Relationship Between Solvent Retention Capacity and Protein Molecular Weight Distribution,Quality Characteristics,and Breadmaking Functionality of Hard Red Spring Wheat Flour

This research aims to investigate the relationship between the solvent retention capacity (SRC) test and quality assessment of hard red spring (HRS) wheat flour samples obtained from 10 HRS cultivars grown at six locations in North Dakota. The SRC values were significantly (P < 0.05) correlated with flour chemical components (protein, gluten, starch, and damaged starch contents, except arabinoxylan); with farinograph parameters (stability [FST], water absorption, peak time [FPT], and quality number); and with breadmaking parameters (baking water absorption [BWA], bread loaf volume [BLV], and symmetry). Differences in locations and cultivars contributed significantly to variation in quality parameters and SRC values. Suitability of SRC parameters for discriminatory analysis of HRS wheat flour is greatly influenced by molecular weight distribution (MWD) of SDS‐unextractable proteins. SRC parameters, except for sucrose SRC, showed significant (P < 0.01) and positive correlations with high‐molecular‐weight (HMW) polymeric proteins in SDS‐unextractable fractions, whereas only lactic acid SRC exhibited significant (P < 0.01) correlations with low‐molecular‐weight polymeric proteins. HMW polymeric proteins also exhibited positive associations with FPT, FST, BWA, and BLV. The discrepant variation in association of SRC parameters with respect to MWD of SDS‐unextractable proteins could improve segregation of HRS wheat flour samples for quality.     

Simplified Dilute Acetic Acid Based Extraction Procedure for Fractionation and Analysis of Wheat Flour Protein by Size Exclusion HPLC and Flow Field‐Flow Fractionation

A simple, highly efficient and reproducible two‐step extraction procedure using dilute acetic acid without (AN) and then with sonication (AS) has been developed for the fractionation of wheat flour protein. Approximately 97% of total protein was extracted from a Canadian hard red spring wheat flour; an additional 1.2% protein could be recovered by further extraction with 1% DDT and 50% 1‐propanol (AR). Size‐exclusion HPLC (SE‐HPLC) and flow field‐flow fractionation (flow FFF) showed that the AN extract, which accounted for most of the total extractable protein (AN + AS + AR), consisted primarily of monomeric protein. The AS extract was composed primarily of polymeric proteins. Flow FFF showed that AN polymeric protein, including that eluting at the SE‐HPLC void volume, showed smaller Stokes diameters than AS polymeric protein. Flow FFF profiles of AS SE‐HPLC subfractions showed that the void volume subfraction contained monomeric and small polymeric protein in addition to large polymeric protein, indicating formation of larger complexes through interaction between some or all of the components. AN and AS extracts, as well as SE‐HPLC and flow FFF fractions thereof, showed a fairly wide range of values among 12 Canadian hard red and white spring wheat cultivars. The proportion of total protein in the AS extract and in the larger sized polymeric protein fractions from SE‐HPLC and flow FFF were highly positively correlated to farinograph mixing time.     

Molecular Weight Distribution of Wheat Proteins

The molecular weight distribution (MWD) of wheat proteins is becoming recognized as the main determinant of physical dough properties. Studies of high polymers have shown that properties such as tensile strength are related to a fraction of polymer with molecular weight above a critical value and the MWD of this fraction. Elongation to break is treated as a kinetic process with energies of activation for breaking noncovalent bonds and for chain slippage through entanglements. These considerations are related to tensile properties of wheat flour doughs such as those measured by the extensigraph. The MWD of wheat proteins is determined by the relative amounts of monomeric and polymeric proteins and the MWD of the polymeric proteins. The latter, in turn, depends on the ratio of high molecular weight glutenin subunits (HMW-GS) to low molecular weight glutenin subunits (LMW-GS), the specific HMW-GS that result from allelic variation, and the presence of modified gliadins that act as chain terminators. The role of these compositional variables in determining dough extensional properties is discussed in terms of present knowledge. Determination of MWD of wheat proteins is hindered by the difficulty of their solubilization and the lack of methods for reliably measuring very high molecular weights. Among the promising techniques for achieving these measurements are multiangle laser light scattering (MALLS) and field flow fractionation (FFF).     

Use of Size‐Exclusion High‐Performance Liquid Chromatography for Wheat Quality Prediction in Ethiopia

Wheat quality testing facilities in Ethiopia are limited. The aim of this study was to determine whether size‐exclusion high‐performance liquid chromatography (SE‐HPLC) could be used in breeding programs for quality testing. Thirteen Ethiopian and two South African wheat cultivars were evaluated in two diverse environments for milling and dough characteristics. SE‐HPLC was done on the same samples. Across environments, both SDS‐soluble and SDS‐insoluble polymeric proteins significantly influenced important quality characteristics such as SDS‐sedimentation and mixograph development time. The large monomeric proteins, which are mainly gliadins, had a consistently significantly negative effect on quality. The increase of polymeric protein as opposed to monomeric protein led to improvement of quality characteristics. The SDS‐soluble and SDS‐insoluble polymeric proteins were equally important in quality prediction. The amount of polymeric proteins was significantly higher in the high‐protein environment. Despite a large environmental effect on most fractions, a large ratio of polymeric proteins to monomeric proteins (both SDS‐soluble and SDS‐insoluble) can be a good indicator of baking quality. SE‐HPLC is therefore an option to use in breeding programs in Ethiopia for quality evaluation.     

Molecular Characterization of Corn Starch Using an Aqueous HPSEC‐MALLS‐RI System Under Various Dissolution and Analytical Conditions

Molecular characteristics based on absolute weight‐average molecular weight (M_w) and z‐average radius of gyration (R_g) of normal corn starch were analyzed by high‐performance size‐exclusion chromatography (HPSEC) attached to multiangle laser‐light scattering (MALLS) and refractive index (RI) detectors under different starch dissolution and analytical conditions. Autoclaving (121°C, 20 min) or microwave heating (35 sec) provided better HPSEC recovery and higher M_w for starch molecules than simple dissolution in hot water. The M_w for the autoclaved corn amylopectin and amylose fractions separated with a TSK G5,000 column at 60°C were 201 × 10⁶ and 3.3 × 10⁶, respectively. The specific volume for gyration (SV_g) calculated from M_w and R_g could be used for the comparison of molecular compactness which was inversely related to the degree of branching. The SV_g values of amylopectin and amylose fractions in the chromatogram (TSK G5,000, autoclaved for 20 min) were 0.092 and 0.529, respectively. But a portion (20–30%) of large amylopectin molecules did not pass the injection membrane filter (3.0 μm) and the SEC column, resulting in incomplete recovery. The unfiltered portion varied according to the dissolution treatment. Homogenization (7,000 rpm, 5 or 10 min) of the starch solution improved the recovery of the amylopectin fraction, but significantly increased the M_w of the amylose fraction (17 × 10⁶). Sonication for 5 min degraded starch molecules. For accurate analysis of a native starch using an aqueous SEC, the starch should be fully dissolved with proper treatment such as autoclaving or microwaving, and the column should be improved for full recovery of large amylopectin molecules.     

Distribution of Protein Composition in Bread Wheat Flour Mill Streams and Relationship to Breadmaking Quality

Wheat protein quantity and composition are important parameters for wheat baking quality. The objective of this study was to use fractionation techniques to separate the proteins of flour mill streams into various protein fractions, to examine the distribution of these protein fractions, and to establish a relationship between protein composition and breadmaking quality. Nine break streams, nine reduction streams, and three patent flours obtained from three samples of Nekota (a hard red winter wheat) were used in this study. A solution of 0.3M NaI + 7.5% 1-propanol was used to separate flour protein into monomeric and polymeric proteins. The protein fractions, including gliadin, albumin+globulin, HMW-GS, and LMW-GS, were precipitated with 0.1M NH₄Ac-MeOH or acetone. The fractions were statistically analyzed for their distribution in the mill streams. The quantities of total flour protein and protein fractions in flour were significantly different among mill streams. The ratio of polymeric to monomeric proteins in break streams was significantly greater than in the reduction streams. The relationship between protein composition and breadmaking quality showed that the quantities of total flour protein, albumin+ globulin, HMW-GS, and LMW-GS in flour were significantly and positively correlated with loaf volume. The ratio of HMW-GS to LMW-GS had little association with loaf volume. The gliadin content in total flour protein was negatively and significantly correlated with loaf volume. These results indicated that the quantity and composition of protein among the mill streams was different, and this resulted in differences in breadmaking quality.     

Reoxidation Behavior of Wheat and Rye Glutelin Subunits

The ability of HMW and LMW subunits of wheat glutelin to form a polymeric gluten network by intermolecular disulfide bonds is responsible for the unique rheological properties and baking quality of wheat dough. Because the mechanism of gluten formation is not fully understood, the reoxidation behavior of HMW and LMW subunits of wheat glutelin and HMW subunits of rye glutelin was studied. The subunits were isolated from wheat flour cv. Rektor (REK) and from rye flour cv. Danko (DAN) with a selective extraction and precipitation method. For reoxidation, different oxidants (KBrO₃ and KIO₃), protein concentrations (0.5, 1.0, and 2.0%), solvent compositions, pH values (2.0 and 8.0), and reaction times (0–360 min) were compared. The characterization of reoxidized products was achieved by the determination of the thiol content with the Ellman's reagent, and of the M_r distribution by gel‐permeation chromatography. The results demonstrated that both HMW and LMW subunits could be slowly reoxidized with KBrO₃ to polymers with M_r up to several millions. Yield and M_r distribution of polymers were dependent both on the protein concentration and on the molar ratio of oxidants to thiol groups. The HMW subunits of wheat glutelin (HMW‐REK) yielded slightly higher quantities of polymeric proteins than did the HMW subunits of rye (HMW‐DAN). Reoxidation with KIO₃ proceeded much faster than with KBrO₃ and led to lower proportions of polymerized proteins for HMW‐REK and HMW‐DAN. Obviously, more intra‐ and fewer intermolecular disulfide bonds were formed by reoxidation with KIO₃ compared with KBrO₃. In contrast, LMW‐REK was reoxidized with KIO₃ to higher amounts of polymeric aggregates, which indicated that LMW subunits formed intermolecular disulfide bonds with both KIO₃ and KBrO₃. Independent of the protein type and the oxidant used for reoxidation, more inter‐ and fewer intramolecular disulfide bonds were formed when the protein concentration was increased. Single subunits 5, 7, and 10 were isolated from HMW‐REK by preparative acid‐PAGE and were reoxidized with KBrO₃ for 360 min. The M_r distribution indicated that x‐type subunit 5 had a greater tendency to form polymers than x‐type subunit 7. The y‐type subunit 10 was characterized by a lower proportion of polymers after reoxidation than x‐type subunits 5 and 7.     

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

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

Effects of Kernel Vitreousness and Protein Level on Protein Molecular Weight Distribution,Milling Quality,and Breadmaking Quality in Hard Red Spring Wheat

Dark, hard, and vitreous kernel content is an important grading characteristic for hard red spring (HRS) wheat in the United States. This research investigated the associations of kernel vitreousness (KV) and protein content (PC) levels with protein molecular weight distribution (MWD), milling quality, and breadmaking quality characteristics of HRS wheat. The U.S. regional crop quality survey samples from three consecutive growing years were combined into three composite samples with different levels of wheat PC and then further segregated into separate samples with three different levels of KV. Analysis of variance showed that KV level had significant (P < 0.001) effect on variation in test weight, break flour yield, and damaged starch content. Among protein MWD parameters analyzed by size‐exclusion HPLC, the high‐molecular‐weight polymeric proteins in the SDS‐unextractable fraction had significant (P < 0.01) association with KV. Regression analysis indicated that addition of KV to the PC level improved the model for both farinograph and baking water absorption values in all three growing years. This information could help the flour milling and baking industries to further segregate HRS wheat based on KV levels in addition to PC levels for their intended end‐use applications.     

Variation and Correlation of Protein Molecular Weight Distribution and Semolina Quality Parameters for Durum Genotypes Grown in North Dakota

This research assessed variation of protein molecular weight distribution (MWD) parameters and their correlations with quality characteristics of semolina samples that were obtained from durum genotypes grown in North Dakota. Sodium dodecyl sulfate buffer extractable and unextractable proteins in semolina were analyzed for MWD by size‐exclusion HPLC with a microbore column. ANOVA indicated that quantitative variations of all the HPLC protein fractions were significantly (P < 0.001) influenced by growing environments. The extractable and unextractable gluten proteins correlated differently with semolina gluten characteristics. Both gluten index and mixograph classification showed positive correlations (P < 0.05) with unextractable polymeric proteins and negative correlations (P < 0.05) with extractable gliadins and polymeric proteins. Quantitative variations of gluten proteins greatly influenced spaghetti cooking characteristics. Specifically, cooked spaghetti firmness (CSF) had high and positive simple linear correlations (P < 0.001) with quantity of gluten proteins in both extractable and unextractable fractions. However, a qualitative MWD parameter, percentage of the extractable gliadins in total protein, had a negative genotypic correlation with CSF (r = –0.81, P < 0.01), whereas percentage of the unextractable polymeric proteins had a positive genotypic correlation (r = 0.75, P < 0.01). Those two MWD parameters also showed significant (P < 0.05) variations for genotypes, indicating that they might be useful for screening durum genotypes for pasta cooking quality.     

Separation and Characterization of Barley Starch Polymers by a Flow Field‐Flow Fractionation Technique in Combination with Multiangle Light Scattering and Differential Refractive Index Detection

Flow field‐flow fractionation (flow FFF) with frit inlet and frit outlet mode (FIFO) was coupled online to multiangle light scattering (MALS) and refractive index (RI) detectors to investigate the molecular characteristics of normal and zero amylose barley starch polymers. Application of two different cross‐flows, 0.35 mL/min followed by 0.1 mL/min, and constant channel and frit flows of 0.1 and 1.0 mL/min, respectively, permitted a complete separation of amylose and amylopectin. The improved signals from the detectors due to application of the FIFO mode enabled the proper characterization of the small molecular weight species, as well as significantly enhanced the reproducibility of the measurements. The weight‐average molecular weight (M_w) and zaverage root‐mean‐square (RMS) radii of gyration (R_g) values for amylose and amylopectin in the normal starch samples were 2.3 × 10⁶ and 280 × 10⁶, and 107 and 260 nm, respectively. The M_w and R_g of amylopectin in the zero amylose starch samples were 360 × 10⁶ and 267 nm, respectively. The slopes (α) obtained by plotting log M_w versus log R_g for amylose and amylopectin were 0.6 and 0.3, respectively. These results are in good agreement with the theoretical prediction of the molecular conformation of amylose and amylopectin.     

Starch Molecular Mass and Size by Size-Exclusion Chromatography in DMSO-LiBr Coupled with Multiple Angle Laser Light Scattering

The weight average molar mass (M_w) and root mean square radii of starches from waxy maize (Amioca), waxy rice flour, cassava, Hylon V, Hylon VII, and potato amylose were determined by size-exclusion chromatography (SEC) and multiple-angle laser light scattering (MALLS). Dimethylsulfoxide (DMSO) containing 50 mM LiBr was used to dissolve the starches and also served as the mobile phase. SEC with large particle size polystyrene divinylbenzene packing materials and MALLS instrumentation were evaluated for the ability to separate and determine molar mass (MM) of starch polymers, respectively. The determination of M_w by MALLS is necessary because the M_w of many cereal starches exceeds the available molecular standards by one or two orders of magnitude. The M_w depends on the method of calculation. The M_w (Berry method) of starch from waxy corn was 2.27 × 10⁸ Da, waxy rice 8.9 × 10⁷ Da, cassava 5.7 × 10⁷ Da, Hylon V 2.7 × 10⁷ Da, Hylon VII 4.8 × 10⁶ Da, and potato amylose 1.9 × 10⁵ Da. Recovery dropped dramatically for molecules with root mean square radii >200 nm.     

Salt‐resistant and salt‐sensitive wheat genotypes show similar biochemical reaction at protein level in the first phase of salt stress

Salinity has a two‐phase effect on plant growth, an osmotic effect due to salts in the outside solution and ion toxicity in a second phase due to salt build‐up in transpiring leaves. To elucidate salt‐resistance mechanisms in the first phase of salt stress, we studied the biochemical reaction of salt‐resistant and salt‐sensitive wheat (Triticum aestivum L.) genotypes at protein level after 10 d exposure to 125 mM–NaCl salinity (first phase of salt stress) and the variation of salt resistance among the genotypes after 30 d exposure to 125 mM–NaCl salinity (second phase of salt stress) in solution culture experiments in a growth chamber. The three genotypes differed significantly in absolute and relative shoot and root dry weights after 30 d exposure to NaCl salinity. SARC‐1 produced the maximum and 7‐Cerros the minimum shoot dry weights under salinity relative to control. A highly significant negative correlation (r² = –0.99) was observed between salt resistance (% shoot dry weight under salinity relative to control) and shoot Na⁺ concentration of the wheat genotypes studied. However, the salt‐resistant and salt‐sensitive genotypes showed a similar biochemical reaction at the level of proteins after 10 d exposure to 125 mM NaCl. In both genotypes, the expression of more than 50% proteins was changed, but the difference between the genotypes in various categories of protein change (up‐regulated, down‐regulated, disappeared, and new‐appeared) was only 1%–8%. It is concluded that the initial biochemical reaction to salinity at protein level in wheat is an unspecific response and not a specific adaptation to salinity.     

A Novel Pair of HMW Glutenin Subunits from Aegilops searsii Improves Quality of Hexaploid Wheat

This study involved screening of wild species of wheat in search of functionally useful seed storage proteins for improvement of breadmaking quality of wheat (Triticum aestivum). After screening of 177 disomic addition lines (DALs) of wheat belonging to different wild species, Aegilops searsii DALs were selected and studied in detail. These DALs of Ae. searsii were from chromosome 1S^s to 7S^s in the background of cultivated wheat cv. Chinese Spring (CS). By analyzing these addition lines, genetic loci of actively expressed genes for the high molecular weight glutenin subunits (HMW‐GS) and gliadin were found on the chromosome 1S^s for the first time and have been designated as Glu‐S^s1 and Gli‐S^s1, respectively. Disomic addition line of chromosome 1S^s (DAL1S^s) showed improved dough strength in different generations compared with CS. SDS sedimentation value and specific sedimentation of DAL1S^s were higher than CS. Mixograph peak height and band width were higher, with no difference in mixing peak time from CS. All these factors indicate a positive effect of quantity as well as quality of gluten proteins of Ae. searsii. This was further supported by increased polymer formation in DAL1S^s because the ratio of unextractable polymeric protein to total polymeric protein (UPP/TPP%) of DAL1S^s was significantly higher than CS. Genes for HMW‐GS (major determinant of end‐product quality in wheat) of Ae. searsii were cloned and sequenced from the DAL1S^s. Phylogenetic analysis of deduced amino acid sequences showed that both x and y HMW‐GS were more similar to that of D genome rather than B genome of wheat. Although S genome is structurally more similar to B genome of wheat, functionally it is more similar to the D genome of wheat and possesses good quality HMW‐GS required for improvement of breadmaking quality of wheat.     

Effect of l‐Cysteine on the Rheological Properties of Wheat Flour

Extrudate expansion of cereal‐based products is largely dependent on the molecular interactions and structural transformations that proteins undergo during extrusion processing. Such changes strongly influence the characteristic rheological properties of the melt. It is possible to modify rheological properties of wheat flour during extrusion processing, in particular shear viscosity, with cysteine. The objective of this work was to further develop an understanding of the molecular interactions and structural transformations of wheat flour from dynamic oscillatory rheological measurements. Temperature and frequency sweeps were conducted in the linear viscoelastic range of the material. Changes in the storage modulus (G′), the loss modulus (G″) and the loss tangent (tan δ) of 25% moisture wheat flour disks as a function of cysteine concentration (0–0.75%) were monitored. Molecular weight between cross‐links (M_c) and the number of cross‐links (N_c) per glutenin molecule were determined from frequency sweep data. Increasing cysteine concentration broke cross‐links by decreasing G′ maximum and increasing tan δ values. Molecular weight between cross‐links increased and the number of cross‐links decreased. G′ values from temperature sweeps showed a similar trend. This information leads to a better understanding of the viscoelastic behavior of wheat flour doughs during extrusion cooking and elucidation of protein‐protein reaction mechanisms and other interactions in extruded cereal‐based snack foods.     

Molecular Weight Distribution of Proteins in Hard Red Spring Wheat: Relationship to Quality Parameters and Intrasample Uniformity

Molecular weight distribution (MWD) of proteins extracted from hard red spring wheat was analyzed by size‐exclusion HPLC to investigate associations with wheat and breadmaking quality characteristics. Certain protein fractions were related to associations between wheat and breadmaking parameters, specifically when effect of quantitative variation of protein on those parameters was statistically eliminated by partial correlation analysis. SDS‐unextractable high molecular weight polymeric proteins had positive partial correlations with percent vitreous kernel content and breadmaking parameters, including mix time and bread loaf volume. SDS‐extractable protein fractions that were eluted before the primary gliadin peak had positive partial correlations with kernel hardness and water absorption parameters. The proportion of main gliadin fractions in total protein had a negative partial correlation with bread loaf volume and positive correlations with kernel hardness and water absorption parameters. Intrasample uniformity in protein MWD and kernel characteristics was estimated from three kernel subsamples that were separated according to single kernel protein content within individual wheat samples by a single‐kernel near‐infrared sorter. Wheat subsamples were significantly different in protein MWD. Intrasample uniformity in protein MWD did not differ greatly among wheat samples.     

Separation of Water‐Soluble Proteins from Cereals by High‐Performance Capillary Electrophoresis (HPCE)

Most research concerning grain proteins has concentrated on the gluten storage proteins. The albumins and globulins are the water‐ and salt‐soluble proteins that contain biologically active enzymes and enzyme inhibitors. A free‐zone capillary electrophoresis method was developed to separate these proteins. Optimization included sample extraction method, capillary temperature, buffer composition, and additives. The optimal conditions for separation of these proteins was 50 μm i.d. × 27 cm (20 cm to detector) capillary at 10 kV (with a 0.17 min ramp‐up time) and 25°C. The optimum buffer was 50 mM sodium phosphate, pH 2.5 + 20% acetonitrile (v/v) (ACN) + 0.05% (w/v) hydroxypropylmethyl‐cellulose (HPMC) + 50 mM hexane sulfonic acid (HSA). Sample stability was an issue that was addressed by lyophilizing fresh extracts and redissolving in aqueous 50% ethylene glycol and 10% separation buffer. This method was successfully used in both wheat flour and whole meal samples. Comparisons were made of several wheats of different classes as well as several cereal grains. This methodology could be useful in screening cereal grains for important enzymes and their impact on end‐use quality such as food functionality, food coloration, and malting quality.     

Effect of Aelia spp. and Eurygaster spp. Damage on Wheat Proteins

The effect of Aelia spp. and Eurygaster spp. wheat bugs on the protein fractions of different wheat cultivars has been studied by size‐exclusion high‐performance liquid chromatography (SE‐HPLC) and free‐zone capillary electrophoresis (FZCE). Those methods were used to quantify and characterize the extent of protein modification. A decrease in the amount of alcohol‐insoluble polymeric proteins along with an increase in the alcohol‐soluble polymeric proteins and gliadins were observed in damaged wheat. The high molecular weight (HMW) and low molecular weight (LMW) glutenin fractions were barely detected in the incubated damaged wheat from some cultivars, which indicated hydrolysis of those proteins by the bug proteinases. In damaged wheats, both incubated and unincubated, gliadin electrophoregrams revealed the presence of some new peaks with mobilities similar to the ω gliadins. The overall results suggest that the bug proteinases are potent enzymes that appear to be nonspecific because they hydrolyze all gluten proteins.     

Quantitation of LMW‐GS to HMW‐GS Ratio in Wheat Flours

A comparison was made of methods for measuring the LMW/HMW glutenin subunit (GS) ratio for glutenin. A set of near‐isogenic wheat lines with the number of HMW‐GS varying from 0 to 5 was utilized to provide a wide range of LMW/HMW‐GS. Glutenin preparations were obtained from ground whole meal after solubilization of monomeric proteins by dimethyl sulfoxide (DMSO) or 50% propanol or by fraction collection from a preparative SE‐HPLC column. Analyses were made on the reduced glutenin from each of the three preparations by RP‐HPLC, SE‐HPLC, and SDS‐PAGE. Both solvents, DMSO and 50% propanol, extracted appreciable amounts of polymeric protein, thus casting some doubts on the accuracy of the determinations. This problem was largely avoided when the polymeric fraction was collected from the eluate of a total glutenin extract run on a preparative SE‐HPLC column. Less glutenin was removed by the two solvents for lines with a greater number of HMW‐GS or with strength‐associated HMW‐GS 5+10 coded by the 1D chromosome. Collection of the polymeric protein in SE‐HPLC, followed by separation of the glutenin subunits in RP‐HPLC, was the best method for quantitating the LMW/HMW‐GS ratio. SE‐HPLC gave a clear separation of the two groups of subunits as well as HMW albumins. RP‐HPLC has the potential advantage of being able to quantitate individual subunits.     

Association Between % SDS‐Unextractable Polymeric Protein (%UPP) and End‐Use Quality in Chinese Bread Wheat Cultivars

Trial I, with 33 spring cultivars, and trial II, with 21 winter cultivars, sown in four environments in the northwestern China spring wheat region and northern winter wheat region, respectively, were used to study the effect of genotype and environment on the size distribution of polymeric proteins. Association between quantity and size distribution of polymeric protein and dough properties (both trials) and northern‐style Chinese steamed‐bread (CSB) (trial I) and pan bread (trial II) qualities were also investigated. In trial I, all protein attributes, such as flour protein content, SDS‐extractable polymeric protein in the flour (EPP), SDS‐unextractable polymeric protein in the flour (UPP), and percent UPP in total polymeric protein (%UPP), were largely determined by environment, whereas variation in dough strength resulted from variation in UPP and %UPP across environments. In trial II, EPP was largely determined by environment, and UPP and %UPP were largely determined by genotype. These differences might result from different levels of protein content and dough strength in the two trials. The EPP was positively correlated with dough extensibility and was generally negatively correlated with dough stability and maximum resistance in both trials. However, %UPP was significantly positively correlated with dough stability and maximum resistance and end‐use quality in both trials. In trial I, correlation coefficients between %UPP and maximum resistance and CSB score were r = 0.90 and 0.71, respectively, whereas in trial II, the correlation coefficients between %UPP and maximum resistance and pan bread score were 0.96 and 0.87, respectively. Therefore, selection for high %UPP together with high‐quality glutenin subunits should lead to improved dough strength and end‐use quality in Chinese wheats.