ATR‐FTIR). The results showed the conformation of proteins varied between flour, hydrated flour, and hydrated gluten. The β‐sheet structure increased progressively from flour to hydrated flour and to hydrated gluten. In hydrated gluten protein fractions comprising gliadin, soluble glutenin, and gel protein, β‐sheet structure increased progressively from soluble gliadin and glutenin to gluten and gel protein; β‐sheet content was also greater in the gel protein from the breadmaking flour Hereward than the biscuit flour Riband.     

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.     

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.     

Relationship Between the Qualitative and Quantitative Compositions of Gluten Protein Types and Technological Properties of Synthetic Hexaploid Wheat Derived from Triticum durum and Aegilops tauschii

The contribution of the diploid wheat species Aegilops tauschii (Coss.) Schmall to the technological properties of bread wheat (Triticum aestivum L.) was previously studied by the investigation of synthetic hexaploids derived from tetraploid durum wheat (T. turgidum L.) and three diploid Ae. tauschii lines. The results indicated that bread volume, gluten index, SDS‐sedimentation volume, and maximum resistance of gluten were significantly influenced by the Ae. tauschii lines. To determine the relationship between technological properties and qualitative and quantitative compositions of gluten proteins, the flours of parental and synthetic lines were extracted using a modified Osborne fractionation. Gliadin and glutenin fractions were then characterized by reversed‐phase (RP) HPLC on C₈ silica gel. The HPLC patterns revealed typical differences between synthetic and parental lines. The gliadin patterns of three synthetic lines and the glutenin patterns of two synthetic lines were more similar to that of the diploid Ae. tauschii parents involved in the hybrids. In the glutenin pattern of one synthetic line, characteristics from both Ae. tauschii and the durum wheat parents were observed. The amount of total gliadin and gliadin types of the synthetic lines was mostly intermediate between those of the durum and Ae. tauschii parents. The amounts of total glutenin and glutenin types (HMW and LMW subunits) of the synthetic lines were generally higher than those of the parental lines, and the ratio of gliadins to glutenins was significantly decreased. High positive correlations were found between the amount of total glutenins, HMW, and LMW subunits and bread volume, maximum resistance and extension area of gluten, and SDS‐sedimentation volume. The ratio of gliadins to glutenin subunits had a strong negative influence on these properties. The protein content of the flours and the amount of total gluten proteins were not correlated with any of the technological properties. Results on the relationship between biochemical characteristics and the breadmaking properties indicated that wheat prebreeding would benefit from studies on protein types and quantification in the choice of parents. In addition, the potential of the diploid Ae. tauschii for improvement of breadmaking quality should be further exploited.     

Comparing Small‐Scale Testing Methods for Predicting Wheat Gluten Strength Across Environments

Gluten strength is the main factor determining the rheological and processing properties of wheat. Rapid, small‐scale tests that can indirectly predict gluten strength are extremely important for wheat‐breeding selection, particularly when using pedigree methodology. The efficiency and reliability of three small‐scale tests (SDS sedimentation volume [SDSS], swelling index of glutenin [SIG], and lactic acid retention capacity [LARC]) across three environments (E1, no stress; E2, drought stress; and E3, heat stress) were evaluated by using 15 common wheat and nine durum wheat cultivars. In the case of common wheat, SIG highlighted its advantage for predicting gluten strength, even under stress environments, compared with LARC and SDSS, whereas SDSS showed the best relationship with bread loaf volume. For durum wheat, SIG showed the best predicting value in E1 and E3; however, under drought stress, SDSS, SIG, and LARC all lost their good ability for predicting gluten strength in durum wheat, which needs further investigation. Also, the comparison between two mixograph parameters (mixograph peak time and mixograph peak integral) for predicting gluten strength and the suitability of testing SIG and LARC with whole meal (or semolina) instead of refined flour were also investigated.     

Does Fumigation of Durum Wheat and Semolina with Sulfuryl Fluoride Affect Quality of the Grain,Semolina, and Derived Spaghetti and Bread?

There is no information on the effect of sulfuryl fluoride (SF) on durum wheat technological properties and products made from fumigated durum wheat. Durum wheat and semolina were exposed to a range of SF applications under conditions that might be typically encountered in bulk storage facilities used in many countries. SF greatly reduced the germination percentage of fumigated durum wheat, with increasing impact under higher SF concentration, grain moisture content, and fumigation temperature. SF greatly reduced seed germination percentage, impacting more the higher the SF concentration. SF had little to no effect on grain test weight, 1,000‐grain weight, hardness, protein content, semolina ash content, and mixograph properties. At the highest SF concentration (31.25 mg/L for 48 h) there was a tendency for pasta cooking loss to be increased but still acceptable, and other pasta properties were largely unaffected. Fumigation with SF did not have any impact on the baking properties of a wholemeal durum flour–commercial flour mix. Therefore, SF is not recommended if the grains are to be used as seeds for agricultural production, but for the production of semolina, pasta, and bread, SF used under typical fumigation conditions has little to no impact on technological properties of durum wheat.     

New Insights Into the Role of Gluten on Durum Pasta Quality Using Reconstitution Method

The effects of varying the gluten composition at constant protein, protein content at constant composition, and glutenin‐to‐gliadin (glu/gli) ratio on durum semolina rheological properties and the quality of the spaghetti derived from these doughs was investigated using the reconstitution method. Reconstituted flours were built up from a common durum starch and water‐soluble fraction but with varying gluten types from a range of wheats at both 12 and 9% total protein. A 10‐g mixograph and microextensigraph properties were affected by the source of the gluten, which was related to glutenin composition and polymeric molecular weight distribution. Cooked pasta firmness was highly correlated to mixograph development time (MDDT). Furthermore, varying the protein content (9–20%) showed an increase in mixograph peak resistance (PR) with no effect on extensigraph Rmax. Pasta firmness increased and stickiness decreased with increasing protein content. In another experiment, the glutenin and gliadin fractions isolated from durum wheat were added to the respective base semolina to investigate the effect of varying the glu/gli ratio by 1.3–1.6 fold. Increasing the ratio increased MDDT but had no effect on PR and resistance breakdown. Variable effects were obtained for spaghetti firmness. The information obtained should prove useful to durum breeders by providing further evidence for the importance of protein to pasta quality.     

Starch Participation in Durum Dough Linear Viscoelastic Properties

The contribution of starch to dough rheological properties has been largely overshadowed by the role of gluten, receiving much less attention in comparison. The influence of starch granule surface properties on durum wheat dough linear viscoelasticity was investigated, and surface interactions between starch granules and gluten were assessed using a model system. Proportions of starch were substituted in dough on a volume basis with an inert filler (glass powder) with a similar particle size range. The doughs were subjected to dynamic and creep measurements. Dough linear viscoelastic properties were weakened on substitution of starch with glass powder at ≤50% substitution, inferring a reduction in adhesion at the matrix‐filler (starch and glass powder) interface with declining proportions of starch granules. Surface modification of starch granules or glass powder altered dough rheological properties, confirming the importance of starch granule surface characteristics and the nature of protein‐starch bonding on durum dough linear viscoelastic behavior.     

Antioxidants, free radicals, storage proteins, puroindolines, and proteolytic activities in bread wheat (Triticum aestivum) seeds during accelerated aging

Seeds of bread wheat were incubated at 40 degrees C and 100% relative humidity for 0, 3, 4, 6, and 10 days. The effects of accelerated aging on seed germinability and some biochemical properties of flour (carotenoid, free radical, and protein contents and proteolytic activity) and gluten (free radical content and flexibility) were investigated. Seed germinability decreased during aging, resulting in seed death after 10 days. A progressive decrease of carotenoid content, in particular, lutein, was observed, prolonging the incubation, whereas the free radical content increased in both flour and gluten. A degradation of soluble and storage proteins was found, associated with a marked increase of proteolytic activity and a loss of viscoelastic properties of gluten. On the contrary, puroindolines were quite resistant to the treatment. The results are discussed in comparison with those previously obtained during accelerated aging of durum wheat seeds.     

Study of Viscoelastic Properties of Wheat Kernels Using Compression Load Method

The method to measure hardness and other viscoelastic properties of intact wheat kernels is presented. Wheat with 9.3% moisture showed high elastic behavior compared with wheat tempered at 22.5% moisture that showed a plastic behavior. Load‐deformation curves showed that bread wheat behaves as a more plastic material than durum wheat, which is a more elastic material. Yield point of all the wheat samples was ≈18.5 N, independent of wheat type and moisture content. The height of the wheat kernel increased linearly, and the compression area increased exponentially, with increasing moisture content. The modulus of elasticity of wheat ranged from 99.2 MPa for 22.5% moisture content to 394.8 MPa for 9.3% moisture content. Young's modulus range for soft wheat such as Salamanca, Saturno, and Cortazar cultivars was 232.2–308.5 MPa, as compared with Rayón bread wheat at 321.5 MPa and the Altar, Sofía, and Rafi cultivars of durum wheat that had elastic moduli of 438.7–485.8 MPa. The compression force and final stress decreased from 69.9 N and 40.1 MPa in soft wheat to 90 N and 78.9 MPa in durum, respectively. Total work range was 14.7 MPa/sec in soft wheat to 19.7 MPa/sec for durum wheat and, as expected, was higher in the durum and bread wheat than in soft wheat. The plastic part ranged from 2.4 MPa/sec in soft wheat to 0.6 MPa/sec in durum wheat.     

Qualitative Effect of Added Gluten on Dough Properties and Quality of Chinese Steamed Bread

Glutens isolated from 15 soft red winter (SRW) wheat flours were added into a SRW wheat flour to obtain protein levels of 9.6 and 11.3% for determination of the qualitative effect of added gluten on the dough properties and quality of northern‐style Chinese steamed bread (CSB). Sodium dodecyl sulfate sedimentation (SDSS) volume of the gluten source flour exhibited positive relationships with mixograph absorption, midline peak time (MPT), and midline peak value (MPV) of the gluten‐added flours and with surface smoothness, crumb structure, and total score of CSB prepared from the gluten‐added flours regardless of protein content. Positive correlations were also observed between SDSS volume of the gluten source flour and specific volume and stress relaxation score of CSB prepared from the gluten‐added flours of 11.3% protein. The increase in protein content from 9.6 to 11.3% by gluten addition raised mixograph absorption, MPT, and MPV but had no apparent effect on resistance breakdown, dough maximum force for extension, and extensibility, and it increased CSB specific volume and crumb structure score without affecting surface smoothness, stress relaxation, and total score. Mixograph parameters exhibited significant relationships with CSB total score, indicating that they could be effective predictors of the CSB‐making quality of flours.     

Effects of Wheat Cultivar and Nitrogen Application on Storage Protein Composition and Breadmaking Quality

Influences of cultivar and nitrogen application on protein concentration and composition, and amount and size‐distribution of different protein components, were investigated in 10 spring wheat cultivars (Triticum aestivum L.) with widely varying gluten strength, grown under four nitrogen fertilizer conditions. The results showed that cultivar differences in gluten strength were determined by storage protein composition, differences in total amount of HMW glutenin subunits, the glutenin‐to‐gliadin ratio, and the relationship between SDS‐soluble and SDS‐insoluble protein polymers. Negative correlations were found between protein parameters related to gluten strength and bread volume. No cultivar stability for gluten strength in relation to differences in nitrogen application was found. Thus, the gluten strength was influenced by the nitrogen application in all the investigated cultivars. Increased nitrogen supply correlated significantly to an increase in all protein components containing gliadins and glutenins, but not to those containing albumins and globulins. The increase in protein components containing gliadins and glutenins correlated significantly with an increase in protein concentration and bread volume.     

Protein and Quality Characterization of Complete and Partial Near‐Isogenic Lines of Waxy Wheat

The objective of this study was to evaluate protein composition and its effects on flour quality and physical dough test parameters using waxy wheat near‐isogenic lines. Partial waxy (single and double nulls) and waxy (null at all three waxy loci, Wx‐A1, Wx‐B1, and Wx‐D1) lines of N11 set (bread wheat) and Svevo (durum) were investigated. For protein composition, waxy wheats in this study had relatively lower albumins‐globulins than the hard winter wheat control. In the bread wheats (N11), dough strength as measured by mixograph peak dough development time (MDDT) (r = 0.75) and maximum resistance (R_max) (r = 0.70) was significantly correlated with unextractable polymeric protein (UPP), whereas in durum wheats, moderate correlation was observed (r = 0.73 and 0.59, respectively). This may be due to the presence of high molecular weight glutenin subunits (HMW‐GS) Dx2+Dy12 at the Glu‐D1 locus instead of Dx5+Dy10, which are associated with dough strength. Significant correlation of initial loaf volume (ILV) to flour polymeric protein (FPP) (r = 0.75) and flour protein (FP) (r = 0.63) was found in bread wheats, whereas in durum wheats, a weak correlation of ILV was observed with FP (r = 0.09) and FPP (r =0.51). Significant correlation of ILV with FPP in bread wheats and with % polymeric protein (PPP) (r = 0.75) in durum lines indicates that this aspect of end‐use functionality is influenced by FPP and PPP, respectively, in these waxy wheat lines. High ILV was observed with 100% waxy wheat flour alone and was not affected by 50% blending with bread wheat flour. However, dark color and poor crumb structure was observed with 100% waxy flour, which was unacceptable to consumers. As the amylopectin content of the starch increases, loaf expansion increases but the crumb structure becomes increasingly unstable and collapses.     

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.     

Phytosiderophore release does not relate well with zinc efficiency in different bread wheat genotypes

Using six bread wheat genotypes (Triticum aesttvum L. cvs. Dagdas‐94, Gerek‐79, BDME‐10, SBVD 1–21, SBVD 2–22 and Partizanka Niska) and one durum wheat genotype (Triticum durum L. cv. Kunduru‐1149) experiments were carried out to study the relationship between the rate of phytosiderophore release and susceptibility of genotypes to zinc (Zn) deficiency during 15 days of growth in nutrient solution with (1 μM Zn) and without Zn supply. Among the genotypes, Dagdas‐94 and Gerek‐79 are Zn efficient, while the others are highly susceptible to Zn deficiency, when grown on severely Zn deficient calcareous soils in Turkey. Similar to the field observations, visual Zn deficiency symptoms, such as whitish‐brown lesions on leaf blades occurred first and severely in durum wheat Kunduru‐1149 and bread wheats Partizanka Niska, BDME‐10, SBVD 1–21 and SBVD 2–22. Visual Zn deficiency symptoms were less severe in the bread wheats Gerek‐79 and particularly Dagdas‐94. These genotypic differences in susceptibility to Zn deficiency were not related to the concentrations of Zn in shoots or roots. All bread wheat genotypes contained similar Zn concentration in the dry matter. In all genotypes supplied adequately with Zn, the rate of phytosiderophore release was very low and did not exceed 0.5 μmol/48 plants/ 3 h. However, under Zn deficiency the release of phytosiderophores increased in all bread wheat genotypes, but not in the durum wheat genotype. The corresponding rates of phytosiderophore release in Zn deficient durum wheat genotype were 1.2 umol and in Zn deficient bread wheat genotypes ranged between 8.6 μmol for Partizanka Niska to 17.4 umol for SBVD 2–22. In Dagdas‐94, the most Zn efficient genotype, the highest rate of phytosiderophore release was 14.8 umol. The results indicate that the release rate of phytosiderophores does not relate well with the susceptibility of bread wheat genotypes to Zn deficiency. Root uptake and root‐to‐shoot transport of Zn and particularly internal utilization of Zn may be more important mechanisms involved in expression of Zn efficiency in bread wheat genotypes than release of phytosiderophores.     

Textural Comparisons of Gluten‐Free and Wheat‐Based Doughs,Batters, and Breads

Studies were conducted with two newly developed gluten‐free bread recipes. One was based on corn starch (relative amount 54), brown rice (25), soya (12.5), and buckwheat flour (8.5), while the other contained brown rice flour (50), skim milk powder (37.5), whole egg (30), potato (25), and corn starch (12.5), and soya flour (12.5). The hydrocolloids used were xanthan gum (1.25) and xanthan (0.9) plus konjac gum (1.5), respectively. Wheat bread and gluten‐free bread made from commercial flour mix were included for comparison. Baking tests showed that wheat and the bread made from the commercial flour mix yielded significantly higher loaf volumes (P < 0.01). All the gluten‐free breads were brittle after two days of storage, detectable by the occurrence of fracture, and the decrease in springiness (P < 0.01), cohesiveness (P < 0.01), and resilience (P < 0.01) derived from texture profile analysis. However, these changes were generally less pronounced for the dairy‐based gluten‐free bread, indicating a better keeping quality. Confocal laser‐scanning microscopy showed that the dairy‐based gluten‐free bread crumb contained network‐like structures resembling the gluten network in wheat bread crumb. It was concluded that the formation of a continuous protein phase is critical for an improved keeping quality of gluten‐free bread.     

Fractionation and reconstitution experiments provide insight into the role of gluten and starch interactions in pasta quality

Commercial durum wheat (Triticum durum desf.) semolina was fractionated into starch, gluten, and water extractables. Starch surface proteins and surface lipids were removed, and two starches with manipulated granule size distributions were produced to influence starch properties, affecting its interaction with other semolina components. Reconstituted spaghetti was made with untreated (control) or treated starches. The pasta made from the starting semolina material had lower cooking time and was of lower quality than the samples made from reconstituted material. This was not due to changes in gluten properties as a result of the first step of the fractionation process. For the reconstituted samples, starch interaction behavior was not changed after surface protein or surface lipid removal. Starch surface properties thus do not influence the starch interaction behavior, indicating that starch-gluten interaction in raw (uncooked) pasta is mainly due to physical inclusion. All reconstituted pasta samples also had generally the same cooking quality. It was concluded that the small changes in starch gelatinization behavior, caused by the above-mentioned starch modifications, are of little importance for pasta quality.     

Grain Quality Characteristics and Milling Performance of Full and Partial Waxy Durum Lines

Mutation of the gene coding for the granule bound starch synthase (waxy protein) leads to reduced amylose content in cereal endosperm. Durum wheat (Triticum turgidum L. var. durum) has one waxy locus in each of its two genomes. Full waxy durum wheat is produced when both genomes carry the waxy null alleles. When only one locus is mutated, partial waxy durum wheat is obtained. Partial and full waxy near‐isogenic lines of durum wheat developed by a breeding program were analyzed as to their quality characteristics. Amylose was largely eliminated in full waxy lines; however, no reduction in amylose content was detected in partial waxy lines. The waxy mutation did not affect grain yield, kernel size, or kernel hardness. Full waxy durum lines had higher kernel ash content, α‐amylase activity, and a unique nonvitreous kernel appearance. Protein quality, as evaluated by SDS microsedimentation value, gluten index, and wet gluten was slightly lower in the full waxy lines than in the other genotypes. However, comparisons with current cultivars indicated that protein quality of all derived lines remained in the range of strong gluten cultivars. Semolina yield was lowered by the waxy mutations due to lower friability that resulted in less complete separation of the endosperm from the bran. Waxy semolina was more sensitive to mechanical damage during milling, but modified tempering and milling conditions may limit the damage. Overall, quality characteristics of waxy durum grain were satisfactory and suitable for application testing.     

Extractability and Molecular Modifications of Gliadin and Glutenin Proteins Withdrawn from Different Stages of a Commercial Ethanol Fuel/Distillers Dried Grains with Solubles Process Using a Wheat Feedstock

Extractability and molecular modifications of gliadin and glutenin proteins withdrawn from different stages of a commercial ethanol fuel/distillers dried grains with solubles (DDGS) process using a wheat feedstock were investigated. Materials were taken postliquefaction (PL), postdistillation (whole stillage), and postdrying (DDGS) during the process and then fractionated to separate the gliadins and the soluble high‐ and low‐molecular‐weight glutenins following a modified Verbruggen extraction method. Each fraction was characterized based on the extraction efficiencies within various aqueous alcohols of propan‐1‐ol, electrophoretic patterns, intrinsic and extrinsic fluorescence, free and total sulfhydryl content, and total disulfide bond levels. Findings indicated significant changes to the composition of extracted proteins and modifications to the protein structure (i.e., surface properties and conformation) throughout the ethanol/DDGS process, beginning with the first step of production (PL, ≈83°C). Overall, processing resulted in a shift toward an unextractable gluten matrix, accompanied by increases in hydrophobicity, disulfide bridging, and excessive protein aggregation.