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.     

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.     

Production of Starch with Very Low Protein Content from Soft and Hard Wheat Flours by Jet Milling and Air Classification

Jet milling is a fluid energy impact‐milling technique generally used for the ultrafine reduction of higher value materials. The efficiency of jet milling combined with air classification appears very efficient to separate starch from other wheat flour aggregate components and to produce wheat starch with very low residual protein content. Indeed, residual protein content of the starch‐rich fraction can be reduced to <2% db with a series of successive grinding and air classification operations. Lipid and pentosan contents were also reduced in the starch‐rich fraction. Nevertheless, jet milling cannot eliminate grinding differences observed between different types of wheat. Wheat hardness continues to have an effect on milling and classification yields and on the composition of air classification fractions. To obtain starch‐rich fraction with only 2% protein content, hard wheat flour required a series of at least five grinding steps, whereas only three steps are necessary for soft wheat flour. Under these conditions, hard wheat flours give 24% mass yield with 12% starch damage compared with 39% yield and a low starch damage content (6.4%) for soft wheat flour. These results highlight new prospects for the development of cereal flours, especially soft wheat flours.     

Wheat Flour Exposed to Ethanol Yields Dough with Unexpected Properties

Exposure of wheat flour to ethanol solutions followed by slow drying of the ethanol in situ alters the subsequent transformation of the flour into dough. Several types of wheat flour were exposed to small amounts of ethanol solutions so as to be “wetted” but without the appearance of a separate liquid phase. The wet sample was then dried in air. Dough was formed from the treated flour, and its rheological parameters were assessed, including time to peak strength (mixograph and farinograph) and gluten index (glutomatic). Untreated and treated flour and the dough prepared therefrom were assayed using 1D SDS‐PAGE (reducing and unreducing conditions), capillary zone electrophoresis (CZE) applied to 70% leachates with and without sonication, and differential scanning calorimetry. Both gluten index and time to peak increased as a result of the treatment, and the increase was greater for flour or enriched vital gluten with an initially low gluten index than for flour with a relatively high initial index. Endosperm fragmentation following milling of the treated flour was improved by the treatment. Thermal transitions were at lower temperatures following treatment, indicating less structural order and reduced thermal stability. No compositional differences were evident when studied with robust analytical methods. CZE of leached samples (no sonication) revealed lower amounts of accessible or detected proteins following treatment. Conformational changes and new secondary interactions, therefore, appear to cause the effect.     

Effects of Varying Weight Ratios of Large and Small Wheat Starch Granules on Experimental Straight‐Dough Bread

One commercial bread wheat flour with medium strength (11.3% protein content, 14% mb) was fractionated into starch, gluten, and water solubles by hand‐washing. The starch fraction was separated further into large and small granules by repeated sedimentation. Large (10–40 μm diameter) and small (1–15 μm diameter) starch fractions were examined. Flour fractions were reconstituted to original levels in the flour using composites of varying weight percentages of starch granules: 0% small granules (100% large granules), 30, 60, and 100% (0% large granules). A modified straight‐dough method was used in an experimental baking test. Crumb grain and texture were significantly affected. The bread made from the reconstituted flour with 30% small granules and 70% large granules starch had the highest crumb grain score (4.0, subjective method), the highest peak fineness value (1,029), and the second‐highest elongation ratio (1.55). Inferior crumb grain scores and low fineness and elongation ratios were observed in breads made from flours with starch fractions with 100% small granules or 100% large granules. As the proportion of small granules increased in the reconstituted flour, it yielded bread with softer texture that was better maintained than the bread made from the reconstituted reference flour during storage.     

Evaluation of the Functionality of Five Different Soybean Proteins in Hot‐Press Wheat Flour Tortillas

Five different soybean protein sources were added to wheat flour to increase the protein content by 15–25%, and the resulting composite flours were optimally processed into hot‐press tortillas in a pilot plant. The rheological properties of composite flours were evaluated with the farinograph, alveograph, and other wheat quality tests. Tortilla‐making qualities of the control and soybean‐fortified flours were evaluated during dough handling, hot pressing, and baking. The resulting tortillas were tested in terms of yield, physical and chemical parameters, sensory properties, color, and objective and subjective texture. The soybean‐fortified tortillas had increased yields because of the higher dough water absorption and enhanced essential amino acid scores. Among the five different soybean proteins, the defatted soybean flour (SBM1) with the lowest fat absorption index and protein dispersibility index (PDI) and the soybean concentrate produced the best fortified tortillas. The protein meals with high PDI and relatively lower water absorption index (SBM3 and SBM4) produced sticky doughs, lower alveograph P/L values, and defective tortillas. All soybean proteins produced higher yields of tortillas with an enhanced protein quality and amount of dietary fiber.     

Frozen and Unfrozen Water Contents of Wheat Flours and Their Components

The properties of frozen and unfrozen water in two different wheat flours (hard and soft), and in their main components (gluten, starch, damaged starch, water‐soluble and water‐insoluble pentosans), were described using modulated differential scanning calorimetry (DSC). As a reference, enthalpy values of crystallization (298 J/g) and melting (335 J/g) of pure water were determined from the total heat flow curves. The separation of thermal events between the reversing and nonreversing heat flows with modulated DSC was not effective due to disturbances in the modulated temperature scan. For wheat flours and their components, linear regressions described well the changes in frozen water content calculated from enthalpies of freezing (R² = 0.970–0.982) or melting (R² = 0.783–0.996). The unfrozen water content (UFWC) calculated for the hard wheat flour (29–31%, db) was close to that calculated for the soft wheat flour (30–32%). The UFWC of wheat gluten (38–47%), starch (38–42%), damaged starch (37–40%), water‐soluble pentosans (51%), and water‐insoluble pentosans (40–44%) were higher than the corresponding values for the flours. The simple summation of the contributions of each component cannot be used to estimate the overall behavior of flours.     

Characteristics of Perennial Wheatgrass (Thinopyrum intermedium) and Refined Wheat Flour Blends: Impact on Rheological Properties

Intermediate wheatgrass (IWG) (Thinopyrum intermedium) is a perennial grass with desirable agronomic traits and positive effects on the environment. It has high fiber and protein contents, which increase the interest in using IWG for human consumption. In this study, IWG flour was blended with refined wheat at four IWG‐to‐wheat ratios (0:100, 50:50, 75:25, and 100:0). Samples were analyzed for proximate composition, microstructure features, pasting properties (Micro Visco‐Amylo‐Graph device), protein solubility, and total and accessible thiols. Gluten aggregation properties (GlutoPeak tester) and mixing profile (Farinograph‐AT device) were also evaluated. IWG flour enrichment increased the pasting temperature and decreased the peak viscosity of blended flours. IWG proteins exhibited higher solubility than wheat, with a high amount of accessible and total thiols. The GlutoPeak tester highlighted the ability of IWG proteins to aggregate and generate torque. Higher IWG flour enrichment resulted in faster gluten aggregation with lower peak torque, suggesting weakening of wheat gluten strength. Finally, the addition of IWG to refined wheat flour resulted in a decrease in dough development time and an increase in consistency, likely because of the higher levels of fiber in IWG. The 50% IWG flour enrichment represents a good compromise between nutritional improvement and maintenance of the pasting properties, protein characteristics, and gluten aggregation kinetics.     

Influence of Starch and Gluten Characteristics on Rheological Properties of Wheat Flour Gel at Small and Large Deformation

Starch and gluten were isolated from 10 wheat cultivars or lines with varied amylose content. The rheological properties of 30% wheat flour gel, starch gel, and the gel of isolated gluten mixed with common starch were determined in dynamic mechanical testing under shear deformation, creep‐recovery, and compression tests under uniaxial compression. Variation of wheat samples measured as storage shear modulus (G′), loss shear modulus (G″), and loss tangent (tan δ = G″/G′) was similar between flour and starch gels and correlated significantly between flour and starch gel. The proportion of acetic acid soluble glutenin exhibited a significant relationship with tan δ of gluten‐starch mixture gel. The small difference in amylose content strongly affected the rheological parameters of flour gels in creep‐recovery measurement. Wheat flour gel with lower amylose content showed higher creep and recovery compliance that corresponded to the trend in starch gel. Compressive force of flour gel at 50 and 95% strain correlated significantly with that of starch gel. Gel mixed with the isolated gluten from waxy wheat lines appeared to have a weaker gel structure in dynamic viscoelasticity, creep‐recovery, and compression tests. Starch properties of were primarily responsible for rheological changes in wheat flour gel.     

Surface Lipids Play a Role in the Interaction of Puroindolines with Wheat Starch and Kernel Hardness

Kernel hardness is an important quality characteristic of common wheat. In this study, we investigated the role of starch surface lipids on the interaction of puroindoline proteins and starch granules through in vitro starch–protein binding experiments and flour reconstitution. SDS‐PAGE showed that there were no puroindoline proteins on the starch granule surface when surface lipids were removed or when defatted starch was incubated with puroindoline proteins. However, the puroindoline protein bands were present when defatted starch was incubated with lipids followed by purified puroindoline proteins, which indicated that starch surface lipids play a role in the binding of puroindolines to starch granules. The hardness of flour tablets and dough sheets made from reconstituted flour, which combined defatted starch incubated with lipids and puroindolines with gluten, was lower than for the control reconstituted flour, which was made from defatted starch and gluten. The results of scanning electron microscopy also showed that starch granules were embedded in the gluten in the gluten + defatted starch + lipids + puroindolines treatment. These results confirmed that starch surface lipids are involved in the interaction of puroindolines with wheat starch and kernel hardness.     

Water‐Extractable Nonstarch Polysaccharide Distribution in Pilot Milling Analysis of Soft Winter Wheat

Commercial wheat (Triticum aestivum em. Thell) flour milling produces flour streams that differ in water absorption levels because of variability in protein concentration, starch damaged by milling, and nonstarch polysaccharides. This study characterized the distribution of water‐extractable (WE) nonstarch polysaccharides (NSP) in long‐flow pilot‐milling streams of soft wheat to model flour quality and genetic differences among cultivars. Existing reports of millstream analysis focus on hard wheat, which breaks and reduces differently from soft wheat. Seven soft winter wheat genotypes were milled on a pilot‐scale mill that yields three break flour streams, five reduction streams, and two resifted streams. Protein concentration increased linearly through the break streams. WENSP concentration was low and similar in the first two break streams, which are the largest break streams. Flour recovery decreased exponentially through the reduction streams; flour ash and water‐extractable glucose and galactose polymers increased exponentially through the reduction streams. Protein concentration and WE xylan concentration increased linearly through the reduction streams. The ratio of arabinose to xylose in WE arabinoxylan (WEAX) decreased through the reduction streams, and response varied among the genotypes. Flour ash was not predictive of stream composition among genotypes, although within genotypes, ash and other flour components were correlated when measured across streams. The second reduction flour stream was the largest contributor to straight‐grade flour WEAX because of both the size of the stream and the concentration of WEAX in the stream.     

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.     

Effect of Single‐Pass and Multipass Milling Systems on Whole Wheat Durum Flour and Whole Wheat Pasta Quality

Single‐pass and multipass milling systems were evaluated for the quality of whole wheat durum flour (WWF) and the subsequent whole wheat (WW) spaghetti they produced. The multipass system used a roller mill with two purifiers to produce semolina and bran/germ and shorts (bran fraction). The single‐pass system used an ultracentrifugal mill with two configurations (fine grind, 15,000 rpm with 250 μm mill screen aperture; and coarse grind, 12,000 rpm with 1,000 μm mill screen aperture) to direct grind durum wheat grain into WWF or to regrind the bran fraction, which was blended with semolina to produce a reconstituted WWF. Particle size, starch damage, and pasting properties were similar for direct finely ground WWF and multipass reconstituted durum flour/fine bran blend and for direct coarsely ground WWF and multipass reconstituted semolina/coarse bran blend. The semolina/fine bran blend had low starch damage and had desirable pasting properties for pasta cooking. WW spaghetti was better when made with WWF produced using the multipass than single‐pass milling system. Mechanical strength was greatest with spaghetti made from the semolina/fine bran or durum flour/fine bran blends. The semolina/fine bran and semolina/coarse bran blends made spaghetti with high cooked firmness and low cooking loss.     

Study of a Small‐Scale Laboratory Wet‐Milling Procedure for Wheat

The objectives of this research were to study the effects of slurry specific gravity, starch table slope, slurry pumping rate, and their interactions on starch recovery and purity; and to propose a small‐scale laboratory wet‐milling procedure for wheat. First‐order and second‐order response surface regression models were developed to study the effects and interactions of slurry specific gravity, starch table slope, and slurry pumping rate on starch and gluten separation for a 100‐g wheat wet‐milling procedure. The starch and starch protein content data fit the first‐order models (R² = 0.99 and 0.96) better than the second‐order models (R² = 0.98 and 0.93). Regression results from the first‐order models indicated that specific gravity, table slope, pumping rate, and their interactions all had a significant effect on starch yield and purity. However, these effects could be simplified as the effect of the resident time of starch and gluten slurry on the starch table and the specific gravity. Starch yield increased as resident time increased and specific gravity decreased. Protein content in starch decreased as the resident time decreased and the specific gravity increased. The separation condition with specific gravity of 3 Bé, table slope of 1.04 cm/m, and pumping rate of 50 mL/min was recommended. Under this condition, starch recovery was 85.6% and protein content of starch was 0.42%, which was similar to the 1.5‐kg laboratory methods in starch recovery. Total solids recovery was 98.1%, which is similar to that from 1.5‐kg laboratory methods. These results indicated that precision of the 100‐g wheat wet‐milling procedure was similar to that of the 1.5‐kg laboratory methods.     

Effects of Transglutaminase Enzyme on Fundamental Rheological Properties of Sound and Bug‐Damaged Wheat Flour Doughs

Transglutaminase (TG) catalyzes the formation of nondisulfide covalent crosslinks between peptide‐bound glutaminyl residues and ∊‐amino groups of lysine residues in proteins. Crosslinks among wheat gluten proteins by TG are of particular interest because of their high glutamine content. Depolymerization of wheat gluten proteins by proteolytic enzymes associated with bug damage causes rapid deterioration of dough properties and bread quality. The aim of the present study was to investigate the possibility of using TG to regain gluten strength adversely affected by wheat bug proteases. A heavily bug‐damaged (Eurygaster spp.) wheat flour was blended with sound cv. Augusta or cv. Sharpshooter flours. Dynamic rheological measurements, involving a frequency sweep at a fixed shear stress, were performed after 0, 30, and 60 min of incubation on doughs made from sound or blended flour samples. The complex moduli (G* values) of Augusta and Sharpshooter doughs blended with 10% bug‐damaged flour decreased significantly after 30 min of incubation. These dough samples were extremely soft and sticky and impossible to handle for testing purposes after 60 min of incubation. To test the possibility of using TG to counteract the hydrolyzing effect of bug proteases on gluten proteins, TG was added to the flour blends. The G* values of TG‐treated sound Augusta or Sharpshooter doughs increased significantly after 60 min of incubation. The G* values of the Augusta or Sharpshooter doughs blended with bug‐damaged flour increased significantly rather than decreased after 30 and 60 min of incubation when TG was included in the dough formulation. This indicates that the TG enzyme substantially rebuilds structure of dough hydrolyzed by wheat bug protease enzymes.     

Influence of Added Starch on Mixing of Dough Made with Three Wheat Flours Differing in High Molecular Weight Subunit Composition: Rheological Behavior

The effect of mixing time (6 and 20 min) and starch content were studied on doughs prepared with three wheat flours differing in high molecular weight subunit composition. Rheological measurements were performed in dynamic oscillation: frequency and strain sweeps, stress relaxation, and in large deformation viscosity measurements. The flours were diluted with starch to cover flour protein contents of 10–15%. Water was added to keep the starch‐water ratio constant when doughs were prepared with different protein contents. By increasing the starch content of the doughs, the rheological properties approached those of a starch‐water mixture prepared with the same starch‐water ratio as in the dough. The effect of the starch granules was reinforced by prolonged mixing. This may explain the higher values of the storage modulus and relaxation times observed after 20 min of mixing. Qualities related to gluten properties, appeared more clearly in large deformation viscosity measurements.     

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.     

Physical Characteristics of Genetically Altered Wheat Related to Technological Protein Separation

Wheat protein is a technologically challenging substrate for food and nonfood applications because of its compositional diversity and susceptibility to denaturation. Genetic modification could be used to create cultivars capable of producing more uniform or focused and novel protein compositions targeted to nonfood uses. These lines could serve as expression systems for specific high‐molecular‐weight (HMW) protein polymers and would be new crops leading to more diverse agricultural opportunities. However, fundamental changes to the molecular architecture in such wheat seeds could also result in separation and processing issues, such that conventional methods of protein enrichment may need modification or even reinvention. Enriched gluten protein fractions were prepared from Bobwhite lines modified to overproduce HMW glutenin subunits Dx5 and/or Dy10. These lines serve as experimental models to test various approaches that may be taken for protein polymer enrichment. However, conventional wheat gluten enrichment based on the glutomatic as a small model of industrial methods was incapable of producing enrichment for any of the tested meal or flour, including that from the non‐transformed parent Bobwhite. Mixing in the mixograph or farinograph failed to produce standard patterns for whole kernel meal and straight‐run flour, and the normal cohesiveness of dough expected from these devices was not observed. Microscopy of stained dough samples revealed severely limited formation of normal protein networks, a capability crucial to conventional separation technology. Particle size analysis of whole kernel meal revealed a higher resistance to milling for the altered lines. Higher drying rates, lower farinograph moisture absorption, and increased thermal transition temperatures were observed. These data suggested that the native architecture of these new forms was more tightly constructed with reduced capacity for alteration by hydration and input of mechanical energy. An alternative enrichment method featuring solvation in SDS and precipitation in acetone produced coagulated (Bobwhite) or partially coagulated protein (transgenic lines producing Dx5 or Dy10) enriched to 78–85% protein with high yield.     

Starch Retrogradation and Firming of Bread Containing Hydroxypropylated,Acetylated, and Phosphorylated Cross‐Linked Tapioca Starches for Wheat Flour

The present investigation aims at understanding the role of chemically modified starch on the firmness of fresh or stale bread. Bread was prepared from wheat flour or substituted wheat flour that contained 18% chemically modified tapioca starch and 2% vital gluten. Hydroxypropylated tapioca starch (HTS), acetylated tapioca starch (ATS), phosphorylated cross‐linked tapioca starch (PTS), and native tapioca starch (NTS) were tested. Bread prepared from the substituted flour with PTS showed a firmer texture on the day of baking compared with bread prepared from NTS, HTS, and ATS. PTS retained its granular structure in the gluten network after baking and seemed to play the role of filler particles in the gluten matrix, thereby increasing firmness of fresh bread crumb. Bread prepared from the substituted flour with HTS or ATS firmed at a lower rate and showed a lower endothermic melting enthalpy of amylopectin after three days of storage compared with NTS or PTS. These findings suggest that the staling of bread containing chemically modified tapioca starch involves recrystallization of amylopectin.