Molecular characterization of storage proteins for selected durum wheat varieties grown in different environments

The variations of the amounts of individual high molecular weight glutenin subunits (HMW-GS), of the ratios HMW-GSy to HMW-GSx and HMW-GS to low molecular weight glutenin subunits (LMW-GS) and of protein content were evaluated for eight durum wheat cultivars in two regions using four fertilizer combinations during two successive years. All measured parameters showed significant variation with genotypes (G), environments (E) and fertilizers (F). The interaction E × G × F was highly significant for glutenin amount variation. Amongst cultivars possessing HMW-GS 20, landraces seem to better value the N-fertilizer use for the accumulation of HMW-GSy than high yielding cultivars. Both HMW-GSy to HMW-GSx and HMW-GS to LMW-GS ratios were found to be positively correlated (p < 0.05) with total protein content.     

Polymerisation of gluten proteins in developing wheat grain as affected by desiccation

The breadmaking quality of wheat is affected by the composition of gluten proteins and the polymerisation of subunits that are synthesised and accumulated in developing wheat grain. The biological mechanisms and time course of these events during grain development are documented, but not widely confirmed. Therefore, the aim of this study was to monitor the accumulation of gluten protein subunits and the size distribution of protein aggregates during grain development. The effect of desiccation on the polymerisation of gluten proteins and the functional properties of gluten were also studied. The results showed that the size of glutenin polymers remained consistently low until yellow ripeness (YR), while it increased during grain desiccation after YR. Hence, this polymerisation process was presumed to be initiated by desiccation. A similar polymerisation event was also observed when premature grains were dried artificially. The composition of gluten proteins, the ratios of glutenin to gliadin and high molecular weight-glutenin subunits to low molecular weight-glutenin subunits, in premature grain after artificial desiccation showed close association with the size of glutenin polymers in artificially dried grain. Functional properties of gluten in these samples were also associated with polymer size after artificial desiccation.     

Effect of high molecular weight glutenins and rye translocations on soft wheat flour cookie quality

The influence of high molecular weight glutenin subunits (HMW-GS) on wheat breadmaking quality has been extensively studied but the effect of different Glu-1 alleles on cookie quality is still poorly understood. This study was conducted to analyze the effect of HMW-GS composition and wheat-rye translocations on physicochemical flour properties and cookie quality of soft wheat flours. Alleles encoded at Glu-A1, Glu-B1 and Glu-D1 locus had a significant effect over physicochemical flour properties and solvent retention capacity (SRC) profile. The null allele for Glu-A1 locus presented the highest cookie factor observed (CF = 7.10), whereas 1BL/1RS and 1AL/1RS rye translocations had a negative influence on CF. The three cultivars that showed the highest CF (19, 44 and 47) had the following combination: Glu-A1 = null, Glu-B1 = 7 + 8, Glu-D1 = 2 + 12 and no secalins. Two prediction equations were developed to estimate soft wheat CF: one using the HMW-GS composition and the other using physicochemical flour parameters, where SRCsuc, SRC carb, water-soluble pentosans, damaged starch and protein turned out to be better CF predictors. This data suggests that grain protein allelic composition and physicochemical flour properties can be useful tools in breeding programs to select soft wheat of good cookie making quality.     

Study of the mechanism of improvement due to waxy wheat flour addition on the quality of frozen dough bread

Waxy wheat flour (WWF) was substituted for 10% regular wheat flour (RWF) in frozen doughs and the physicochemical properties of starch and protein isolated from the frozen doughs stored for different time intervals (0, 1, 2, 4 and 8 weeks) were determined to establish the underlying reasons leading to the effects observed in WWF addition on frozen dough quality. Using Nuclear Magnetic Resonance (NMR), Differential Scanning Calorimeter (DSC) and X-ray Diffraction (XRD) among others, the gluten content, water molecular state, glutenin macropolymer content, damaged starch content, starch swelling power, gelatinization properties, starch crystallinity and bread specific volume were measured. Compared to RWF dough at the same frozen storage condition, 10% WWF addition decreased dry gluten and glutenin macropolymer contents and T₂₃ proton density of frozen dough, but increased the wet gluten content, T₂₁ and T₂₂ proton density. 10% WWF addition also decreased damaged starch content, but increased starch swelling power, gelatinization temperature and enthalpy, crystallinity of starch and bread specific volume of frozen dough. Results in the present study showed that the improvement observed due to WWF addition in frozen dough bread quality might be attributed to its inhibition of redistribution of water molecules bound to proteins, increase in damaged starch content and decrease in starch swelling power.     

Hydration,polymerization and rheological properties of frozen gluten-water dough as influenced by thermostable ice structuring protein extract from Chinese privet (Ligustrum vulgare) leaves

The effects of thermostable ice structuring proteins (TSISPs) extracted from Chinese privet (Ligustrum vulgare) leaves on water molecular state, dehydration of gluten proteins, secondary structure of proteins, glutenin subunit of glutenin macropolymer (GMP) and rheological properties of gluten doughs during frozen storage were investigated by nuclear magnetic resonance (NMR), attenuated total reflectance-Fourier transform infrared reflectance (ATR-FTIR), reversed phase-high performance liquid chromatography (RP-HPLC) and dynamic rheometry. After frozen storage for 5 weeks, the control sample showed dehydration of gluten proteins and mobility of water molecules in gluten dough increased, significantly indicating ice formation and water redistribution. Secondary structure of gluten proteins changed significantly, α-helix decreased and β-sheet increased. Glutenin subunits depolymerized, indicated by the decrease in high molecular weight glutenins/low molecular weight-glutenins (HMW/LMW) ratio. The decrease in elastic moduli (G′) and viscous moduli (G′') showed the deterioration of rheological properties of gluten dough. The addition of TSISPs inhibited the dehydration of gluten proteins, decrease in α-helix, increase in β-sheet and HMW/LMW ratio, resulting in improved rheological properties of gluten dough.     

Effect of allelic variation at the Glu-3/Gli-1 loci on breadmaking quality parameters in hexaploid wheat (Triticum aestivum L.)

Low molecular weight glutenin subunits (LMW-GS) encoded by the Glu-3 loci are known to contribute to wheat breadmaking quality. However, the specific effect of individual Glu-3 alleles is not well understood due to their complex protein banding patterns in SDS-PAGE and tight linkage with gliadins at the Gli-1 locus. Using DNA markers and a backcross program, we developed a set of nine near isogenic lines (NILs) including different Glu-A3/Gli-A1 or Glu-B3/Gli-B1 alleles in the genetic background of the Argentine variety ProINTA Imperial. The nine NILs and the control were evaluated in three different field trials in Argentina. Significant genotype-by-environment interactions were detected for most quality parameters indicating that the effects of the Glu-3/Gli-1 alleles are modulated by environmental differences. None of the NILs showed differences in total flour protein content, but relative changes in the abundance of particular classes of proteins cannot be ruled out. On average, the Glu-A3f, Glu-B3b, Glu-B3g and Glu-B3i_Man alleles were associated with the highest values in gluten strength-related parameters, while Glu-A3e, Glu-B3a and Glu-B3i_Chu were consistently associated with weak gluten and low quality values. The value of different Glu-3/Gli-1 allele combinations to improve breadmaking quality is discussed.     

Disulphide structure of high-molecular-weight (HMW-) gliadins as affected by terminators

This study aimed at elucidating SS-bonds of HMW-gliadins (HGL) from wheat with the focus on terminators of glutenin polymerisation. HGL from wheat flour extracts non-treated or treated with the S-alkylation reagent N-ethylmaleinimide (NEMI) were compared. HGL from wheat flour Akteur were isolated, hydrolysed with thermolysin and the resulting peptides pre-separated by gel permeation chromatography and analysed by liquid chromatography/mass-spectrometry using alternating electron transfer dissociation/collision-induced dissociation. Altogether, 22 and 28 SS-peptides from samples without and with NEMI treatment, respectively, were identified. Twenty-six peptides included standard SS-bonds of α- and γ-gliadins, high-molecular-weight and low-molecular-weight glutenin subunits. Eleven SS-bonds were identified for the first time. Fifteen peptides unique to HGL contained cysteine residues from gliadins with an odd number of cysteines (ω5-, α- and γ-gliadins). Thus, gliadins with an odd number of cysteines, glutathione and cysteine had acted as terminators of glutenin polymerisation. Decisive differences between samples without and with NEMI treatment were not obvious showing that the termination of polymerisation was already completed in the flour. The two HGL samples, however, were different in the majority of ten peptides that included disulphide-linked low-molecular-weight (LMW) thiols such as glutathione and cysteine with the former being enriched in the non-treated HGL-sample.     

Immunomodulatory activities of non-prolamin proteins in wheat germ and gluten

Albumin (Alb), globulin (Glo), glutelin (Gll) and glutenin (Gln) were separately extracted from wheat germ and wheat gluten. Amino acisd composition, molecular weight distribution, solubility, in vitro digestibility, and immunomodulatory activities were all analyzed. Gll and Gln have similar molecular weight distributions, which differed from those of Alb and Glo. Alb showed the highest solubility at various pH values (except pH 4.0), whereas Glo showed the highest in vitro digestibility. Glo and Gll have the highest proportion of essential to total amino acids, while Alb and Gll have the highest protein digestibility-corrected amino acid scores. Gll had the strongest immunomodulatory effects in terms of stimulation of RAW 264.7 cells to produce IL-6, TNF-α, and IL-10, and good stimulatory effects on splenocyte proliferation, production of IL-2, phagocytosis, and secretion of nitric oxide in RAW 264.7 cells. Gll can be considered a good protein source for use in health foods.     

The impact of redox agents on further dough development,relaxation and elastic recoil during lamination and fermentation of multi-layered pastry dough

The role of gluten proteins during lamination and fermentation of multi-layered wheat flour pastry dough was examined by including oxidizing or reducing agents in the recipe to respectively strengthen or weaken the gluten protein network. Pastry burst rig textural measurements showed that dough strength increases during lamination up to 16 fat layers. However, further lamination up to 64 and 128 fat layers decreases the dough strength, most likely due to destruction of layer integrity. Redox agents strongly affect dough strength. Furthermore, fermentation and spread tests showed that they strongly influence elastic recoil immediately after lamination and during relaxation. Moreover, elastic recoil consistently occurs to a greater extent in the final direction of sheeting. None of the observed changes in dough strength and relaxation behaviour could be linked to changes in the levels of protein extractable in sodium dodecyl sulfate containing medium (SDS-EP). This suggests that changes occur preferentially either within the SDS-extractable or within the non-SDS-EP fraction and that they do not render non-extractable protein fractions extractable or vice versa. Furthermore, elastic recoil is most likely caused by reformation of inter- and intramolecular hydrogen bonds and hydrophobic interactions.     

Impact of lipases with different substrate specificity in wheat flour separation on the properties of the resultant gluten

Wheat gluten was isolated in a laboratory dough-batter flour separation process in the presence or absence of lipases differing in hydrolysis specificity. The obtained gluten was blended with wheat starch to obtain gluten-starch (GS) blends of which the water and oil binding capacities were investigated. Furthermore, GS blends were mixed into dough and processed into model breads, of which dough extensibility and loaf volume were measured, respectively. In comparison to GS blends prepared with control gluten, oil binding capacity was higher when GS blends contained gluten isolated with Lecitase Ultra (at 5.0 mg enzyme protein/kg flour), a lipase hydrolyzing both non-polar and polar lipids. Additionally, dough extensibility and total work needed for fracture were lower for dough prepared from GS blends containing gluten isolated with Lipolase (at 5.0 mg enzyme protein/kg flour), a lipase selectively degrading non-polar lipids. In GS blend bread making, this resulted in inferior loaf volumes. Comparable GS blend properties were measured when using control gluten and gluten isolated with YieldMAX, a lipase mainly degrading N-acyl phosphatidylethanolamine. In conclusion, properties of GS blend model systems are altered when gluten prepared in the presence of lipases is used to a degree which depends on lipase specificity and concentration.     

Extractability and chromatographic separation of rye (Secale cereale L.) flour proteins

A size exclusion – high performance liquid chromatography (SE-HPLC) method originally developed for separating wheat, barley or rice proteins was applied to study the extractability and molecular weight (MW) distribution of rye flour proteins. These were extracted with 50 mmol/l sodium phosphate buffer (pH 6.8) containing 2.0% (w/v) sodium dodecyl sulfate (SDS) and, optionally, 1.0% (w/v) dithiothreitol (DTT). About 95% of the proteins were extracted in buffer containing 2.0% SDS. Addition of 1.0% DTT to such buffer increased the protein extractability to 100%, indicating that rye flour contains some proteins cross-linked by disulfide (SS) bonds. The SE-HPLC profiles revealed that rye flour contains SS-linked HMW-secalins and 75 k γ-secalins which elute in specific peaks. Upon reduction, these SS-linked protein aggregates dissociate and some entrapped albumins, globulins and/or ω-secalins are released. Rye flour albumins and globulins elute over the entire SE-HPLC profile. In contrast, the monomeric ω-secalins and 40 k γ-secalins are detected in specific well resolved SE-HPLC peaks. The applied fast and reproducible method can be used to characterise and quantify rye flour proteins and to determine changes as a result of processing.     

Modification of physicochemical properties and in vitro digestibility of wheat flour through superheated steam processing

Native and moistened wheat flours (moisture contents were 13.5 and 27.0%, respectively) were treated with superheated steam (SS) at different temperatures (140 and 170 °C) and times (1, 2 and 4 min). Their physicochemical and digestive properties were analyzed. For native flour, SS treatment altered the starch molecular structure and behavior slightly. While for moistened flour, crystalline degree, gelatinization enthalpy, amylose leaching (AML) and falling number significantly decreased, but thermal transition temperatures increased with the rise of treating severity. Clumping of starch granules, aggregation of proteins and formation of amylose-lipid complexes occurred in both native and moistened flours. Broader pasting temperature ranges and higher viscosities were found on SS-modified flours. Additionally, SS treatment on moistened flours increased resistant and slowly digestible starch contents. In general, SS treatment induced changes in starch molecular structure and reactions among flour components leading to more stable structures, thus affecting their pasting behavior, thermal properties and in vitro digestion.     

Kernel vitreousness and protein content: Relationship,interaction and synergistic effects on durum wheat quality

Four sets of durum samples were used in this study to further understand the interrelationships among hard vitreous kernels (HVK), protein content, and pigment concentration, with a focus on the interaction and synergistic effects of protein content and vitreousness on durum quality. HVK level increases with higher protein content in the range of 9.5–12.5%, but this relationship is less evident in durum samples with high protein content (12.5–14.5%). Both protein content and kernel vitreousness can significantly affect durum milling quality. White starchy kernels (WSK) in low protein durum have a very detrimental impact on milling and pasta processing quality, but high protein content can mitigate the adverse impact of WSK on durum quality. Although protein content plays a dominant role, higher HVK might contribute positively to pasta firmness. There was no significant difference in yellow pigment content between HVK and WSK. However, pigment loss from semolina to dough was higher for WSK than HVK. Despite the difference in protein content, HVK and WSK have little difference in gluten strength. The monomeric protein was preferentially accumulated in HVK. The glutenin proteins of HVK and WSK were similar in the ratios of 1Bx/1By and HMW/LMW-GS.     

Influence of allelic prolamin variation and localities on durum wheat quality

Thirty-seven varieties of a Mediterranean durum wheat collection grown in Tunisia and Spain were analysed for their allelic composition in prolamins, as well as their protein concentration, sodium dodecyl sulphate sedimentation (SDSS) test and mixograph parameters. Genotype was a greater source of variation in all measurements than locality. Uncommon high and low molecular glutenin subunits (HMW-GS and LMW-GS) were found (V and 2•• subunits at Glu-A1, 13 + 16 at Glu-B1, 5* subunit and ax allele at Glu-A3). The rare combinations 2 + 4+14 + 18 and 8 + 9+13 + 16+18 subunits at the Glu-B3 locus were found. Glu-A3ax had a positive influence on SDSS and mixograph parameters. Of all the prolamins, those that have the B-LMW-GS composition aaa (for Glu-A3, Glu-B3 and Glu-B2 loci, respectively), when associated with the Glu-A1c and Glu-B1d gave the best semolina quality. By contrast, semolina quality is poor when this same composition is associated with the Glu-A1c and Glu-B1e and even poorer when associated with the Glu-A1c and Glu-B1f. In addition, the cultivars with B-LMW-GS allelic composition aab (for Glu-A3, Glu-B3 and Glu-B2 loci, respectively), when associated with the Glu-A1c and Glu-B1d, gave high quality, whereas when associated with the Glu-A1c and Glu-B1e or with Glu-A1o and Glu-B1f, the quality was very poor.     

Dough rheological properties and noodle-making performance of non-waxy and waxy whole-wheat flour blends

Dough rheological properties and noodle-making performance of non-waxy whole-wheat flour (WWF) with partial- or full-waxy (PW- or FW-) WWF substitution were studied. The substitution levels were 0, 250, 500, 750, and 1000 g/kg, respectively. FW-WWF reduced the peak viscosity and pasting temperature of WWF blends as its substitution level was increased due to its higher proportions of B-type starch granules and short amylopectin chains, while PW-WWF increased peak viscosity with the increasing substitution level because of its higher amylopectin content. As demonstrated by farinograph and rheometer measurements, FW-WWF interfered with gluten development because of the increased competition for water by arabinoxylans and amylopectin; however, PW-WWF enhanced dough strength due primarily to its increased protein content. Consequently, FW-WWF showed a detrimental effect on cooked noodle texture as the cooked noodle hardness was reduced by 50% at the 1000 g/kg substitution level. In contrast, PW-WWF enhanced noodle integrity and elasticity by increasing cooked noodle cohesiveness and resilience by 10.1% and 14.8%, respectively, at the 1000 g/kg substitution level. The results suggest that with waxy WWF substitution, the changes in starch composition, arabinoxylans, and protein content could modify the interactions among flour components and influence the quality characteristics of noodle products.     

Effect of physicochemical and empirical rheological wheat flour properties on quality parameters of bread made from pre-fermented frozen dough

The objective of this study was to examine the influence of flour quality on the properties of bread made from pre-fermented frozen dough. The physicochemical parameters of 8 different wheat flours were determined, especially the protein quality was analysed in detail by a RP-HPLC procedure. A standardized baking experiment was performed with frozen storage periods from 1 to 168 days. Baked bread was characterised for specific loaf volume, crumb firmness and crumb elasticity. The results were compared to none frozen control breads. Duration of frozen storage significantly affected specific loaf volume and crumb firmness. The reduction of specific loaf volume was different among the used flours and its behaviour and intensity was highly influenced by flour properties. For control breads wet gluten, flourgraph E7 maximum resistance and RVA peak viscosity were positively correlated with specific loaf volume. However, after 1–28 days of frozen storage, wet gluten content was not significantly influencing specific loaf volume, while other parameters were still significantly correlated with the final bread properties. After 168 days of frozen storage all breads showed low volume and high crumb firmness, thus no significant correlations between flour properties and bread quality were found. Findings suggest that flours with strong gluten networks, which show high resistance to extension, are most suitable for frozen dough production. Furthermore, starch pasting characteristics were also affecting bread quality in pre-fermented frozen dough.     

Variation in gluten quality parameters of spring wheat varieties of different origin grown in contrasting environments

The aim of this study was to investigate variation in protein content and gluten viscoelastic properties in wheat genotypes grown in two mega-environments of contrasting climates: the southeast of Norway and Minnesota, USA. Twelve spring wheat varieties, nine from Norway and three HRS from Minnesota, were grown in field experiments at different locations in Norway and Minnesota during 2009–2011. The results showed higher protein content but lower TW and TKW when plants were grown in Minnesota, while the gluten quality measured as Rmax showed large variation between locations in both mega-environments. On average, Rmax of the samples grown in Minnesota was higher than those grown in Norway, but some locations in Norway had similar Rmax values to locations in Minnesota. The data showed inconsistent relationship between the temperature during grain filling and Rmax. Our results suggest that the weakening effect of low temperatures on gluten reported in this study are caused by other environmental factors that relate to low temperatures. The variety Berserk showed higher stability in Rmax as it obtained higher values in the environments in Norway that gave very weak gluten for other varieties.     

Combined mutations in five wheat STARCH BRANCHING ENZYME II genes improve resistant starch but affect grain yield and bread-making quality

Increases in the proportion of amylose in the starch of wheat grains result in higher levels of resistant starch, a fermentable dietary fiber associated with human health benefits. The objective of this study was to assess the effect of combined mutations in five STARCH BRANCHING ENZYME II (SBEII) genes on starch composition, grain yield and bread-making quality in two hexaploid wheat varieties. Significantly higher amylose (∼60%) and resistant starch content (10-fold) was detected in the SBEII mutants than in the wild-type controls. Mutant lines showed a significant decrease in total starch (6%), kernel weight (3%) and total grain yield (6%). Effects of the mutations in bread-making quality included increases in grain hardness, starch damage, water absorption and flour protein content; and reductions in flour extraction, farinograph development and stability times, starch viscosity, and loaf volume. Several traits showed significant interactions between genotypes, varieties, and environments, suggesting that some of the negative impacts of the combined SBEII mutations can be ameliorated by adequate selection of genetic background and growing location. The deployment of wheat varieties with increased resistant starch will likely require economic incentives to compensate growers and millers for the significant reductions detected in grain and flour yields.     

The impact of Septoria tritici blotch in bread making quality among argentinean wheat cultivars

Septoria tritici blotch (STB), caused by Zymoseptoria tritici is a relevant foliar wheat disease worldwide. Several reports show the importance of STB on grain yield, their components and grain protein while little is known about its effect on the rheological properties of the wheat flour. The scarce literature found, only mentions the effect of the complex of foliar diseases on wheat quality, without individualizing the effect of the different pathogens separately. This study analyze the influence of increasing doses of inoculum of Zymoseptoria tritici, on the bread making quality of ten Argentinean wheat cultivars and its possible variation according to their quality group. The increase of inoculum concentration augmented the area under disease progress curve, decreased green flag leaf area duration and green leaf area duration. Cultivars K. Flecha and B.75 Aniversario had the lowest green flag leaf area duration causing higher reduction in grain filling period and higher reductions in P, indicating a lower gliadin/glutenin ratio. STB decreased P/L and E while L, W, D, SV and bread volume increased. Cultivars differed in rheological parameters according to their quality group. Gluten/protein relationship was significant in quality group 1 and non-significant in cultivars belonging to quality group 2 and 3.     

Determination of volatile compounds in heat-treated straight-grade flours from normal and waxy wheats

Volatile compounds formed during heat-treatment of wheat flour influence the application of treated flour. In this study, normal and waxy hard wheat flours before and after dry-heat treatment were subjected to headspace analysis by solid-phase microextraction of volatiles followed gas chromatography–mass spectrometry (GC/MS). The untreated waxy wheat flour contained higher levels of odor-active compounds than normal wheat flour including aldehydes, alcohols, furans, and ketones. Lipid oxidation appears to play major role in producing such odor compounds. Heat treatments, depending on the severity, alter the profile of volatile compounds. Low temperature (100–110 °C) treatments effectively eliminated cereal odor (aldehyde) and did not introduce additional odors, providing a possible way to produce low-odor flours. Heat treatments at 120 °C and higher temperatures elevated the content of pyrazines, furans, and sulfur-containing compounds which together gave a roasty aroma to the flours. Considering organoleptic properties, treatments of flours at 140 °C was superior to 160 °C. The waxy wheat flour was more prone to produce odor-active compounds than normal wheat flour during the same heat treatment.