Effect of Morphology of Mechanically Developed Wheat Flour and Water on Starch from Gluten Separation Using Cold Ethanol Displacement

The mechanical development of wheat flour and water creates micro and macro structures in dough or batter that critically influence the ability to separate starch from protein by fluid displacement. This study sought to identify specific structural and rheological features and to relate these to separation as indexed by the separation factor. Structural features, especially protein and starch distributions, were examined using visible light microscopy applied to dough samples that had been exposed to a protein dye. Flour and water samples were developed in a Brabender microfarinograph at conditions (water content and time of development) generally suitable for use of the USDA Western Regional Research Center, cold‐ethanol fluid‐displacement method. No truly homogenous structures were observed. However, distinct segregation of protein and starch were apparent at all conditions. Structural features correlated qualitatively with the success of separation indexed by the overall separation factor (α_p/s) for the separation process. Highly segregated states characterized by large protein bands, clustered starch, and large open spaces were obtained with intermediate development (25 ± 5 min) and were most readily separated (α_p/s = 118 ± 7). Segregated states with relatively thin protein bands (≤10 μm dia) in complex networks entrapping starch were obtained after additional development (≥45 min) and were less completely separable (α_p/s = 32 ± 2). Segregated states with irregularly organized protein in the form of clumps and bands were obtained with minimal development and were partially separable (α_p/s = 65 ± 4). Consistency indicated on the microfarinograph increases monotonically throughout and beyond the period of maximum separability. However, elasticity changes and a high rate of increase in consistency evident in the microfarinogram may reflect changes in the structure that also reduce separability. The study demonstrated the use of the ethanol method to isolate development from displacement phenomena forindependent study.     

Farinograph Responses for Wheat Flour Dough Fortified with Wheat Gluten Produced by Cold‐Ethanol or Water Displacement of Starch

The objective of this research was to identify and define mixing characteristics of gluten‐fortified flours attributable to differences in the method for producing the gluten. In these studies, a wheat gluten concentrate (W‐gluten) was produced using a conventional process model. This model applied physical water displacement of starch (dispersion and screening steps), freeze‐drying, and milling. W‐gluten was the reference or “vital” gluten in this report. An experimental W‐concentrate was produced using a new process model. The new model applied coldethanol (CE) displacement of starch (dispersion and screening steps), freeze‐drying, and milling. Freeze‐drying was used to eliminate thermal denaturation and thereby focus on functional changes due only to the separation method. The dry gluten concentrates were blended with a weak, low‐protein (9.2%), soft wheat flour and developed with water in a microfarinograph. We found that both water and cold‐ethanol processed gluten successfully increased the stability (St) and improved mixing tolerance index (MTI) to create in the blended flour the appearance of a breadbaking flour. Notably, in the tested range of 9–15% protein, the St for CE‐gluten was always higher then the St for W‐gluten. Furthermore, the marginal increase in St (slope of the linear St vs. protein concentration) for the CE‐gluten was ≈57% greater than that for the W‐gluten. The slope of the MTI vs. protein data was lower for the CE‐gluten by 24%. Flour fortified with CE‐gluten exhibited higher water absorption (up to 1.8% units at 13.5% P) than flour fortified with W‐gluten.     

Effect on Dough Functional Properties of Partial Fractionation,Redistribution, and In Situ Deposition of Wheat Flour Gluten Proteins Exposed to Water,Ethanol, and Aqueous Ethanol

The application of the cold‐ethanol laboratory fractionation method to the bulk separation of wheat starch and gluten is accompanied by incidental dissolution, removal, or redeposition of a small part of the functional gliadin protein. The new distribution resulting from process incidental redeposition of soluble components or by purposeful add‐back of soluble and leached components can lead to differences in functionality and more difficult recovery of native properties. To assess this issue, we exposed several wheat flour types to ethanol and water (50–90% v/v) solutions, water, and absolute ethanol at 22°C and –12°C. The exposure was mass conserving (leached components returned to substrate by evaporation of the solvent without separation of phases) or mass depleting (leached components not returned to substrate). The result of the mass‐conserving contact would be flour with altered protein distributions and intermolecular interactions. The result of the mass‐depleting contact would also include altered protein content. Furthermore, the mass‐conserving contact would model an industrial outcome for a cold‐ethanol process in which leached components would be added back from an alcohol solution. The leaching result was monitored by mixography of the flour, nitrogen analysis, and capillary zone electrophoresis of extracts. Although dough rheology was generally like that of the source flour, there were notable differences. The primary change for mass‐conserving contact was an increase in the time to peak resistance and a decrease in the rate of loss of dough resistance following peak resistance. These changes were in direct proportion to the amount of protein mobilized by the solvent. Leaching at 22°C, prevented dough formation for most aqueous ethanol concentrations and greatly reduced gliadin protein content. Minimal changes were noted for solvent contact at –12°C regardless of the ethanol concentration. The data suggested that 1) the conditions applied in cold‐ethanol enrichment of protein from wheat will generally preserve vital wheat gluten functionality, 2) functionality losses can be recovered by returning the solubilized fractions, and 3) the flour to which the gluten is added may require more mixing.     

不同施氮量对弱筋小麦产量与品质的影响

通过对3个弱筋小麦品种的5个不同施氮量的研究,探索出弱筋小麦对氮肥量的需求。研究结果表明,弱筋小麦产量与施氮量呈正相关关系,但品质与施氮量呈负相关关系。综合分析,在磷、钾肥确定的条件下,施纯氮量120kg hm-2的处理,既兼顾了产量又确保了弱筋粉小麦的优良品质。     

Effect of Processing on Functional Properties of Wheat Gluten Prepared by Cold‐Ethanol Displacement of Starch

Functional properties of gluten prepared from wheat flour are altered by separation and drying. Gluten was separated and concentrated by batterlike laboratory methods: development with water, dispersion of the batter with the displacing fluid, and screening to collect the gluten. Two displacing fluids were applied, water or cold ethanol (70% vol or greater, ‐13°C). Both the water‐displaced gluten (W‐gluten) and ethanol‐displaced‐ gluten (CE‐gluten) were freeze‐dried at ‐20°C as a reference. Samples were dried at temperatures up to 100°C using a laboratory, fluidized‐bed drier. Tests of functionality included 1) mixing in a mixograph, 2) mixing in a farinograph, and 3) the baked gluten ball test. Dough‐mixing functionality was assessed for Moro flour (9.2% protein) that was fortified up to 16% total protein with dried gluten. In the mixograph, CE‐gluten (70°C) produced improved dough performance but W‐gluten (70°C) degraded dough performance in proportion to the amount added in fortification. In the microfarinograph, there was a desirable and protein‐proportional increase in stability time for CE‐gluten (70°C) but no effect on stability for W‐gluten (70°C). Baking was evaluated using the baked gluten ball test and the percentage increase in the baked ball volume relative to the unbaked gluten volume (PIBV). PIBV values were as high as 1,310% for freeze‐dried CE‐gluten and as low as 620% for W‐gluten dried at 70°C. PIBV for CE‐gluten was reduced to 77% of the freeze‐dried control by fluid‐bed drying at 70°C. Exposure of CE‐gluten to 100°C air gave a PIBV that was 59% of the reference, but this expansion was still greater than that of W‐gluten dried at 70°C. The highest values of PIBV occurred at the same mixing times as the peak mixograph resistance.     

Mixograph Responses of Gluten and Gluten‐Fortified Flour for Gluten Produced by Cold‐Ethanol or Water Displacement of Starch from Wheat Flour

The total protein of gluten obtained by the cold‐ethanol displacement of starch from developed wheat flour dough matches that made by water displacement, but functional properties revealed by mixing are altered. This report characterizes mixing properties in a 10‐g mixograph for cold‐ethanol‐processed wheat gluten concentrates (CE‐gluten) and those for the water‐process concentrates (W‐gluten). Gluten concentrates were produced at a laboratory scale using batter‐like technology: development with water as a batter, dispersion with the displacement fluid, and screening. The displacing fluid was water for W‐gluten and cold ethanol (≥70% vol, ‐12°C) for CE‐gluten. Both gluten types were freeze‐dried at ‐10°C and then milled. Mixograms were obtained for 1) straight gluten concentrates hydrated to absorptions of 123–234%, or 2) gluten blended with a low protein (9.2% protein) soft wheat flour to obtain up to 16.2% total protein. The mixograms for gluten or gluten‐fortified flour were qualitatively and quantitatively distinguishable. We found differences in the mixogram parameters that would lead to the conclusion of greater stability and strength for CE‐gluten than for W‐Gluten. Differences between the mixograms for these gluten types could be markedly exaggerated by increasing the amount of water to the 167–234% range. Mixograms for evaluation of gluten have not been previously reported in this hydration range. Mixograms for fortification suggest that less CE‐gluten than W‐gluten would be required for the same effect.     

Effects of Wheat Starch and Gluten on Tortilla Texture

Wheat starches were isolated from three wheat flours. Two vital wheat glutens, one from a commercial source and another one isolated from straight-grade flour, were combined with wheat starches to form reconstituted flours with a protein level of 10%. Several characteristics of tortillas made with the hot-press method were measured. No significant difference (P < 0.05) occurred in texture of tortillas made with hard wheat gluten and soft wheat gluten. Wheat starches did not have any significant (P < 0.05) effect on tortilla stretchability or foldability. Analysis of variance confirmed that wheat starch and gluten had limited effects on tortilla texture. The possible reasons were that the solubles of wheat flour were not included, and the shortening in the tortilla formula interfered with the interaction of gluten and starch.     

Proteins Extracted by Water or Aqueous Ethanol During Refining of Developed Wheat Dough to Vital Wheat Gluten and Crude Starch as Determined by Capillary‐Zone Electrophoresis (CZE)

Fluids applied to large‐sale, technical separation of wheat starch and protein also extract soluble proteins. The degree and rate of extraction and the specific components extracted depend on the flour, the flour hydration and development, the starch‐displacing fluid composition, the temperature, and the mechanical processing method. This study sought to identify major extracted protein groups using high‐performance capillary zone electrophoresis (CZE) applied directly to fluids obtained during laboratory‐scale technical separations. A dough‐ball or compression separation method was applied using a Glutomatic system and a batter or dispersion method was applied using a a McDuffie mixer and Pharmasep vibratory separator. Process fluids were water at 22°C to model commercial practice and 70 vol% ethanol in water at ‐13°C to model the cold ethanol process being developed here. Data were referenced to use of 70 vol% ethanol in water at 22°C in the Glutomatic compression method. The dough processed by each method was developed by mixing to a separable state. When flooded with excess water, this dough immediately released starch and water‐soluble or albumin proteins. When flooded with excess cold aqueous ethanol, neither the albumin nor gliadin proteins appeared in significant amounts until the bulk of the starch had been displaced, regardless of the mechanical method. Even with extraction and manipulation well beyond that necessary for starch displacement, the net amount of gliadin proteins dissolved was only ≈10% of that available from wet developed dough using 70 vol% ethanol at 22°C. There was more gliadin protein in the fluids at earlier stages of processing when the batter dispersion method was applied using cold ethanol. The most common soluble proteins revealed in the electrophoresis patterns for the batter compression method using cold aqueous ethanol were initially albumins and later γ‐gliadins. Albumins not appearing as soluble in cold 70 vol% ethanol were found in the insoluble crude starch, suggesting their precipitation in the dough fluids during the change from free water to cold aqueous ethanol. These results establish that some protein is dissolved during starch displacement by cold aqueous ethanol, but that the amounts may be limited by control of the mechanical working of the dough in the presence of the displacing fluids.     

Substitution of Concentrated Ethanol for Water in the Laboratory Washing Fractionation of Protein and Starch from Hydrated Wheat Flour

An unprecedented, ethanol-based, washing process was used at a laboratory scale to produce both concentrated protein and starch fractions from hydrated wheat flour. In this multistep process, flour was first hydrated and mixed to a batter and then chilled and rested. The cold batter was then mixed and washed in chilled and concentrated ethanol using a modified device that normally applies the water-based Martin process. Control of the separation was affected by each of these steps. For instance, the hydration of the flour, the time of mixing, the temperature of the wash, the ethanol concentration, and the time of washing were influential. The method produced a gluten concentrate similar in yield and protein content to that reported for a pilot-scale Martin process but without the need for added salt. Notably, ethanol washing resulted in nonsticky, partially disintegrated curds that dried easily, whereas water washing resulted in a sticky, glutinous, cohesive mass that dried slowly. The process has commercial potential to reduce water and energy use, reduce wastewater generation and environmental impact, and improve product recovery. The process also has the potential to reduce the capital complexity of the drying step and create convenient opportunities for protein subfractionation.     

Impact of Bran Addition on Water Properties and Gluten Secondary Structure in Wheat Flour Doughs Studied by Attenuated Total Reflectance Fourier Transform Infrared Spectroscopy

The aim of this work was to elucidate the underlying physical mechanism(s) by which bran influences whole grain dough properties by monitoring the state of water and gluten secondary structure in wheat flour and bran doughs containing 35–50% moisture and 0–10% added bran. The system was studied with attenuated total reflectance (ATR) FTIR spectroscopy. Comparison of the OH stretch band of water in flour dough with that in H₂O‐D₂O mixtures having the same water content revealed the formation of two distinct water populations in flour dough corresponding to IR absorption frequencies at 3,600 and 3,200 cm^–1. The band intensity at 3,200 cm^–1, which is related to water bound to the dough matrix, decreased and shifted to lower frequencies with increasing moisture content of the dough. Addition of bran to the dough caused redistribution of water in the flour and bran dough system, as evidenced by shifts in OH stretch frequency in the 3,200 cm^–1 region to higher frequencies and a reduction in monomeric water (free water). This water redistribution affected the secondary structure of gluten in the dough, as evidenced by changes in the second‐derivative ATR‐FTIR difference spectra in the amide I region. Bran addition caused an increase in β‐sheet content and a decrease in β‐turn (β‐spiral) content. However, this bran‐induced transconformational change in gluten was more significant in the 2137 flour dough than in Overley flour dough. This study revealed that when bran is added to flour dough, water redistribution among dough components promotes partial dehydration of gluten and collapse of β‐spirals into β‐sheet structures. This transconformational change may be the physical basis for the poor quality of bread containing added bran.     

Wheat Gluten Polymer Structures: The Impact of Genotype,Environment, and Processing on Their Functionality in Various Applications

For a number of applications, gluten protein polymer structures are of the highest importance in determining end‐use properties. The present article focuses on gluten protein structures in the wheat grain, genotype‐ and environment‐related changes, protein structures in various applications, and their impact on quality. Protein structures in mature wheat grain or flour are strongly related to end‐use properties, although influenced by genetic and environment interactions. Nitrogen availability during wheat development and genetically determined plant development rhythm are the most important parameters determining the gluten protein polymer structure, although temperature during plant development interacts with the impact of the mentioned parameters. Glutenin subunits are the main proteins incorporated in the gluten protein polymer in extracted wheat flour. During dough mixing, gliadins are also incorporated through disulfide‐sulfhydryl exchange reactions. Gluten protein polymer size and complexity in the mature grain and changes during dough formation are important for breadmaking quality. When using the gluten proteins to produce plastics, additional proteins are incorporated in the polymer through disulfide‐sulfhydryl exchange, sulfhydryl oxidation, β‐eliminations with lanthionine formation, and isopeptide formation. In promising materials, the protein polymer structure is changed toward β‐sheet structures of both intermolecular and extended type and a hexagonal close‐packed structure is found. Increased understanding of gluten protein polymer structures is extremely important to improve functionality and end‐use quality of wheat‐ and gluten‐based products.     

Effect of Environment and Genotype on Durum Wheat Gluten Strength and Pasta Viscoelasticity

Data on the quality of durum wheat genotypes grown under eight environments (site-year combinations) were evaluated to determine the relative effects of genotype and environment on quality characteristics associated with gluten strength, protein content, and pasta texture. The 10 durum wheat genotypes assessed in this study represented a range of gluten strength types from the very strong U.S. desert durum genotype, Durex, to the medium strength Canadian genotype, Plenty. Considerable genetic variability was detected for all quality characteristics studied. Genotype-environment interaction was significant for all quality parameters evaluated, with the exception of mixograph development time. Genotypeenvironment interaction was most important in determining protein content and least important in determining gluten index, gluten viscoelasticity, and SDS sedimentation volume. The nature of the genotype-environment interaction was evaluated by determining the number of significant crossover (rank change) interactions. There was at least one significant crossover interaction between pairs of genotypes and environments for five of eight quality traits tested. Of 45 genotype pairs, eight and six showed significant crossover interactions for protein content and pasta disk viscoelasticity, respectively. Significant crossover interactions were at least partially due to the differential response of Canadian genotypes as compared with U.S. genotypes. With the exception of protein content and pasta disk viscoelasticity, our results suggest that among the selected sample of 10 genotypes, genotype-environment interactions were minor and due primarily to changes in magnitude rather than changes in rank.     

Impact of Waxy,Partial Waxy,and Wildtype Wheat Starch Fraction Properties on Hearth Bread Characteristics

Thirteen different wheat cultivars were selected to represent GBSS mutations: three each of wildtype, axnull, and bxnull, and two each of 2xnull and waxy. Starch and A‐ and B‐granules were purified from wheat flour. Hearth bread loaves were produced from the flours using a small‐scale baking method. A‐granules purified from wildtype and partial waxy (axnull, bxnull, and 2xnull) starches have significantly higher gelatinization enthalpy and peak viscosity compared with B‐granules. A‐ and B‐granules from waxy starch do not differ in gelatinization, pasting, and gelation properties. A‐ and B‐granules from waxy starch have the highest enthalpy, peak temperature, peak viscosity, breakdown, and lowest pasting peak time and pasting temperature compared with A‐ and B‐granules from partial waxy and wildtype starch. Waxy wheat flour has much higher water absorption compared with partial waxy and wildtype flour. No significant difference in hearth bread baking performance was observed between wildype and partial waxy wheat flour. Waxy wheat flour produced hearth bread with significantly lower form ratio, weight, a more open pore structure, and a bad overall appearance. Baking with waxy, partial waxy, and wildtype wheat flour had no significant effect on loaf volume.     

氮素供应对强筋小麦产量和品质的影响

选择淮北地区两种土壤类型进行田间试验,研究氮素供应对强筋小麦的产量和品质的影响。结果表明,在一定的施氮范围内,施氮量与产量呈二次曲线关系,与品质呈直线相关关系。在对照产量为6092.15 kg hm-2的砂姜黑土,151kg hm-2的施氮量可获得最高产量7219.6 kg hm-2,蛋白质含量15.504%,湿面筋含量36.077%;174 kg hm-2的施氮量可获得最优品质,产量为7192.94 kg hm-2,蛋白质含量15.721%,湿面筋含量37.125%。对照产量为4457.4 kg hm-2的两合土,施氮量在174 kg hm-2时,可获得最优品质。经效益概算知,兼顾高产、优质、高效的施氮量是最优品质施氮量。     

氮肥运筹方法对强筋小麦产量和品质的调控效应

采用盆栽试验方法研究了氮肥运筹对强筋小麦产量和品质的影响,试验结果表明:合理运筹氮肥能够显著提高强筋小麦籽粒产量。拔节期叶面喷施氮肥显著提高了强筋小麦的株高、穗长、穗数、穗粒数和千粒重,从而促进小麦获得了最高产量;适当增加中、后期氮肥施用比例有利于改善强筋小麦品质,以拔节期和抽穗期分两次追施氮肥效果最佳。综合产量和品质效应,以拔节期喷施氮肥为最佳氮肥运筹方法。     

Demixing in Wheat Doughs—Its Influence on Dough and Gluten Rheology

Reshaping of relaxed wheat doughs leads to an increase in firmness that significantly changes the results of rheological measurements involving large uniaxial deformations of the dough, whereas the gluten properties remain unaffected. Microscopic investigations reveal that directly after kneading, starch and gluten are thoroughly mixed. However, the shaping procedure of a relaxed dough or shear-flow during rheological measurements cause a separation of gluten and starch. The dilatant behavior of the starch granules and the capacity of gluten to aggregate account for the observed dough-hardening.     

Effect of Partial Waxy Wheat on Processing and Quality of Wheat Flour Tortillas

The processing and quality of wheat flour tortillas prepared with partial waxy and normal flour were evaluated. Control procedures and formula were utilized with water absorption varied to obtain machineable doughs. Amylose content was lower in most partial waxy compared with normal wheats. The type of wheat starch did not affect most dough properties or tortilla diameter. Tortilla height and opacity were adversely affected by the decreased amount of amylose in partial waxy wheats. Sufficient leavening reactions occurred early in baking (after 10 sec) to yield an opaque disk, but some baked tortillas lost opacity and become partially transparent after baking. Starch gelatinizes, disperses, and retrogrades concurrently with the leavening reaction during the short (<30 sec) baking time. Amylose functionality during baking and cooling appears to be involved in the retention of air bubbles in tortillas.     

Effects of Temperature on Sorption of Water by Wheat Gluten Determined Using Deuterium Nuclear Magnetic Resonance

The effects of lipids and residual starch components of wheat flour gluten on gluten hydration properties were investigated using nuclear magnetic resonance (NMR) and Fourier transform infrared (FTIR) techniques. Whole or native, lipid-free, starch-free, and lipid- and starch-free gluten samples were prepared from wheat (Triticum aestivum) cv. Mercia. ²H NMR relaxation on gluten samples hydrated with deuterium oxide (D₂O) was measured over a 278–363 K temperature range. FTIR spectra were recorded in dry and fully hydrated material. Transverse relaxation (T₂) results indicated that all four gluten samples were hydrophilic in nature. There was little difference in relaxation behavior of whole and lipid-free gluten samples. T₂ values and populations of the relaxation components were very similar in each. The FTIR spectra of both samples showed an increase in extended β-sheet secondary structures on hydration. These results suggest that lipid binding in gluten, if it occurs, has little effect on wheat gluten properties. Adding starch to the gluten matrix results in an increase in water sorption on heating that may be attributed to the effects of starch gelation. However, the whole water uptake of the gluten cannot be accounted for by the contribution of the residual starch, as estimated by the effects of added starch. Extraction of residual starch required solubilization of the protein, including breaking of the disulfide bonds. This process altered the gluten structure and properties. Light microscope investigation showed that glutens with residual starch extracted were unable to form fibrillar strands on hydration. NMR and FTIR results showed greater water sorption in both samples with extracted starch than in the unextracted samples.     

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

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

Structural Modification of Gluten Proteins in Strong and Weak Wheat Dough as Affected by Mixing Temperature

The effects of temperature (≥25°C) on dough rheological properties and gluten functionality have been investigated for decades, but no study has addressed the effect of low temperature (<30°C) on gluten network attributes in flours with strong and weak dough characteristics. This study monitored changes in protein extractability in the presence and absence of reducing agents, the contents of readily accessible and SDS‐accessible thiols, and the secondary structural features of proteins in doughs from commercial hard wheat flour (HWF) and soft wheat flour (SWF) mixed at 4, 15, and 30°C. SWF mixed at 4 and 15°C showed similar mixing properties as HWF mixed at 30°C (which is the standard temperature). The effect of mixing temperature is different at the molecular level between the two flours studied. Protein features of HWF did not change as mixing temperature decreased, with the only exception being an increase in SDS‐accessible thiols. Decreasing mixing temperature for SWF caused an increase in SDS protein solubility and SDS‐accessible thiols as well as an increase in β‐turn structures at the expense of β‐sheet structures. Thus, noncovalent interactions appear to drive protein network at low temperatures (4 and 15°C), whereas covalent interactions dominate at standard mixing temperature (30°C) in doughs from both flours.