Effect of the Coenzymes NAD(P)(H) in Straight‐Dough Breadmaking on Protein Properties and Loaf Volume

The nicotinamide adenine dinucleotide coenzymes [NAD(P)(H)] are strong redox agents naturally present in wheat flour, and are indispensable cofactors in many redox reactions. Hence, it is not inconceivable that they affect gluten cross‐linking during breadmaking. We investigated the effect of increasing concentrations of NAD(P)(H) on gluten cross‐linking, dough properties, and bread volume using two flours of different breadmaking quality. Separate addition of the four nicotinamide coenzymes did not significantly affect mixograph properties. While addition of NAD⁺ hardly affected bread volume, supplementation with NADP(H) and NADH significantly decreased loaf volumes of breads made using flour of high breadmaking quality. Wheat flour incubation with NAD(P)H under anaerobic conditions increased wheat flour thiol content, while NAD(P)⁺ increased the extractability in SDS‐containing medium of the protein of the strong breadmaking flour. Based on the results, it was hypothesized that at least three reactions, competing for NAD(P)(H), occur during breadmaking that determine the final effect on protein, dough, and loaf properties. Next to coenzyme hydrolysis, the experiments pointed to coenzyme oxidation and NAD(P)(H) dependent redox reactions affecting protein properties.     

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

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

Relationship Between the Qualitative and Quantitative Compositions of Gluten Protein Types and Technological Properties of Synthetic Hexaploid Wheat Derived from Triticum durum and Aegilops tauschii

The contribution of the diploid wheat species Aegilops tauschii (Coss.) Schmall to the technological properties of bread wheat (Triticum aestivum L.) was previously studied by the investigation of synthetic hexaploids derived from tetraploid durum wheat (T. turgidum L.) and three diploid Ae. tauschii lines. The results indicated that bread volume, gluten index, SDS‐sedimentation volume, and maximum resistance of gluten were significantly influenced by the Ae. tauschii lines. To determine the relationship between technological properties and qualitative and quantitative compositions of gluten proteins, the flours of parental and synthetic lines were extracted using a modified Osborne fractionation. Gliadin and glutenin fractions were then characterized by reversed‐phase (RP) HPLC on C₈ silica gel. The HPLC patterns revealed typical differences between synthetic and parental lines. The gliadin patterns of three synthetic lines and the glutenin patterns of two synthetic lines were more similar to that of the diploid Ae. tauschii parents involved in the hybrids. In the glutenin pattern of one synthetic line, characteristics from both Ae. tauschii and the durum wheat parents were observed. The amount of total gliadin and gliadin types of the synthetic lines was mostly intermediate between those of the durum and Ae. tauschii parents. The amounts of total glutenin and glutenin types (HMW and LMW subunits) of the synthetic lines were generally higher than those of the parental lines, and the ratio of gliadins to glutenins was significantly decreased. High positive correlations were found between the amount of total glutenins, HMW, and LMW subunits and bread volume, maximum resistance and extension area of gluten, and SDS‐sedimentation volume. The ratio of gliadins to glutenin subunits had a strong negative influence on these properties. The protein content of the flours and the amount of total gluten proteins were not correlated with any of the technological properties. Results on the relationship between biochemical characteristics and the breadmaking properties indicated that wheat prebreeding would benefit from studies on protein types and quantification in the choice of parents. In addition, the potential of the diploid Ae. tauschii for improvement of breadmaking quality should be further exploited.     

Effect of Mixing Time on Gluten Recovered by Ultracentrifugation Studied by Microscopy and Rheological Measurements

The effect of mixing time on gluten formation was studied for four commercial flour mixtures. The gluten phase was separated from dough using a nondestructive ultracentrifugation method. Small deformation dynamic rheological measurements and light and scanning electron microscopy were used. The recovered gluten was relatively pure with a small amount of starch granules embedded. The protein matrix observed by microscopy became smoother with prolonged mixing. No effect of overmixing was observed on the storage modulus (G′) of gluten for any of the flours. The amount of water in gluten increased from optimum to over‐mixing for most of the flours. Increased water content during prolonged mixing was not related to an effect on G′. The Standard flour resulted in the highest water content of gluten, which increased considerably with mixing time. The Strong flour had the lowest G′ of dough, a high G′ of gluten, and no increase in gluten water content from optimum to over‐mixing. The Durum flour did not show gluten development and breakdown similar to the other flours. The differences in gluten protein network formation during dough mixing are genetically determined and depend on the flour type.     

Effect of Puroindolines on the Breadmaking Properties of Wheat Flour

The role of lipid-binding proteins from wheat seed (puroindolines) on the breadmaking properties of wheat flour was investigated by determining the relationship between breadmaking quality and puroindoline content in samples of 32 wheat cultivars. An inverse relationship was mainly explained by the link between hardness and puroindoline contents. This link is in agreement with previous results which have shown a close structural identity between basic friabilins and puroindolines. Next, the effect of puroindolines in breadmaking was investigated by performing reconstitution experiments with two puroindoline-free hard cultivars of opposite quality (Florence Aurore and Ecrin) as indicated in the screened wheat sample. Addition of 0.1% puroindolines to these flours drastically modified both the rheological properties of doughs and the structure of the bread crumb. Puroindolines are essential to the foaming properties of dough liquor, and a close relationship was found between the fine grain crumb provided by reconstituted flours with puroindolines and the fine structure of corresponding dough liquor foams. The effect of puroindolines on bread volume was mainly related to the rheological properties of wheat doughs.     

Effect of Gaseous Acetic Acid Treatment on Breadmaking Properties of Flour

Eleven different wheat flours of varying protein content were treated with gaseous acetic acid. Each wheat flour exhibited maximum breadmaking properties (bread height and specific volume) at different levels of gaseous acetic acid treatment. There was a relationship (r = +0.7384) between the level of gaseous acetic acid treatment required for maximum breadmaking properties and the protein content of the wheat flour. Wheat flours with higher protein contents were more resistant to the decrease in pH value due to acetic acid treatment.     

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.     

Differential recovery of lupin proteins from the gluten matrix in lupin-wheat bread as revealed by mass spectrometry and two-dimensional electrophoresis

Bread made from a mixture of wheat and lupin flour possesses a number of health benefits. The addition of lupin flour to wheat flour during breadmaking has major effects on bread properties. The present study investigated the lupin and wheat flour protein interactions during the breadmaking process including dough formation and baking by using proteomics research technologies including MS/MS to identify the proteins. Results revealed that qualitatively most proteins from both lupin and wheat flour remained unchanged after baking as per electrophoretic behavior, whereas some were incorporated into the bread gluten matrix and became unextractable. Most of the lupin α-conglutins could be readily extracted from the lupin-wheat bread even at low salt and nonreducing/nondenaturing extraction conditions. In contrast, most of the β-conglutins lost extractability, suggesting that they were trapped in the bread gluten matrix. The higher thermal stability of α-conglutins compared to β-conglutins is speculated to account for this difference.     

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

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

Baking Quality of Bread Wheat Cultivars Damaged by Nysius simulans

Wheat grain may be attacked by different insect species. Among them, some Heteroptera species (e.g., Aelia spp. and Eurygaster spp.) reduce wheat breadmaking quality; others, such as Nysius simulans , commonly extract water and nutrients from soy plants. The aim of this study was to assess the effect of N. simulans infestation on breadmaking quality of different bread wheat cultivars. Twelve wheat cultivars (damaged and undamaged by N. simulans ) were studied. Infested grain percentage varied between 51 and 78%, depending on cultivar. Protein and gluten quantity and quality were significantly reduced in damaged flours, as shown by gluten index, solvent retention capacity, and SDS sedimentation index. SDS‐PAGE from water‐extractable proteins evidenced an important proteolytic activity in damaged samples. Dough rheological properties showed a reduced dough viscoelasticity in damaged samples. Microbread specific volume changed from 3.26 cm³/g for samples made with undamaged flour to 2.77 cm³/g for bread made with damaged flour. No evidence for modification in starch properties was found. The infestation by N. simulans reduced wheat breadmaking quality in all cultivars studied, as a result of proteolytic activity occurring after dough hydration. Results suggest that the presence of N. simulans should be considered as a factor affecting wheat crops, mainly those located next to soy crop areas, which is the usual host for this insect.     

Effects of Oat Grain Hydrothermal Treatments on Wheat-Oat Flour Dough Properties and Breadbaking Quality

Hydrothermal treatments, which are routine in oat processing, have profound effects on oat flour dough rheological properties. The influence of roasting and steam treatments of oat grain on dough mixing and breadbaking properties was investigated when hydrothermally treated oat flour was blended with wheat flour. Roasting of oat grain (105°C, 2 hr) resulted in oat flours that were highly detrimental to wheat flour dough mixing properties and breadbaking quality. Steaming (105°C, 20 min) or a combination of roasting and steaming of oat grain significantly improved the breadbaking potential of the oat flours. The addition of oat flours increased water absorption and mixing requirements of the wheat flour dough and also decreased bread loaf volume. However, at the 10% substitution level, steamed oat flours exhibited only a gluten dilution effect on bread loaf volume when wheat starch was used as a reference. Oat flour in the breadbaking system decreased the retrogradation rate of bread crumb starch. The results indicate that adequate hydrothermal treatments of oat grain are necessary for oat flour breadbaking applications. Steamed oat flours used at a 10% level retarded bread staling without adversely affecting the loaf volume.     

Effect of Zinc and Aluminum Ions in Breadmaking

Zinc and aluminum ions as chloride or sulfate salts at 50–500 ppm metal ion (flour basis) had no detrimental effect on fermentation of yeastleavened dough. Increased mixing times (≈10–50%) due to addition of aqueous solutions of zinc (250–500 ppm) or aluminum (150–250 ppm) ions to a bread formula was overcome by withholding salt until the final mixing stage. Breads made from commercial flours (12.5% protein) containing zinc (250–500 ppm) or aluminum (150–250 ppm) ions and no oxidant had improved loaf volume and crumb grain when compared with control bread, and no off-taste. Additionally, breads with added zinc or aluminum had better crumb grains and slower firming rates when compared with breads containing optimum l -ascorbic acid (50 ppm) or potassium bromate (20 ppm). Breads made from commercial flours (11.1% protein) and three laboratory flours (11.4–13.6% protein) containing zinc (250 ppm) or aluminum (150 ppm) ions also had improved loaf volumes and crumb grains. Zinc or aluminum ions in combination with l -ascorbic acid, but not potassium bromate, had a detrimental effect on bread quality. Scanning electron microscopy of freeze-dried bread doughs revealed that zinc and aluminum ions enhanced the film-coating property of gluten. One serving (one slice, 28 g) of bread made with 250 ppm zinc ion would provide 25% of the adult recommended dietary allowance of zinc.     

Exploring Functionality of Hard and Soft Wheat Flour Blends for Improved End‐Use Quality Prediction

Blending wheat or flour to meet end‐use requirements is a critical part of the production process to deliver consistent quality products. The functionality of commercial Canadian hard red wheat flour (HWF) and soft red wheat flour (SWF) blends with ratios of 100:0, 75:25, 50:50, 25:75, and 0:100 (HWF/SWF, w/w) was investigated with new and standard methods to discern which functional properties may be indicators of bread quality and processing performance. Rheological characteristics including farinograph water absorption behavior, dough development time (DT), stability, extensigraph extensibility, and gluten aggregation of wheat flours were significantly influenced by the proportion of HWF in blends of SWF and HWF (P < 0.05). The SWF content in the blends had negative linear relationships with the protein content, lactic acid solvent retention capacity, water absorption, and GlutoPeak peak torque. Polynomial relationships were observed for sodium dodecyl sulfate sedimentation volume, DT, stability, extensibility, resistance, GlutoPeak peak time, and bread loaf volume with the amount of SWF in blends. The results indicate that linear responses may be more closely tied to protein content, whereas polynomial responses may be more indicative of protein quality and baking performance. The GlutoPeak peak time was sensitive to the addition of HWF in the blends, showing a significant change in gluten aggregation kinetics between the 0 and 25% HWF samples. Principal component analysis (PCA) confirmed that GlutoPeak peak time was a significant factor in differentiating the 0% HWF. Protein secondary structures identified in the final baked bread were also PCA factors differentiating the 0% HWF sample. Although the 0% bread sample did not deviate from the observed polynomial trend for bread loaf volume, the differences in bread protein secondary structures may translate into differences in processing tolerance in commercial settings.     

Relationships Between Gluten Rheological Properties and Hearth Loaf Characteristics

The rheological properties of fresh gluten in small amplitude oscillation in shear (SAOS) and creep recovery after short application of stress was related to the hearth breadbaking performance of wheat flours using the multivariate statistics partial least squares (PLS) regression. The picture was completed by dough mixing and extensional properties, flour protein size distribution determined by SE‐HPLC, and high molecular weight glutenin subunit (HMW‐GS) composition. The sample set comprised 20 wheat cultivars grown at two different levels of nitrogen fertilizer in one location. Flours yielding stiffer and more elastic glutens, with higher elastic and viscous moduli (G′ and G″) and lower tan δ values in SAOS, gave doughs that were better able to retain their shape during proving and baking, resulting in breads of high form ratios. Creep recovery measurements after short application of stress showed that glutens from flours of good breadmaking quality had high relative elastic recovery. The nitrogen fertilizer level affected the protein size distribution by an increase in monomeric proteins (gliadins), which gave glutens of higher tan δ and flatter bread loaves (lower form ratio).     

Effect of Enzymatic Tempering of Wheat Kernels on Milling and Baking Performance

This study examined the effect of cell‐wall‐degrading enzymes added to temper water on wheat milling performance and flour quality. An enzyme cocktail consisting of cellulase, xylanase, and pectinase and five independent variables (enzyme concentration, incubation time, incubation temperature, tempered wheat moisture content, and tempering water pH) were manipulated in a response surface methodology (RSM) central composite design. A single pure cultivar of hard red winter wheat was tempered under defined conditions and milled on a Ross experimental laboratory mill. Some treatment combinations affected flour yield from the break rolls more than that from the reduction rolls. However, a maximum for flour yield was not found in the range of parameters studied. Though treatments did not affect the optimum water absorption for breadmaking, enzyme‐treated flours produced dough exhibiting shorter mixing times and slack and sticky textures compared with the control. Regardless of differences in mixing times, specific loaf volumes were not significantly different among treatments. Crumb firmness of bread baked with flour milled from enzyme‐treated wheat was comparable to the control after 1 day but became firmer during storage up to 5 days.     

Distribution of Redox Enzymes in Millstreams and Relationships to Chemical and Baking Properties of Flour

Millstream flours, bran, pollard, and germ fractions were prepared from two Australian and two New Zealand wheat cultivars using a pilot‐scale roller mill. The distribution of six redox enzymes in milling fractions and the relationship of the enzymes to baking parameters were investigated. Lipoxygenase (LOX), dehydroascorbate reductase (DAR), and protein disulfide isomerase (PDI) tended to be higher in the tail‐end fractions of break and reduction flour streams, but the highest levels were in the bran, pollard, and germ fractions. These enzymes had moderate to strong correlations with ash content of flour. These results indicated that a considerable amount of these enzymes in the tail‐end flour streams were likely to be derived from contamination with bran, aleurone, or germ components of grain. Peroxidase (POX) tended to be higher in the break flours, but polyphenol oxidase (PPO) and ascorbate oxidase (AOX) tended to be evenly distributed in the millstream flours. These three enzymes generally had poor correlations with ash and baking parameters. LOX and DAR had a negative correlation with the baking quality of bread made in the absence of ascorbic acid (AA) but a poor correlation with improvement of bread quality made with AA. The negative correlation probably reflects the high content of ash (hence trichomes), glutathione, and protein thiols in those fractions that have high LOX and DAR, and these high‐reducing‐power components and trichomes in flour may be the actual cause of poor quality bread. PDI generally had a poor correlation with bread quality in the absence of AA but a significant positive correlation with improvement in the quality of bread made with AA. It thus seems that the endogenous levels of these six enzymes were not a limiting factor in the breadmaking process, except for PDI, the levels of which may have positively influenced breadmaking in the presence of AA.     

Breadmaking Quality of Selected Durum Wheat Genotypes and Its Relationship with High Molecular Weight Glutenin Subunits Allelic Variation and Gluten Protein Polymeric Composition

Twenty‐seven durum wheat genotypes originating from different geographical areas, all expressing LMW‐2 at Glu‐B3, and five bread wheats were evaluated for flour mixing properties, dough physical characteristics, and baking performance. Gluten polymeric composition was studied using size‐exclusion HPLC of unreduced flour protein extracts. As a group, durum wheats had poorer baking quality than bread wheats in spite of higher protein and total polymer concentrations. Durum wheats exhibited weaker gluten characteristics, which could generally be attributed to a reduced proportion of SDS‐unextractable polymer, and produced less extensible doughs than did bread wheats. However, substantial variation in breadmaking quality attributes was observed among durum genotypes. Better baking performance was generally associated with greater dough extensibility and protein content, but not with gluten strength related parameters. Extensibility did not correlate with gluten strength or SEHPLC parameters. Genotypes expressing high molecular weight glutenin subunits (HMW‐GS) 6+8 exhibited better overall breadmaking quality compared with those expressing HMW‐GS 7+8 or 20. Whereas differences between genotypes expressing HMW‐GS 6+8 and those carrying HMW‐GS 7+8 could only be attributed to variations in extensibility, the generally inferior baking performance of the HMW‐GS 20 group relative to the HMW‐GS 6+8 group could be attributed to both weaker and less extensible gluten characteristics.     

Variations in Amino Acid and Protein Contents of Wheat During Milling and Northern‐Style Steamed Breadmaking

To determine the variations of amino acid and protein during milling and steamed breadmaking, two types of wheat cultivars belonging to soft and hard wheat types were used. The results showed that losses occurred in 17 amino acids during milling. The mean loss of threonine (18.0%) was the highest, which was followed by proline (15.5%), methionine (15.1%), and histidine (15.1%). The losses of tyrosine and lysine were the lowest (8.1 and 9.7%, respectively). Losses were also found for 17 amino acids during steamed breadmaking. The highest loss was observed in alanine (17.1%), with tyrosine (12.5%) close behind, and leucine (4.3%) exhibiting the lowest loss. The mean protein contents for whole‐wheat meal, flour, and steamed bread prepared from the test materials were 15.25, 14.27, and 14.33%, respectively. This meant that protein content decreased during milling; however, a slight increase was observed during steamed breadmaking. Amino acid scores of lysine in whole‐wheat meal, flour, and steamed bread prepared from the test materials were 45.4, 41.0, and 38.2, respectively. The general trend in the variations of protein and amino acids was similar in the two wheat cultivars tested.     

Qualitative Effect of Added Gluten on Dough Properties and Quality of Chinese Steamed Bread

Glutens isolated from 15 soft red winter (SRW) wheat flours were added into a SRW wheat flour to obtain protein levels of 9.6 and 11.3% for determination of the qualitative effect of added gluten on the dough properties and quality of northern‐style Chinese steamed bread (CSB). Sodium dodecyl sulfate sedimentation (SDSS) volume of the gluten source flour exhibited positive relationships with mixograph absorption, midline peak time (MPT), and midline peak value (MPV) of the gluten‐added flours and with surface smoothness, crumb structure, and total score of CSB prepared from the gluten‐added flours regardless of protein content. Positive correlations were also observed between SDSS volume of the gluten source flour and specific volume and stress relaxation score of CSB prepared from the gluten‐added flours of 11.3% protein. The increase in protein content from 9.6 to 11.3% by gluten addition raised mixograph absorption, MPT, and MPV but had no apparent effect on resistance breakdown, dough maximum force for extension, and extensibility, and it increased CSB specific volume and crumb structure score without affecting surface smoothness, stress relaxation, and total score. Mixograph parameters exhibited significant relationships with CSB total score, indicating that they could be effective predictors of the CSB‐making quality of flours.     

Characteristics of French Bread Baked from Wheat Flours of Reduced Starch Amylose Content

Wheat genotypes of wild type, partial waxy, and waxy starch were used to determine the influence of starch amylose content on French bread making quality of wheat flour. Starch amylose content and protein content of flours were 25.0–25.4% and 14.3–16.9% for wild type; 21.2 and 14.9% for single null partial waxy; 15.4–17.1% and 13.2–17.6% for double null partial waxy; and 1.8 and 19.3% for waxy starch, respectively. Wheat flours of double null partial waxy starch produced smaller or comparable loaf volume of bread than wheat flours of wild type and single null partial waxy starch. Waxy wheat flour, despite its high protein content, generally produced smaller volume of bread with highly porous, glutinous, and weak crumb than wheat flours of wild type and partial waxy starch. French bread baked from a flour of double null partial waxy starch using the sponge-and-dough method maintained greater crumb moisture content for 24 hr and softer crumb texture for 48 hr of storage compared with bread baked from a flour of wild type starch. In French bread baked using the straight-dough method, double null partial waxy wheat flours with protein content >14.3% exhibited comparable or greater moisture content of bread crumb during 48 hr of storage than wheat flours of wild type starch. While the crumb firmness of bread stored for 48 hr was >11.4 N in wheat flours of wild type starch, it was <10.6 N in single or double null partial waxy flours. Wheat flours of reduced starch amylose content could be desirable for production of French bread with better retained crumb moisture and softness during storage.