Relationship Between Rheological Properties and Microstructural Characteristics of Nondeveloped,Partially Developed,and Developed Doughs

Farinography and mixography are two commonly used procedures for evaluating dough properties. These procedures, however, cannot separate hydration and energy input during dough development, both of which are critically important for understanding fundamental rheological properties of dough. A rheometer and laser scanning confocal microscopy (LSCM) were used to study the relationship between rheological properties and microstructural characteristics of developed (by farinograph with both shear and extensional deformations), of partially developed (by rheometer with either shear or extensional deformation), and of nondeveloped (no deformation) dough samples of wheat flours. Rheological data revealed that developed dough had the highest G* (most elastic or strong), followed by doughs partially developed with extensional deformation, and then shear deformation, and finally by nondeveloped dough. The LSCM z‐sectioning (scanning of different layers of the sample) and the analysis of amount of protein matrix showed that developed dough had the most protein matrix and nondeveloped dough had the least protein matrix. It also showed that the higher the G*, the greater the protein network. Moreover, the type of deformation appeared to contribute to the development of protein matrix and further increase the dough strength. In this study, a combination of shear and extensional deformations by farinograph produced the most protein matrix and the strongest dough, followed by extensional deformation, shear deformation, and then no deformation.     

Quantitative Glutenin Composition from Gel Electrophoresis of Flour Mill Streams and Relationship to Breadmaking Quality

Three samples of Nekota (hard red winter wheat) were milled, and six mill streams were collected from each sample. The 18 mill streams were analyzed separately as well as recombined to form three patent flours. The methods of multistacking (MS)‐SDS‐PAGE and SDS‐PAGE were used to separate the unreduced SDS‐soluble glutenins and the total reduced proteins, respectively. The separated proteins were quantified by densitometry. The quantity of unreduced SDS‐soluble proteins was significantly different among the mill streams at the 4% (largest molecular weight polymeric glutenins) and at the 10 and 12% (smaller molecular weight polymeric glutenins) origins of the MS‐SDS‐PAGE gels. The quantities of total HMW‐GS, LMW‐GS, 2*, 7+9, and 5+10 subunits and the ratio of HMW‐GS to LMW‐GS in polymeric protein samples isolated using preparative MS‐SDS‐PAGE and in total reduced protein extracts were significantly different among mill streams. The quantities of HMW‐GS, LMW‐GS, 2*, 7+9, and 5+10 subunits from total reduced proteins were positively and significantly correlated with loaf volume. The quantities of glutenin subunits (both HMW‐GS and LMW‐GS) from unreduced SDS‐soluble proteins were positively or negatively correlated with loaf volume at the various MS‐SDS‐PAGE gel origins but the levels of correlation were not significant. These results showed that the glutenin protein composition was different among the various mill streams and demonstrated that electrophoretic analysis of the proteins in these fractions is a useful tool for studying the variation in functional properties of flour mill streams.     

Quantitative Variation of HMW Glutenin Subunits from Hard Red Spring Wheats Grown in Different Environments

The objective of this study was to investigate the quantitative variation of HMW glutenin subunits in relation to glutenin polymers and hence breadmaking quality across different environments. Six genotypes of hard red spring (HRS) wheat were grown at seven locations in North Dakota in 1998 in a randomized complete‐block experimental design with three replicates at each location. Unreduced SDS‐soluble glutenins of flour were fractionated by multistacking SDS‐PAGE into different sized glutenin polymers, followed by SDS‐PAGE and imaging densitometry to determine the quantitative variation of HMW glutenin subunits. SDS‐insoluble glutenin polymers also were examined for their quantitative composition of HMW glutenin subunits. The results showed that the percentage of HMW glutenin subunits was significantly affected by growing locations. The quantity of HMW glutenin subunits in SDS‐insoluble glutenins was significantly and positively correlated with loaf volume. SDS‐insoluble glutenin polymers had a higher percentage of HMW glutenin subunits than did SDS‐soluble glutenins. SDS‐insoluble glutenin polymers in flour were positively and significantly correlated in proportions of both total and individual HMW glutenin subunits in total SDS glutenins. SDS‐insoluble glutenin polymers also were positively and significantly correlated with the combined proportion of HMW glutenin subunits 2* + 5. The results of this study indicated that either subunit 2* or 5 might be more important in forming a greater quantity of larger SDS‐insoluble glutenin polymers than other subunits. SDS‐insoluble glutenin polymers from different cultivars or locations could have different quantities of HMW glutenin subunits in their composition. SDS‐insoluble glutenin polymers with more HMW glutenin subunits might be larger sized than those with less HMW glutenin subunits. Environment significantly influenced the quantitative variation of HMW glutenin subunits, which in turn affected the size distribution of glutenin polymers, and hence breadmaking quality.     

Studies on Effects of Microbial Transglutaminase on Gluten Proteins of Wheat. I. Biochemical Analysis

The enzyme transglutaminase (TG) is known to have beneficial effects on breadmaking. However, only limited information is available on the structural changes of gluten proteins caused by TG treatment. The effect of TG has, therefore, been systematically studied by means of model peptides, suspensions of wheat flours and doughs. The treatment of synthetic peptides mimicking amino acid sequences of HMW subunits of glutenin with TG results in isopeptide bonds between glutamine and lysine residues. To study the effect on gluten proteins, different amounts of TG (0 to 900 mg enzyme protein per kg) were dissolved in a buffer and added to wheat flour. The flour suspensions were incubated and centrifuged and the residues were successively extracted with water, a salt solution, 60% aqueous ethanol (gliadin fraction) and SDS solution including a reducing agent (glutenin fraction). The characterization of the fractions by amino acid analysis, SDS‐PAGE, gel permeation HPLC and reversed‐phase HPLC has indicated that the quantity of extractable gliadins decreases by increasing TG amounts. Among gliadins, the ω5‐type was affected to the greatest extent by the reduction of extractability, followed by the ω1,2‐, α‐ and γ‐types. The oligomeric portion of the gliadin fractions (HMW gliadin) was strongly reduced when flour was treated with 450 and 900 mg TG per kg of flour, respectively. In the first instance, the quantity of the glutenin fractions increased by the treatment of flour with 90 and 450 mg TG per kg of flour, and significantly decreased by the treatment of flour with 900 mg TG per kg of flour. Parallel to an increase in TG concentration, the amounts of glutenin‐bound ω‐gliadins and HMW subunits were strongly reduced, whereas the LMW subunits reached a maximal amount after treatment with 450 mg TG per kg of flour. The insoluble residue was almost free of protein when flour was treated with lower amounts of TG. Higher amounts led to a great increase of protein in the residues. The effects of TG on doughs were similar to those of flour suspensions, but less strongly pronounced probably due to the lower water content of the dough system. Sequence analysis of peptides from a thermolytic digest of the insoluble residue revealed that HMW subunits of glutenin and α‐gliadins were predominantly involved in cross‐links formed by TG treatment.     

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.     

Effects of Genotype and Environment on Glutenin Polymers and Breadmaking Quality

Six genotypes of hard red spring (HRS) wheat were grown at seven environments in North Dakota during 1998. Effects of genotype and environment on glutenin polymeric proteins and dough mixing and baking properties were examined. Genotype, environment, and genotype‐by‐environment interaction all significantly affected protein and dough mixing properties. However, different protein and quality measurements showed differences for relative influences of genotype and environment. Total flour protein content and SDS‐soluble glutenin content were influenced more by environmental than genetic factors, while SDS‐insoluble glutenin content was controlled more by genetic than environmental factors. Significant genotypic and environmental effects were found for the size distribution of SDS‐soluble glutenins and between SDS‐soluble and SDS‐insoluble glutenins as well as % SDS‐insoluble glutenins. With increased flour protein content, the proportions of monomeric proteins and SDS‐insoluble glutenin polymers appeared to increase, but SDS‐soluble glutenins decreased. Flour protein content and the size distribution between SDS‐soluble and SDS‐insoluble glutenin polymers were significantly correlated with dough mixing properties. Environment affected not only total flour protein content but also the content of different protein fractions and size distributions of glutenin polymers, which, in turn, influenced properties of dough mixing. Flour protein content, % SDS‐insoluble glutenin polymers in flour, and ratio of SDS‐soluble to SDS‐insoluble glutenins all were highly associated with dough mixing properties and loaf volume.     

Relationship Between Physicochemical Properties of Wheat Flour,Wheat Protein Composition,and Textural Properties of Cooked Chinese White Salted Noodles

Physicochemical properties and protein composition of 39 selected wheat flour samples were evaluated and correlated with the textural properties of Chinese hard‐bite white salted noodles. Flour samples were analyzed for their protein and wet gluten contents, sedimentation volume, starch pasting properties, and dough mixing properties by farinograph and extensigraph. Molecular weight distribution of wheat flour proteins was determined with size‐exclusion (SE) HPLC, SDS‐PAGE, and acid‐PAGE. Textural properties of Chinese hard‐bite white salted noodles were determined through texture profile analysis (TPA). Hardness, springiness, gumminess, and chewiness of cooked noodles were found to be related to the dough mixing properties. Both protein content and protein composition were found to be related to TPA parameters of noodles. The amount of total flour protein was positively correlated to hardness, gumminess, and chewiness of noodles. The absolute amounts of different peak proteins obtained from SE‐HPLC data showed positive correlations with the hardness, gumminess, chewiness, and springiness of noodles. The proportions of these peak proteins were, however, not significantly related to texture parameters. The proportions of low‐molecular‐weight glutenins/gliadins and albumins/globulins, as observed from SDS‐PAGE, were correlated positively and negatively, respectively, to the hardness, gumminess, and chewiness of cooked noodles. Among the alcohol‐soluble proteins (from acid‐PAGE data), β‐gliadins showed strong correlations with the texture properties of cooked noodles. For the selected flour samples, the total protein content of flour had a stronger relationship with the noodle texture properties than did the relative proportion of different protein subgroups. Prediction equations were developed for TPA parameters of cooked noodles with SE‐HPLC and rapid visco analysis data of the 30 flour samples, and it was found that about 75% of the variability in noodle hardness, gumminess, and chewiness values could be explained by protein composition and flour pasting properties combined together. About 50% of the variations in cohesiveness and springiness were accounted for by these prediction equations.     

Characterization and Quantification of Native Glutenin Aggregates by Multistacking Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) Procedures

Native glutenin aggregates of two different quality flours containing the same high molecular weight (HMW) glutenin subunit compositions were investigated by multistacking sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) procedures. Five stacking gels (4, 6, 8, 10, and 12%) with a separating (resolving) gel of 14% were used to separate nonreduced glutenin aggregates solubilized from flour by SDS sodium phosphate buffer. There were large differences in protein solubility of the two flours. It took 8 hr to extract 91% of total proteins from the strong flour (variety Len) while it took only 2 hr for the weak flour sample 205. Total glutenin proteins and the proportions of glutenins at the different origins (including the origin of the 14% separating gel) were quantified by high-resolution densitometry procedures. As the duration of extraction increased, both total glutenins and glutenins at the 4% origins increased. The good quality flour Len had a higher total glutenin protein (3% more) and higher proportion of glutenins with the largest molecular sizes (also 3% more) at the 4% origins than the poor quality flour sample 205. After glutenin aggregates from each origin were reduced and analyzed by SDS-PAGE, the largest glutenins at the 4% origin contained twice the amount of total HMW glutenin subunits when compared to the smaller aggregates at the 12% origin. Among the total HMW glutenin subunits, the proportion of subunit 5 (which published literature reports to be the largest molecular weight based on calculations of DNA-derived amino acid sequence analysis) was twice that at the 12% origin. A randomized structure of native glutenins is proposed based on the results of our investigations.     

Physicochemical and Rheological Characterization of Wheat Flour Dough

Rheological and structural behavior of dough prepared with two Argentinean flours (FI and FII) of different dough extensibilities were studied. Flours were analyzed by composition and rheological assays. Structural properties of dough prepared at different mixing times were analyzed by scanning electron microscopy, free sulfhydryls quantification, and yield of different protein fractions, as well as their protein surface hydrophobicity. Size of high molecular weight glutelin soluble aggregates was analyzed through multistacking gel electrophoresis. Dynamic viscoelasticity of dough was also studied. Flours FI and FII presented similar physicochemical properties but different rheological properties. Structural properties of both flour components were different. Starch from FI flour generated a more viscous paste than that of FII. FI presented a higher glutenin‐to‐gliadin ratio and a higher content of free sulfhydryls than FII. The resulting dough of FI showed a high development time and was more stable than FII. FI contained a high proportion of soluble HMW glutenins and formed dough with a more depolymerized insoluble protein residue containing a lower amount of gliadin in its matrix than FII. FI also formed a more elastic and stable dough with higher development time than FII. The specific structural characteristic of FI turns this flour into suitable raw material for the preparation of different bakery products in which elasticity of dough would be an important functional property.     

Effect of Hydrophilic Gums on the Quality of Frozen Dough: Electron Microscopy,Protein Solubility,and Electrophoresis Studies

Hydrophilic gums have been shown to improve the shelf‐life stability of frozen doughs during long periods of frozen storage. The objective of this research was to determine the effect of gums on starch and protein characteristics of frozen doughs using electron microscopy and electrophoresis studies. Frozen doughs, supplemented with three levels of gum arabic, carboxy methyl cellulose (CMC), kappa (κ) carrageenan, and locust bean gum, were studied after day 1 and after 4, 8, 12, and 16 weeks of frozen storage. Changes in the ultra structure of the frozen doughs were investigated, as well as the solubilities and composition of dough proteins by SDS‐PAGE. Scanning electron micrographs of doughs evaluated on day 0 (unfrozen) showed starch granules securely embedded in the gluten matrix. However, after 8 and 16 weeks of frozen storage, the frozen control dough without the gum additives clearly showed damage to the gluten network, and the starch granules appeared to be separated from the gluten. Doughs with locust bean gum and gum arabic showed better retention of the gluten network compared with the frozen control evaluated after different periods of storage. The SDS‐soluble protein content increased while residue protein content decreased as the frozen storage time increased. After each frozen storage period, the control dough without the gum additive had the highest amount of SDS‐soluble proteins and the lowest amount of residue proteins when compared with the doughs treated with gums. κ‐Carrageenan and locust bean gum had the lowest amount of SDS‐soluble proteins compared with doughs with CMC and gum arabic. The frozen control had the lowest amount of residue proteins at any particular time of frozen storage. κ‐Carrageenan treated doughs had the highest amount of residue proteins, followed by doughs with locust bean gum. Doughs with gum arabic and CMC had the lowest amount of residue proteins but still higher than the control doughs.     

Structural Modifications of Gluten Proteins in Strong and Weak Wheat Dough During Mixing

The network‐forming attributes of gluten have been investigated for decades, but no study has comprehensively addressed the differences in gluten network evolution between strong and weak wheat types (hard and soft wheat). This study monitored changes in SDS protein extractability, SDS‐accessible thiols, protein surface hydrophobicity, molecular weight distribution, and secondary structural features of proteins during mixing to bring out the molecular determinants of protein network formation in hard and soft wheat dough. Soft wheat flour and dough exhibited greater protein extractability and more accessible thiols than hard wheat flour and dough. The addition of the thiol‐blocking agent N‐ethylmaleimide (NEM) resulted in similar results for protein extractability and accessible thiols in hard and soft wheat samples. Soft wheat dough had greater protein surface hydrophobicity than hard wheat and exhibited a larger decrease in surface hydrophobicity in the presence of NEM. Formation of high‐molecular‐weight (HMW) protein in soft wheat dough was primarily because of formation of disulfides among low‐molecular‐weight (LMW) proteins, as indicated by the absence of changes in protein distribution when NEM was present, whereas in hard wheat dough the LMW fraction formed disulfide interaction with the HMW fraction. Fourier transform infrared spectroscopy indicated formation of β‐sheets in dough from either wheat type at peak mixing torque. Formation of β‐sheets in soft wheat dough appears to be driven by hydrophobic interactions, whereas disulfide linkages stabilize secondary structure elements in hard wheat dough.     

Isolation and Characterization of High‐Molecular‐Weight (HMW) Gliadins from Wheat Flour

The aim of this study was to isolate high‐molecular‐weight (HMW) gliadins from wheat flour and to characterize the protein components that contribute to HMW gliadins. Wheat flour Akteur was extracted with a modified Osborne procedure, and the fraction soluble in 60% ethanol (total gliadins) was separated by gel‐permeation HPLC, yielding three fractions, GP1–GP3. GP1 (21.5%) consisted of oligomeric HMW gliadins, GP2 (15.2%) of ω5‐gliadins, and GP3 (63.3%) of ω1,2‐, α‐, and γ‐gliadins. Two‐dimensional SDS‐PAGE of HMW gliadins showed that interchain disulfide bonds were present in HMW gliadins. The molecular mass distribution of HMW gliadins determined by gel‐permeation HPLC was in a range from 66,000 to 680,000 with an average degree of polymerization of 13. Reduced HMW gliadins were further separated by preparative reversed‐phase HPLC into four subfractions (RP1, RP2, RP3, and RP4), which were characterized by SDS‐PAGE and semiquantitative N‐terminal sequencing. HMW gliadins of the wheat flour Akteur contained all types of gluten proteins: 48% low‐molecular‐weight glutenin subunits, 18% γ‐gliadins, 13% α‐gliadins, 9% ω1,2‐gliadins, 8% HMW glutenin subunits, and 4% ω5‐gliadins. We postulate that the existence of HMW gliadins can be explained by the presence of terminators, which interrupt the polymerization of glutenin subunits during biosynthesis and lead to polymers of limited size (oligomers) that are still soluble in aqueous ethanol.     

Controlled Stepwise Reduction of Disulfide Bonds and Heat-Induced Modification of Wheat Dough Proteins

A reducing solution of 2-mercaptoethanol and its oxidized form 2-hydroxyethyl disulfide, whose variable concentrations set variable disulfide reduction potentials, was applied to progressively reduce the disulfide bonds of proteins extracted from doughs made from Meneba and Robin Hood flour. Several dough proteins had disulfide bonds stronger than those of other dough proteins. A SDS-sedimentation method was applied to monitor the baking of dough into bread. Dough proteins susceptible to heat (baking) were studied by SDS-fractionation, extraction with reducing alcoholic solution, SDS-PAGE, and N-terminal protein sequencing. High or low molecular weight glutenins, α, β, and γ-gliadins, α-amylase inhibitor, and α-amylase trypsin inhibitor were identified among the dough proteins modified by heat (as shown by reduced solubility in aqueous-SDS solution). The heat-induced modification of the gliadins and glutenins might contribute to the coagulation of dough proteins, while the heat-induced modification of the amylase or trypsin inhibitors might contribute to the regulation of endogenous or exogenous amylolytic or proteolytic activities in dough or bread.     

Lipid Extraction Process on Texturized Soy Flour and Wheat Gluten Protein‐Protein Interactions in a Dough Matrix

Protein‐protein interactions between wheat flour and solvent‐extracted (SE) or nonsolvent extracted (NSE) texturized soy flours were compared. Doughs were prepared to contain varying ratios of texturized soy flour in combination with wheat flour. Sucrose esters (2.5%) were included in several formulations. Doughs were fractionated into soluble and insoluble fractions at pH 4.7 and pH 6.1. Fractions were dried, powdered, and analyzed using SDS‐PAGE and spectrophotometric techniques. Electrophoretic evaluation indicated interactions between wheat gluten proteins and texturized soy proteins in the absence of sucrose esters. Electrophoretic gels of the wheat‐soy flour mixtures maintained a characteristic soy protein band after acidification to the soy protein isoelectric point. Inclusion of sucrose esters increased the interaction. Texturization conferred effects similar to that of sucrose ester on both forms of lipid‐extracted soy. Sulfhydryl analyses using 7‐chloro‐4‐nitrobenzo‐2‐oxa‐4, 3‐diazole (NBD‐Cl) revealed no change in the relative amount of sulfhydryl groups present in doughs prepared from either the texturized soy flours or the doughs containing equal amounts of wheat starch. These data indicate that interactions between soy protein from texturized soy flours and wheat proteins are not covalent.     

Genetic Variance for Gluten Strength Contributed by High Molecular Weight Glutenin Proteins

A total of 162 doubled haploid (DH) lines were produced from a cross between Triticum aestivum L. ‘AC Karma’ and line 87E03‐S2B1 to study the genetic contribution of high molecular weight (HMW) glutenin subunits to gluten strength. HMW glutenin subunit composition of each DH line was determined by SDS‐PAGE. The population was grown in the field at one location in 1999 and at three locations in 2000. Gluten strength and dough mixing properties were measured by mixograph test and SDS‐sedimentation test. Variance components were estimated for each measurement to determine the variability contributed by HMW glutenin subunits. Results indicated significant environmental impact on tested mixograph parameters, SDS‐sedimentation volumes and grain and flour protein concentration. Significant main effects of Glu‐1D loci encoded subunits were obtained for mixograph development time, energy to peak, slope after peak, and first minute slope. Lines containing 5+10 combination of subunits had higher values for mixograph development time and energy to peak, while slope after peak and first minute slope were lower as compared with 2+12 containing lines. Low intergenomic interactions were observed for bandwidth energy (BWE), total energy (TEG), and SDS‐sedimentation test, involving B and D genomes only. A portion of the genetic variability for gluten strength was accounted for overexpression of Bx7 subunit originating from the cultivar Glenlea derived line 87E03‐S2B1. There was no significant effect of Glu‐A1 encoded subunits on any of the tested parameters. Estimated genetic variability for gluten strength contributed by Glu‐B1 and Glu‐D1 encoded HMW glutenins was 55% for mixing development time and 51% for energy to peak.     

Effect of the Molecular Weight Distribution of Glutenin Protein from an Extra‐Strong Wheat Flour on Rheological and Breadmaking Properties Through Reconstitution Studies

Ten glutenin fractions were separated by sequential extraction of wheat gluten protein with dilute hydrochloric acid from defatted glutenin‐rich wheat gluten of the Canadian hard red spring wheat (HRSW) cultivar Glenlea. The molecular weight distribution (MWD) of 10 different soluble glutenin fractions was examined by multistacking SDS‐PAGE under nonreduced conditions. Also, the subunit composition of the different glutenin fractions was determined by SDS‐PAGE under reduced conditions. The MWD of the fractions (especially HMW glutenins) varied from fraction to fraction. From early to later fractions, the MWD shifted from low to high. The early extracted fractions contained more LMW glutenin subunits (LMW‐GS) and less HMW glutenin subunits (HMW‐GS). The later extracted fractions and the residue fraction contained much more HMW‐GS (2*, 5, and 7 subunits) than the early extracted fractions. The trend in the amounts of 2*, 5, and 7 subunits in each fraction from low to high matched the extraction solvent sequence containing from lower to higher levels of HCl. The influence of glutenin protein fractions from the extra‐strong mixing cultivar, Glenlea, on the breadmaking quality of the weak HRSW, McVey, was assessed by enriching (by 1%) the McVey base flour with isolated glutenin protein fractions from Glenlea. The mixograph peak development times and loaf volumes of enriched flour were measured in an optimized baking test. The results indicated that the higher content in Glenlea glutenin of HMW‐GS with higher molecular weight, such as 2*, 5, and 7, seem to be the critical factor responsible for the strong mixing properties of Glenlea. Our results confirmed that subunit 7 occurred in the highest quantity of all the HMW‐GS. Therefore, it seems that the greater the content of larger molecular weight glutenin subunits, the larger the glutenin polymers and the stronger the flour.     

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.     

Differing Effects of Mechanical Dough Development and Sheeting Development Methods on Aggregated Glutenin Proteins

Dough development using sheeting and mechanical dough development (MDD) were compared with respect to the effect the mixing method had on the molecular size distribution and degree of protein thiol exposure of the aggregated glutenin proteins. Although sheeting imparts a lower rate of work input on doughs than does MDD mixing, changes in protein aggregation patterns during mixing were similar for both methods of dough development, indicating that protein disaggregation was important in the process of dough development. In both systems, a reduced rate of change in the protein aggregation patterns was associated with optimum dough development. The MDD mixing was characterized by increasing exposure of the thiol groups on the SDS‐insoluble glutenin during mixing while the sheeting process resulted in fewer exposed thiol groups on both SDS‐soluble and SDS‐insoluble glutenin proteins. This suggested that disulfide bond rupture may not be a required process in dough development and that high effective stresses per se may not be required to develop doughs. This is consistent with a model for dough development that does not require extensive covalent bond rupture but instead involves mainly rupture and reformation of noncovalent interactions such as hydrophobic bonds and hydrogen bonds between protein chains.     

Changes in SDS Solubility of Glutenin Polymers During Dough Mixing and Resting

An online coupling of high‐performance size‐exclusion chromatography (HPSEC) combined with multiangle laser‐light scattering (MALLS) and a reverse‐phase HPLC procedure were used to characterize and reveal the polydispersity of the glutenin polymers of doughs during mixing and resting. Experiments involved doughs prepared from several samples of a common French wheat cultivar (Soissons) differing in total amount of SDS‐unextractable glutenin polymers. During dough mixing, the amounts, size distribution of protein, and glutenin subunit composition within the SDS‐unextractable polymers changed. However, the major changes in SDS‐unextractable glutenin content and size distribution occurred before the peak mixing time (MT) was reached, whereas detectable changes in subunit composition also occurred after the peak MT. Even if sonication, which was used to solubilize the total wheat glutenin, can narrow the glutenin size distribution, HPSEC‐MALLS revealed a close relationship between the SDS solubility of the glutenin polymers and size distribution, confirming a depolymerization and repolymerization hypothesis. During the depolymerization of the SDS‐unextractable polymers, glutenin subunits were released in nonrandom order, which indicated that the polymers have a hierarchical structure. Some HMW glutenin subunits (HMW‐GS), especially 1D×5, were particularly resistant to the depolymerization mechanism. This suggested that the subunit plays a major role in forming the backbone of the SDS‐unextractable polymers, consistent with the potential to form branched structure. These studies suggest that the SDS‐unextrac‐table polymers in flours have a well‐ordered structure that can be modified by dough mixing and resting.     

Preparative Method for In Vitro Production of Functional Polymers from Glutenin Subunits of Wheat

An in vitro method for preparative‐scale production of artificial glutenin polymers utilizes a controlled environment for the oxidation of glutenin subunits (GS) isolated from wheat flour to achieve high polymerization efficiency. The functionality of in vitro polymers was tested in a 2‐g model dough system and was related to the treatment of the proteins before, during, and after in vitro polymerization. When added as the only polymeric component in a reconstituted model dough (built up from gliadin, water solubles, and starch fractions), in vitro polymers could mimic the behavior of native glutenin, demonstrating properties of dough development and breakdown. Manipulating the high molecular weight (HMW)‐GS to a low molecular weight (LMW)‐GS ratio altered the molecular weight distribution of in vitro polymers. In functional studies using the 2‐g mixograph, simple doughs built up from homopolymers of HMW‐GS were stronger than those using homopolymers of LMW‐GS. These differences may be accounted for, at least in part, by different polymer size distributions. The ability to control the size and composition of glutenin polymers shows the potential of this approach for investigating the effects of glutenin polymer size on dough function and flour end‐use quality.