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.     

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.     

Relationship between endogenous protein disulfide isomerase family proteins and glutenin macropolymer

The effects of endogenous protein disulfide isomerase (PDI) family proteins on the properties of gluten proteins in dough during breadmaking were determined using bacitracin, an inhibitor of PDI. Bread loaf volume in the presence of bacitracin was increased to 118% of that in the absence of bacitracin. The addition of bacitracin caused a decrease in the extension tolerance of the dough. The amount of sodium dodecyl sulfate (SDS)-insoluble glutenin macropolymer (GMP) in dough decreased to approximately 70% of that in flour during the 20 min of mixing for doughmaking. The addition of bacitracin to dough caused a dramatic GMP decrease, corresponding to ～20-30% of that in flour during the 20 min of mixing. The decrease in GMP was compensated by an increase in SDS-soluble glutenin polymer. Taken together, these results suggest that the endogenous PDI family proteins in flour suppress the depolymerization of GMP during dough mixing.     

Protein and Quality Characterization of Triticale Translocation Lines in Breadmaking

Introduction of high molecular weight glutenin subunits (HMW‐GS) from the Glu‐D1d locus of wheat into triticale restores the genetic constitution of high molecular weight glutenin loci to that of wheat and subsequently improves the breadmaking quality of triticale. One means of achieving such restoration of the genetic constitution is through the use of translocation lines. The aim of this study was to evaluate and compare the performance of translocations 1A.1D and 1R.1D with HMW‐GS 5+10 and 2+12 in terms of physical dough tests and baking quality using four different sets of triticale lines, GDS7, Trim, Rhino, and Rigel. In general, significantly lower milling quality (flour yield), very low mixing times with lower loaf volume were typical of all the triticales studied except 1A.1D 5+10 lines, when compared to hard wheat flour (Pegaso). Among the lines studied, significantly higher loaf volume, mixograph dough development time (MDDT), and maximum resistance to extension (R_max) were observed with 1A.1D 5+10 lines indicating that translocation of the Glu‐D1d allele with HMW‐GS 5+10 was beneficial in terms of improving the quality attributes. Although pure triticale flour from these lines did not possess the functional characteristics for good quality bread, the translocation 1A.1D that contains HMW glutenin subunits 5+10 showed significant improvement in quality characteristics, and could reasonably be expected to yield commercially satisfactory bread loaves when combined with bread wheat flour. Significantly higher UPP, R_max, and MDDT values along with a lower gliadin‐to‐glutenin ratio in 1A.1D 5+10 of GDS7 and Rigel sets indicate that the molecular weight distribution was shifted to higher molecular weights, resulting in greater dough strength associated with 5+10 subunits.     

Rheological Properties of Full-Formula Doughs Derived from Near-Isogenic 1BL/1RS Translocation Lines

Four pairs of near-isogenic wheat lines, with and without the 1BL/1RS translocation, and differing at the Glu-1 loci (coding for high molecular weight [HMW] glutenin subunits) were evaluated for their dough mixing properties, dough stickiness, and baking performance. In all 1BL/1RS translocation lines, weakening of the dough consistency occurred within 2 min past peak time. The full-formula dough from every 1BL/1RS translocation line exhibited poor dough mixing characteristics and increased stickiness compared to the corresponding wheat control. The HMW glutenin subunits coded by the Glu-A1 locus had no apparent effect on mixing properties, but did have a slight effect on the dough stickiness at two of the four stages of dough mixing. Glu-B1 and Glu-D1 loci encoded glutenin subunits produced significant changes in dough mixing properties and dough stickiness, respectively. With respect to baking performance, there was no significant difference between loaf volumes of 1BL/1RS versus control wheats for three of four near-isogenic pairs. Within the 1RS-group, the translocation lines containing HMW glutenin subunits 5+10 produced bread with greater loaf volumes than the pairs containing its allelic counterpart 2+12. Loaf volume was not influenced by the subunits associated with the Glu-B1 loci. In general, the breads baked from 1BL/1RS translocation lines had a relatively poor crumb and crust quality and contained larger gas cells than the wheat controls. In comparing isogenic pairs, the magnitude of the difference in loaf volume between the control wheat and the corresponding 1BL/1RS translocation line was greater in the pair unique for HMW subunits 5+10; the difference was primarily due to the stronger mixing properties of the wheat control.     

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.     

Transgenic Enhancement of High‐Molecular‐Weight Glutenin Subunit 1Dy10 Concentration: Effects in Wheat Flour Blends and Sponge and Dough Baking

Dough strength is needed for efficient breadmaking quality. This property is strongly influenced in wheat (Triticum aestivum L.) by gluten seed storage proteins and, in particular, by high‐molecular‐weight (HMW) glutenin subunit composition. Experiments were designed to elevate expression of a key native HMW glutenin subunit (1Dy10) via genetic engineering and to determine whether resultant flours can be used in sponge and dough applications, the most common commercial bread‐baking procedure. Both unblended and blended samples from transgenic and nontransgenic sister lines were tested, with blended samples being formed by addition to a control sample. Dough properties, as determined by farinograph evaluation, were improved by the transgene‐encoded increases in 1Dy10 in both undiluted and blended flours. Mean farinograph stability of transgenic samples was twice that of the control, and blends with transgenic samples demonstrated increases in stabilities proportional to the amount of transgenic flour included. Mean farinograph quality numbers of transgenic samples, and of all blends containing transgenic flour, were significantly higher than both the control and all nontransgenic treatments. In the sponge and dough bake procedure, undiluted transgenic samples induced lower scores, relative to both control and undiluted nontransgenic samples, for water absorption, crumb body firmness, and loaf volume. In blends, however, the transgenic samples resulted in improvements in some sponge and dough loaf attributes, including loaf symmetry and crumb color score, without any concomitant loss of loaf volume in transgenic blends. These improved variables relate to finished product appearance and to consumer selection in markets. The use of transgenic flours with increased 1Dy10 glutenin content in commercial blends could provide advantages in sponge and dough bake applications.     

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.     

Intercultivar Variation in the Quantity of Monomeric Proteins,Soluble and Insoluble Glutenin,and Residue Protein in Wheat Flour and Relationships to Breadmaking Quality

A new fractionation procedure based on differential solubility was applied to wheat flour proteins to evaluate the relationship between protein fractions and functionality for breadmaking. Flour was initially extracted with 50% 1-propanol. Monomeric proteins (mainly gliadins) and soluble glutenin contained in the 50% propanol soluble extract were fractionated by selective precipitation of the glutenin by increasing the concentration of 1-propanol to 70%; monomeric proteins remain in the supernatant. Insoluble glutenin in the 50% propanol insoluble residue was extracted using 50% 1-propanol containing 1% dithiothreitol (DTT) at 60°C. Protein in the final residue was extracted using SDS with or without DTT. It comprised mainly Glu-1D high molecular weight glutenin subunits and nongluten polypeptides. For seven Canadian cultivars of diverse breadmaking quality, there was relatively little variation in the percentage of flour protein corresponding to monomeric proteins (48–52%) and residue protein (14–18%). In contrast, intercultivar variation in soluble and insoluble glutenin was substantial, with contents of 10–20% and 12–28% of flour protein, respectively. Soluble and insoluble glutenin were also highly correlated with physical dough properties, accounting for 83–95% of the variation of individual dough rheological parameters (except dough extensibility), and ≈ 74% of the variation in loaf volume. In contrast, monomeric and residue protein fractions were poorly associated with breadmaking quality. However, among the four protein fractions, only residue protein was significantly correlated (r = -0.79) with dough extensibility. The flour sample with the highest and lowest concentrations of insoluble and soluble glutenin, respectively, as well as marginally the lowest concentrations of monomeric and residue proteins was Glenlea, a cultivar of the Canada Western Extra Strong Red Spring wheat class which characteristically possesses distinctly strong dough mixing properties.     

Protein Changes During Various Stages of Breadmaking of Four Spring Wheats: Quantification by Size-Exclusion HPLC

Protein changes for four hard red spring wheat genotypes (Len, Marshall, 215, and Butte 86) were assessed at various stages of breadmaking using a size-exclusion HPLC technique. Breadmaking stages considered were flour, after mixing, before punching, after punching, after fermentation, and after proofing. Quality and functional characteristics of the four wheat genotypes were determined. The three main protein groups isolated by SE-HPLC were further characterized by SDS-PAGE. A direct relationship between polymeric glutenin (peak I of SE-HPLC fractions) in flours and loaf volume was found for the three wheat genotypes with identical high molecular weight glutenin subunit (HMW-GS) composition (2*, 7+9, 5+10) and one line with similar HMW-GS composition (2*, 7+9, 2+12), differing in the Glu-D1 locus. Quantitative changes in the distribution of SDS-soluble proteins fractionated by SE-HPLC were also examined. Peak I proteins (polymeric proteins) from SDS-extractable proteins tend to decrease during breadmaking, while peak III proteins (low molecular weight) tend to increase. Peak II (monomeric proteins, medium molecular weight) did not show a change in quantity during breadmaking. These results seem to indicate that some type of rearrangement took place during the breadmaking process to release proteins of smaller molecular weight.     

Comparative study of the effect of incorporated individual wheat storage proteins on mixing properties of rice and wheat doughs

The aim of this work was to compare the effects of incorporated wheat storage proteins on the functional properties of rice and wheat flours. The advantage of rice as a base flour compared to wheat is that it does not contain any wheat flour components and, therefore, has no interactive effect between wheat glutenin proteins. The incorporation of individual HMW glutenin subunit proteins (Bx6, Bx7, and By8) in different ratios had significant positive effects on the mixing requirements of both rice and wheat doughs. Reconstitution experiments using two x+y type HMW-GS pairs together with a bacterially expressed LMW-GS have been also carried out in this study. The largest effects of polymer formation and mixing properties of rice flour dough were observed when Bx and By subunits were used in a 1:1 ratio and HMW and LMW glutenin subunits in a 1:3 ratio. However, using the same subunit ratios in wheat as the base flour, these synergistic effects were not observed.     

Tyrosine cross-links: molecular basis of gluten structure and function

The formation of the large protein structure known as "gluten" during dough-mixing and bread-making processes is extremely complex. It has been established that a specific subset of the proteins comprising gluten, the glutenin subunits, directly affects dough formation and breadmaking quality. Glutenin subunits have no definitive structural differences that can be directly correlated to their ability to form gluten and affect dough formation or breadmaking quality. Many protein structural studies, as well as mixing and baking studies, have postulated that disulfide bonds are present in the gluten structure and contribute to the process of dough formation through the process of disulfide-sulfhydryl exchange. Evidence presented here indicates that tyrosine bonds form in wheat doughs during the processes of mixing and baking, contributing to the structure of the gluten network. The relative contributions of tyrosine bonds and disulfide--sulfhydryl interchange are discussed.     

Modeling of Dough Mixing Profile Under Thermal and Nonthermal Constraint for Evaluation of Breadmaking Quality of Hard Spring Wheat Flour

This research was initiated to investigate associations between flour breadmaking traits and mixing and empirical dough rheological properties under thermal stress. Thirty hard spring wheat flour samples were analyzed by a Mixolab standard procedure. Mixolab profiles were divided into six different stages, and torque measurements of individual stages were modeled by nonlinear curve fitting using a compound of two solution searching procedures, multidimensional unconstrained nonlinear minimization and genetic algorithm. Mixing patterns followed exponential equations. Dough torque patterns under heat constraint, specifically dough thermal weakening and pasting profiles, were described by a sigmoid logistic equation as a function of time. Dough stability during heating appeared important for bread loaf volume increase from significant correlations between bread loaf volume and parameters generated from models of a dough thermal weakening stage. Multivariate continuum regression was employed to calibrate prediction models of baking traits using Mixolab parameters. Coefficients of determination estimated from prediction models and cross‐validation were greater than 0.98 for bake water absorption, mixing time, and bread loaf volume, indicating that the Mixolab parameters have a potential to enhance evaluation of flour breadmaking quality.     

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

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

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

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

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.     

Characterization of Monomeric and Glutenin Polymeric Proteins of Hard Red Spring Wheats During Grain Development by Multistacking SDS-PAGE and Capillary Zone Electrophoresis

Three cultivars of hard red spring (HRS) wheats with identical high molecular weight (HMW) glutenin subunit composition (5+10 type, Glu-D1d) but different dough properties and breadmaking quality were used in this study. The synthesis and accumulation characteristics of different protein fractions during grain development were examined. Samples were collected at three-day intervals from anthesis to maturity between day 10 to day 37. The nonreduced SDS-extractable glutenin aggregates of developing grains were characterized by a multistacking SDS-PAGE procedure to obtain information on the size distribution and polymerization of glutenin aggregates. The HMW to low molecular weight (LMW) glutenin subunit ratio was determined for its relationship to polymerization of the various glutenin aggregates of different molecular sizes. Glutenin proteins were quantified using an imaging densitometer. In addition, albumins and globulins, α- and β-gliadins, γ-gliadins, and ω-gliadins were separated by capillary zone electrophoresis. The results indicated that albumins-globulins, gliadins, and glutenins in developing grains were present at 10 days after anthesis or earlier. Albumin-globulins decreased in proportion, while gliadins increased in proportion during grain development. Polymerization of glutenin aggregates occurred 10 days after anthesis or earlier and increased significantly throughout the grain-filling period until maturity. Larger aggregates of glutenin increased in proportion, while smaller ones decreased in proportion during grain development. Ratio of polymers to monomers increased significantly from day 10 to day 22 of grain development and then remained constant until grain maturity. Glutenin polymers arrived at their maximum in proportion to total SDS-extractable proteins or monomers at day 22 after anthesis while the molecular size of these polymers continued to increase, as indicated by a rapid increase in proportion of HMW to LMW glutenin subunits. Significant differences were found in accumulation rates of glutenin polymers among the three cultivars. Cultivars Kulm and Grandin, with better breadmaking quality, appeared to have greater rates of accumulation and HMW subunit synthesis or formation of larger polymers than did Sharp, a cultivar with poorer quality. Significant differences were found among the three cultivars in the proportion of albumins-globulins and gliadins during grain development. However, no significant differences were found among the cultivars in the proportion of albumins-globulins, α-, β-, γ-, and ω-gliadins at grain maturity. Varietal differences in breadmaking quality were due mainly to the differences in glutenin polymers such as ratio of polymeric to monomeric proteins, molecular size distribution, and ratio of HMW to LMW glutenin subunits among wheat cultivars of 2*, 7+9, and 5+10 subunit types. The better breadmaking cultivars might be characterized with higher proportions of glutenins and greater proportion of HMW subunits in total SDS-extractable proteins than the poorer quality cultivar. However, more genotypes need to be examined.     

Depolymerization of the Glutenin Macropolymer During Dough Mixing: I. Changes in Levels,Molecular Weight Distribution,and Overall Composition

Changes in the amounts, molecular weight distributions, and levels of major groups of subunits in the glutenin macropolymer (GMP) of doughs during mixing were investigated. The GMP (gel protein) is the unreduced fraction of gluten protein that remains as a layer on top of the starch after extraction of SDS-soluble proteins and centrifugation. Experiments involved doughs prepared from flours derived from one weak and one strong cultivar and lines derived from cv. Olympic that were null for specific high molecular weight glutenin subunits (HMW-GS). During mixing, the amount of GMP decreased; the major changes occurred before peak mixing time (MT, achievement of peak resistance). In addition, the average apparent molecular weight of GMP (determined by both size-exclusion HPLC and multilayer gel electrophoresis) decreased during mixing, but in this case, the major changes were seen later in the mixing process, during dough breakdown. Even after extensive mixing, polymers and oligomers were released, not free glutenin subunits. During dough breakdown, the composition of GMP also changed, such that the proportion of HMW-GS decreased but β-amylases/D low molecular weight glutenin subunits (LMW-GS) increased. Changes in the total amounts of other LMW-GS typically were smaller with a decrease in the proportion of B subunits and an increase in the proportion of C subunits. The major changes in GMP composition were observed after peak MT (peak resistance) occurring earlier and to a greater extent in the weaker dough. Our results suggest that dough breakdown during mixing may be triggered by loss of HMW-GS, leading to changes in the molecular weight distribution and composition of the disulfide-bonded GMP.     

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

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

Effect of Multiple Substitution of Glutenin and Gliadin Proteins on Flour Quality of Canada Prairie Spring Wheat

The effect of genetic substitution of two to four glutenin and gliadin subunits from a Canada Prairie Spring (CPS) cv. Biggar BSR into Alpha 16, another CPS wheat line, was studied for rheological and baking quality. Results from double substitution showed that the presence of a gliadin component from Biggar BSR (BGGL) and low molecular weight glutenin subunit 45 (LMW 45) contributed to improved dough strength characteristics. Presence of BGGL in combination with high molecular weight glutenin subunit 1 (HMW 1) or 17+18 (HMW 17+18) also showed improved dough strength over control Alpha lines. When three or four protein subunits were substituted, even though improved quality performance was observed, it was associated with the negative effect of lowered flour water absorptions in spite of similar protein contents. The study confirms that LMW glutenins, as well as gliadins, play an important role along with HMW glutenins in wheat flour quality. CPS wheat lines with improved dough strength properties can be selected from the double substitution lines with the combination of BGGL/LMW 45 and BGGL/HMW 1.