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.     

Determination of secondary structural changes in gluten proteins during mixing using Fourier transform horizontal attenuated total reflectance spectroscopy

Fourier transform horizontal attenuated total reflectance (FT-HATR) was used to examine changes in the secondary structure of gluten proteins in a flour-water dough system during mixing. Midinfrared spectra of mixed dough revealed changes in four bands in the amide III region associated with secondary structure in proteins: 1317 (alpha-helix), 1285 (beta-turn), 1265 (random coil), and 1242 cm (-1) (beta-sheet). The largest band, which also showed the greatest change in second derivative band area (SDBA) during mixing, was located at 1242 cm (-1). The bands at 1317 and 1285 cm (-1) also showed an increase in SDBA over time. Conversely, the band at 1265 cm (-1) showed a corresponding decrease over time as the doughs were mixed. All bands reached an optimum corresponding to the minimum mobility of the dough as determined by the mixograph. Increases in alpha-helix, beta-turn, and beta-sheet secondary structures during mixing suggest that the dough proteins assume a more ordered conformation. These results demonstrate that it is possible, using infrared spectroscopic techniques, to relate the rheological behavior of developing dough in a mixograph directly to changes in the structure of the gluten protein system.     

Effect of Transglutaminase,Citrate Buffer,and Temperature on a Soft Wheat Flour Dough System

Transglutaminase (TGase) can improve the functional characteristics of proteins by introducing covalent bonds inter‐ or intrachains. Temperature and pH interfere with the protein structure and the catalytic activity of enzymes. Because these three factors can act synergistically, TGase, citrate buffer, and temperature were evaluated for their effects on the rheological and chemical changes in low‐protein wheat flour dough. Dough strength, measured by microextension test, significantly increased with increasing levels of TGase (8 U/g of protein), with changes in pH of the citrate buffer (pH 6.5), and by the effect of interaction between these factors. The same trend was observed in the size‐exclusion HPLC measurements, indicating that these two parameters have the effect of increasing gluten protein aggregation. Temperature had a significant effect on dough extension, measured by microextension test. The changes in secondary structure of gluten protein were investigated by FTIR second‐derivative spectra (amide I region, 1,600–1,700 cm⁻¹) and showed an increase in β‐sheet structures initiated by TGase, citrate buffer pH, and their interaction.     

Dough Properties and Bread Quality of Wheat–Barley Composite Flour as Affected by β‐Glucanase

Barley is rich in nutritionally positive compounds, but the quality of bread made of wheat–barley composite flours is impaired when a high percentage of barley is used in the mixture. A number of enzymes have been reported to be useful additives in breadmaking. However, the effect of β‐glucanase on breadmaking has scarcely been investigated. In this paper, the influence of different levels (0.02, 0.04, 0.06, and 0.08%, based on composite flour) of β‐glucanase (100,000 U/g) on the properties of dough and bread from 70% wheat, 30% barley composite flour were studied. Although dough development time, dough stability, and protein weakening value decreased after β‐glucanase addition, dough properties such as softness and elasticity as well as bread microstructure were improved compared with the control dough. β‐Glucanase also significantly improved the volume, texture, and shelf life of wheat–barley composite breads. The use of an optimal enzyme concentration (0.04%) increased specific volume (57.5%) and springiness (21%), and it reduced crumb firmness (74%) and staling rate. Bread with added β‐glucanase had a better taste, softness, and overall acceptability of sensory characteristics compared with the control bread. Moreover, the quality of wheat–barley composite bread after addition of 0.04% β‐glucanase was nearly equal to the quality of pure wheat bread. These results indicate that dough rheological characteristics and bread quality of wheat–barley composite flour can be improved by adding a distinct level of β‐glucanase.     

Changes in Distribution of Isoflavones and β‐Glucosidase Activity During Soy Bread Proofing and Baking

Bread made partially with soy may represent a viable alternative for increasing soy consumption in populations consuming Western diets. The potential health‐promoting activity of soy isoflavones may depend on their abundance and chemical form. The objective of this study was to characterize the changes in isoflavone distribution and β‐glucosidase activity during the soy breadmaking process. Soy bread ingredients were combined and mixed to form a dough that was subsequently proofed at 48°C for 1–4 hr and baked at 165°C for 50 min to produce breads. The isoflavone composition and β‐glucosidase activity in bread ingredients, doughs, and breads were monitored. Soy ingredients and wheat flour (not bread yeast) were the major contributors of the β‐glucosidase activity in bread. No degradation of isoflavones was observed during breadmaking but the isoflavone distribution was largely altered. Proofing and baking have important but different roles in changing the isoflavone distribution. Proofing converted isoflavone β‐glucosides to aglycones by highly specific β‐glucosidase activity. Thermal treatment during baking significantly decreased the isoflavone malonylglucosides and increased isoflavone β‐glucosides. Enzyme activity during proofing and the balance between formation and deconjugation of isoflavones during baking determine the isoflavone content and composition in the final product.     

Centrifuged Liquid and Breadmaking Properties of Frozen‐and‐Thawed Bread Dough

Breadmaking properties (bread height, mm, and specific volume, cm³/g ) showed marked deterioration when bread dough was frozen and stored at ‐20°C for one day. However, these properties of bread dough baked after storage for three to six days were not further deteriorated as compared with that baked after one day of storage. A large amount of liquid was oozed from the frozen‐and‐thawed bread dough. The liquid was separated from the bread dough by centrifugation (38,900 × g for 120 min at 4°C), and collected by tilting the centrifuge tube at an angle of 45° for 30 min. There was a strong correlation between the amount of centrifuged liquid and breadmaking properties (bread height and specific volume). The mechanism responsible for the oozing of liquid in frozen‐and‐ thawed bread dough was studied. The presence of yeast and salt in bread dough was suggested to be closely related to the amount of centrifuged liquid, and fermented products particularly had a large effect on the amount of centrifuged liquid.     

Viscosity and Solubility of β‐Glucan Extracted Under In Vitro Conditions from Barley β‐Glucan‐Fortified Bread and Evaluation of Loaf Characteristics

Fortifying bread with β‐glucan has been shown to reduce bread quality and the associated health benefits of barley β‐glucan. Fortification of bread using β‐glucan concentrates that are less soluble during bread preparation steps has not been investigated. The effects of β‐glucan concentration and gluten addition on the physicochemical properties of bread and β‐glucan solubility and viscosity were investigated using a less soluble β‐glucan concentrate, as were the effects of baking temperature and prior β‐glucan solubilization. Fortification of bread with β‐glucan decreased loaf volume and height (P ≤ 0.05) and increased firmness (P ≤ 0.05). Gluten addition to bread at the highest β‐glucan level increased height and volume (P ≤ 0.05) to values exceeding those for the control and decreased firmness (P ≤ 0.05). β‐Glucan addition increased (P ≤ 0.05) extract viscosity, as did gluten addition to the bread with the highest β‐glucan level. Baking at low temperature decreased (P ≤ 0.05) β‐glucan viscosity and solubility, as did solubilizing it prior to dough formulation. Utilization of β‐glucan that is less soluble during bread preparation may hold the key to effectively fortifying bread with β‐glucan without compromising its health benefits, although more research is required.     

Bran Characteristics and Bread‐Baking Quality of Whole Grain Wheat Flour

Variations in physical and compositional bran characteristics among different sources and classes of wheat and their association with bread‐baking quality of whole grain wheat flour (WWF) were investigated with bran obtained from Quadrumat milling of 12 U.S. wheat varieties and Bühler milling of six Korean wheat varieties. Bran was characterized for composition including protein, fat, ash, dietary fiber, phenolics, and phytate. U.S. soft and club wheat brans were lower in insoluble dietary fiber (IDF) and phytate content (40.7–44.7% and 10.3–17.1 mg of phytate/g of bran, respectively) compared with U.S. hard wheat bran (46.0–51.3% and 16.5–22.2 mg of phytate/g of bran, respectively). Bran of various wheat varieties was blended with a hard red spring wheat flour at a ratio of 1:4 to prepare WWFs for determination of dough properties and bread‐baking quality. WWFs with U.S. hard wheat bran generally exhibited higher dough water absorption and longer dough mixing time, and they produced smaller loaf volume of bread than WWFs of U.S. soft and club wheat bran. WWFs of two U.S. hard wheat varieties (ID3735 and Scarlet) produced much smaller loaves of bread (<573 mL) than those of other U.S. hard wheat varieties (>625 mL). IDF content, phytate content, and water retention capacity of bran exhibited significant relationships with loaf volume of WWF bread, whereas no relationship was observed between protein content of bran and loaf volume of bread. It appears that U.S. soft and club wheat bran, probably owing to relatively low IDF and phytate contents, has smaller negative effects on mixing properties of WWF dough and loaf volume of bread than U.S. hard wheat bran.     

Studies on proofing of yeasted bread dough using near- and mid-infrared spectroscopy

Dough proofing is the resting period after mixing during which fermentation commences. Optimum dough proofing is important for production of high quality bread. Near- and mid-infrared spectroscopies have been used with some success to investigate macromolecular changes during dough mixing. In this work, both techniques were applied to a preliminary study of flour doughs during proofing. Spectra were collected contemporaneously by NIR (750-1100 nm) and MIR (4000-600 cm(-1)) instruments using a fiberoptic surface interactance probe and horizontal ATR cell, respectively. Studies were performed on flours of differing baking quality; these included strong baker's flour, retail flour, and gluten-free flour. Following principal component analysis, changes in the recorded spectral signals could be followed over time. It is apparent from the results that both vibrational spectroscopic techniques can identify changes in flour doughs during proofing and that it is possible to suggest which macromolecular species are involved.     

Factors Associated with Dough Stickiness as Sensed by Attenuated Total Reflectance Infrared Spectroscopy

Attenuated total reflectance (ATR) and Fourier transform infrared (FTIR) spectroscopy have been applied in the characterization of sticky dough surfaces. The characterization provides insight in the chemical distribution of gluten protein, starch, water, and fat during dough kneading. ATR is especially useful for selective sampling of dough surfaces because the depth of penetration of radiation is quite shallow. For dough, it is calculated to be in the order of 0.5–4 μm in the mid‐infrared, ideal for measurements of stickiness effects, where only the dough surface is of interest. To investigate the cohesive and adhesive properties of the individual dough constituents, dough was peeled from the ATR plate to study the material that adhered to it. The infrared spectra obtained indicate that fat and gluten protein appear to be located at the outer sticky dough surfaces, rather than water and starch. In comparison with gluten, the fatty component showed relatively strong adhesive forces to the ATR plate; a high residual fraction was measured after peeling the dough. Gluten proteins display different cohesion and adhesion properties that are strongly dependent on their hydration state. This indicates that the degree of hydration of gluten proteins contributes to the sticky properties of (overkneaded) dough. When analyzing gluten protein in D₂O instead of a dough matrix, more or less similar results were obtained. Significant differences in amide I and amide II intensities were measured for kneaded and stretched gluten protein in comparison to untreated, wet gluten. Besides changes in the vibrational properties of the amide groups, conformational changes in the tertiary protein structure also were observed. It appears that kneading and stretching of dough results in a major decrease in α‐helices content, accompanied by an increase of extended β‐sheet conformations.     

Changes in Phytate Content in Whole Meal Wheat Dough and Bread Fermented with Phytase‐Active Yeasts

The degradation of inositol hexakisphosphate (IP6) was evaluated in whole meal wheat dough fermented with baker's yeast without phytase activity, different strains of Saccharomyces cerevisiae (L1.12 or L6.06), or Pichia kudriavzevii with extracellular phytase activity to see if the degradation of IP6 in whole meal dough and the corresponding bread could be increased by fermentation with phytase‐active yeasts. The IP6 degradation was measured after the dough was mixed for 19 min, after the completion of fermentation, and in bread after baking. Around 60–70% of the initial value of IP6 in the flour (10.02 mg/g) was reduced in the dough already after mixing, and additionally 10–20% was reduced after fermentation. The highest degradation of IP6 was seen in dough fermented with the phytase‐active yeast strains S. cerevisiae L1.12 and P. kudriavzevii L3.04. Activity of wheat phytase in whole meal wheat dough seems to be the primary source of phytate degradation, and the degradation is considerably higher in this study with a mixing time of 19 min compared with earlier studies. The additional degradation of IP6 by phytase‐active yeasts was not related to their extracellular phytase activities, suggesting that phytases from the yeasts are inhibited differently. Therefore, the highest degradation of IP6 and expected highest mineral bioavailability in whole meal wheat bread can be achieved by use of a phytase‐active yeast strain with less inhibition. The strain S. cerevisiae L1.12 is suitable for this because it was the most effective yeast strain in reducing the amount of IP6 in dough during a short fermentation time.     

Application of Polished‐Graded Wheat Grains for Sourdough Bread

Flours obtained by a specific polishing process were used to prepare sourdough and bread. Three fractions designated C‐1 (100–90%), C‐5 (60–50%), and C‐8 (30–0%) were studied. The pH, total titratable acidity levels, and buffering capacity of sourdoughs made from polished flours were significantly different from those of the control sourdough with No. 1 Canada Western Red Spring (CW), and they provided sourdough breads with better qualities than that of CW. The growth of lactic acid bacteria and yeast in polished flour sourdoughs were significantly accelerated during fermentation over that in CW sourdough. Higher maturation of polished flour sourdoughs softened the hardness of mixed dough. The intricate network of honeycomb structure gluten and uneven surface of starch granules were distinctly observed in SEM images. Substitutions of C‐5 or C‐8 sourdoughs for CW significantly increased the loaf volume and softened breadcrumbs more than CW sourdough. Flour qualities of polished flours such as suitable acidity and good buffering capacity caused by the bran fraction were effective for better growth and longer life of yeast in the dough during fermentation. Therefore, application of polished flours in sourdough bread would improve rheological properties of dough and bread as compared with CW sourdough.     

Evaluation of Various Baking Methods for Polished Wheat Flours

Flour qualities of polished wheat flours of three fractions, C‐1 (100–90%), C‐5 (60–50%), and C‐8 (30–0%), obtained from hard‐type wheat grain were used for the evaluation of four kinds of baking methods: optimized straight (OSM), long fermentation (LFM), sponge‐dough (SDM) and no‐time (NTM) methods. The dough stability of C‐5 in farinograph mixing was excellent and the maturity of polished flour doughs during storage in extensigraph was more improved than those of the commercial wheat flour (CW). There were no significant differences in the viscoelastic properties of CW dough after mixing, regardless of the baking method, while those of polished flour doughs were changed by the baking method; this tendency became clear after fermentation. The polished flours could make a better gluten structure in the dough samples after mixing or fermentation using LFM and SDM, as compared with other baking methods. Baking qualities such as specific volume and storage properties of breads from all polished flours made with SDM increased more than with other methods. In addition, viscoelastic properties of C‐5 and C‐8 doughs fermented by SDM were similar to those of CW, and the C‐5 breadcrumb showed softness similar to that of the CW. Also, SDM could make C‐5 bread with significantly higher elasticity and cohesiveness after storage for five days when compared with CW bread. Therefore, SDM with long fermentation, as compared with other baking methods, was considered suitable for use with polished flours to give better effects on dough properties during fermentation, resulting in more favorable bread qualities.     

Effects of Different Salts on Mixing and Extension Parameters on a Diverse Group of Wheat Cultivars Using 2‐g Mixograph and Extensigraph Methods

Cations of differing chaotropic capacities (LiCl, NaCl, and KCl) were used in small‐scale mixing and extensigraph studies to assess functional changes in dough behavior of wheat cultivars varying in total protein content and HMW glutenin composition. Salt addition, regardless of cationic type, caused an increase in dough strength and stability. The smaller (hydrated) and least chaotrophic cations (Li⁺<Na⁺<K⁺) effected the greatest increase in mixing time (MT) and resistance to extension (R_max) and produced the most stable resistance breakdown (RBD). The effects of different cations on mixing and extensions indicated strong intercultivar variation; differential responses to salt addition were further shown when the cultivars were grouped according to protein content and Glu‐1D or Glu‐1B genome composition. Increases in dough strength parameters due to the addition of salt were consistently more significant for cultivars showing an overexpression of Bx7 (>12% protein). In the absence of genotypic variation, a significant interactive effect of cultivar type, protein amount, and salt addition was found for all functional dough parameters except extensibility. During mixing, there was a decrease in the amount of apparent unextractable polymeric protein (%UPP) in the dough. This phenomenon was ameliorated by the presence of salt in doughs formed from weaker flours and was most pronounced early on in the mixing process (t = 100–200 sec). Results show the importance of refining 2‐g mixograph studies to include salt in the “flour and water” dough formula.     

Effects of Bran Prehydration on Functional Characteristics and Bread‐Baking Quality of Bran and Flour Blends

The effect of bran prehydration on the composition and bread‐baking quality was determined using bran and flour of two wheat varieties. Bran was hydrated in sodium acetate buffer (50mM, pH 5.3) to 50% moisture at 25 or 55°C for 1.5 or 12 h. The soluble sugar content in bran increased with prehydration. Decreases in phytate and soluble fiber were observed in prehydrated bran, but insoluble fiber was not affected by prehydration. Likewise, free phenolic content decreased, and there was little change in the content of bound phenolics in prehydrated bran. The compositional changes were greater in the bran prehydrated at 55 than at 25°C, and for 12 than for 1.5 h. Addition of prehydrated bran delayed dough development of bran and flour blends and slightly increased water absorption of dough. A higher loaf volume of fresh bread and lower crumb firmness of bread stored for 10 days were observed in bread containing bran prehydrated at 25°C than in bread containing nonhydrated bran or bran prehydrated at 55°C. The prehydration of bran at 25°C before being incorporated into refined flour for dough mixing improved bread quality by altering bran compositional properties, allowing enough water to be absorbed by fibrous materials in the bran and preventing water competition among dough constituents.     

In Situ Production of Prebiotic AXOS by Hyperthermophilic Xylanase B from Thermotoga maritima in High‐Quality Bread

In situ enrichment of bread with arabinoxylan‐oligosaccharides (AXOS) through enzymic degradation of wheat flour arabinoxylan (AX) by the hyperthermophilic xylanase B from Thermotoga maritima (rXTMB) was studied. The xylanolytic activity of rXTMB during breadmaking was essentially restricted to the baking phase. This prevented problems with dough processability and bread quality that generally are associated with thorough hydrolysis of the flour AX during dough mixing and fermentation. rXTMB action did not affect loaf volume. Bread with a dry matter AXOS content of 1.5% was obtained. Further increase in bread AXOS levels was achieved by combining rXTMB with xylanases from Pseudoalteromonas haloplanktis or Bacillus subtilis. Remarkably, such a combination synergistically increased the specific bread loaf volume. Assuming an average daily consumption of 180 g of fresh bread, the bread AXOS levels suffice to provide a substantial part of the AXOS intake leading to desired physiological effects in humans.     

Effect of Formulation and Processing Treatments on Viscosity and Solubility of Extractable Barley β‐Glucan in Bread Dough Evaluated Under In Vitro Conditions

β‐Glucan shows great potential for incorporation into bread due to its cholesterol lowering and blood glucose regulating effects, which are related to its viscosity. The effects of β‐glucan concentration, gluten addition, premixing, yeast addition, fermentation time, and inactivation of the flour enzymes on the viscosity of extractable β‐glucan following incorporation into a white bread dough were studied under physiological conditions, as well as, β‐glucan solubility in fermented and unfermented dough. β‐Glucan was extracted using an in vitro protocol designed to approximate human digestion and hot water extraction. The viscosity of extractable β‐glucan was not affected by gluten addition, the presence of yeast, or premixing. Fermentation produced lower (P ≤ 0.05) extract viscosity for the doughs with added β‐glucan, while inactivating the flour enzymes and increasing β‐glucan concentration in the absence of fermentation increased (P ≤ 0.05) viscosity. The physiological solubility of the β‐glucan concentrate (18.1%) and the β‐glucan in the unfermented dough (20.5%) were similar (P > 0.05), while fermentation substantially decreased (P ≤ 0.05) solubility to 8.7%, indicating that the reduction in viscosity due to fermentation may be highly dependent on solubility in addition to β‐glucan degradation. The results emphasize the importance of analyzing β‐glucan fortified foods under physiological conditions to identify the conditions in the dough system that decrease β‐glucan viscosity so that products with maximum functionality can be developed.     

Effects of Commercial Hydrolytic Enzyme Additives on Japanese‐Style Sponge and Dough Bread Properties and Processing Characteristics

The effects of increasing levels of eight commercial fungal enzymes enriched in four types of activity (α‐amylase, protease, xylanase, or cellulase) on Japanese‐style sponge and dough bread quality and processing characteristics have been studied using a Canadian red spring wheat straight‐grade flour. At optimum levels, the enriched α‐amylases, xylanases, and cellulases increased loaf volume and bread score and reduced crumb firmness, while the proteases only reduced crumb firmness. For α‐amylases, xylanases, and cellulases, optimum levels for crumb firmness were obtained at higher levels of addition than for loaf volume and bread score. At high levels of addition, all four enriched enzyme types reduced loaf volume and bread score and increased crumb firmness relative to optimum levels, with the proteases showing the most dramatic effects. α‐Amylases and cellulases had little impact on dough mixing requirements, while xylanases increased and proteases greatly reduced mixing requirements. All enzymes at optimum levels reduced sheeting work requirements, resulting in softer more pliable dough. Optimum bread properties for α‐amylases, xylanases, and cellulases were attained within a relatively narrow range of dough sheeting work values. This similarity in response suggests a dominant common nonspecific mechanism for their improver action, which is most likely related to water release and the resulting impact on physical dough properties.     

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

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