Fermented Sorghum as a Functional Ingredient in Composite Breads

Whole sorghum flour was fermented (a five‐day natural lactic acid fermentation) and dried under forced draught at 60°C, and evaluated for its effect on sorghum and wheat composite bread quality. In comparison with unfermented sorghum flour, fermentation decreased the flour pH from 6.2 to 3.4, decreased total starch and water‐soluble proteins, and increased enzyme‐susceptible starch, total protein, and the in vitro protein digestibility (IVPD). Fermentation and drying did not decrease the pasting temperature of sorghum flour, but slightly increased its peak and final viscosity. In comparison with composite bread dough containing unfermented sorghum flour, fermented and dried sorghum flour decreased the pH of the dough from 5.8 to 4.9, increased bread volume by ≈4%, improved crumb structure, and slightly decreased crumb firmness. IVPD of the composite bread was also improved. Mixing wet fermented sorghum flour directly with wheat flour (sourdough‐type process) further increased loaf volume and weight and reduced crumb firmness, and simplified the breadmaking process. It appears that the low pH of fermented sorghum flour inactivated amylases and increased the viscosity of sorghum flour, thus improving the gas‐holding capacity of sorghum and wheat composite dough. Fermentation of sorghum flour, particularly in a sourdough breadmaking process, appears to have considerable potential for increasing sorghum utilization in bread.     

Impact of Wheat Fiber on Frozen Dough Shelf Life and Bread Quality

Freezing and prolonged frozen storage of dough results in constant deterioration in the overall quality of the final product. In this study the effect of wheat bran and wheat aleurone as sources of arabinoxylan (AX) on the quality of bread baked from yeasted frozen dough was investigated. Wheat fiber sources were milled to pass through a 0.5 mm screen, prehydrated for 15 min, and incorporated into refined wheat flour at 15% replacement level. Dough products were prepared from refined flour (control A), whole wheat flour (control B), aleurone composite flour (composite flour A), and bran composite flour (composite flour B) and stored at –18°C for 28 weeks. Dough samples were evaluated for breadmaking quality at zero time, 14 weeks, and 28 weeks of storage. Quality parameters evaluated were loaf weight, loaf specific volume, and crumb firmness. Composite flour bread samples showed the most resistance to freeze damage (less reduction in the overall product quality), indicating a possible role of some fiber components (e.g., AX) in minimizing water redistribution in the dough system and therefore lessening adverse modifications to the gluten structure. The data suggest that the shelf life of frozen dough and quality of obtained bread can be improved with the addition of an AX source.     

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.     

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.     

Application of Pregerminated Brown Rice for Breadmaking

Pregerminated brown rice (PGBR) prepared by immersing in water was used for breadmaking, and effects on the dough properties and bread qualities were studied to compare with the ungerminated brown rice (BR). The substitution of BR or PGBR for wheat flour produced smaller specific volume in bread than in the control bread without BR or PGBR along with the increasing amount of substitution. However, the bread samples containing BR or PGBR suppressed staling during storage. The improving effect was especially obvious for substitutions of 10 and 20% PGBR as compared with BR. PGBR made viscous dough and retarded the staleness of bread compared with BR. γ‐Aminobutyric acid and oryzanol did not contain in the final BR and PGBR substituted bread, and phytic acid was decomposed ≈54 and 45% for 30% BR and 30% PGBR substituted breads, respectively. But ferulic acid was quite stable in the final baked product. As a result, substitution of PGBR for wheat flour improved the bread quality.     

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.     

Effects of Monoglycerides of Varying Fatty Acid Chain Length and Mixtures Thereof on Sponge‐and‐Dough Breadmaking Quality

Monoglycerides are widely used in the baking industry because of their antistaling effects, mainly suppressing crumb firming. Commercial monoglycerides are normally prepared from hydrogenated fats, with stearate being the most common fatty acid. In a previous study, monoglycerides such as monopalmitate (C16) and monostearate (C18) had positive effects on Canadian short process bread but no improvements on sponge‐and‐dough process (SDP) bread. The objective of this study was to investigate the effects of saturated monoglycerides of varying fatty acid chain length (C14–C22) on SDP breadmaking quality by using volume judgment, crumb image analysis, and texture measurements. Higher levels (1.00–1.50%) of all monoglycerides (C14, C16, and C18) significantly (P < 0.05) increased loaf volume and cell diameter. The larger cell diameter with increasing levels of these monoglycerides may have resulted from softer, more extensible dough handling properties and greater gas cell stability during baking. Addition of C16 and C18 caused the largest increase in crumb softness with increasing monoglyceride levels but showed relatively low resilience, which might be related to larger loaf volume (i.e., lower density of bread). However, addition of blended monoglycerides C14+C16 increased crumb softness and loaf volume while partially retaining resilience. Each monoglyceride had a different function in breadmaking quality and somewhat positive effects on SDP.     

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.     

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.     

Effects of Mutant Thermostable α‐Amylases on Rheological Properties of Wheat Dough and Bread

Thermostable mutant α‐amylases (21B, M111, and M77) with various degrees of thermostability were purified from Bacillus amyloliquefaciens F and used as improvers for breadmaking. Test baking with the mutant enzymes was conducted using the long fermentation sponge‐dough method. Addition of an appropriate amount of mutant α‐amylases to the ingredients distinctly increased the specific volume of the bread and improved the softness of breadcrumb as compared with the addition of Novamyl (NM), an exo‐type α‐amylase. M77 was the most effective in retarding the staleness of breadcrumb. The softness of breadcrumb during storage, however, was not correlated with the thermostability. All mutant α‐amylases weakened the mixing property of the dough, whereas they strengthened the property of fermented dough. Especially, M77 and NM had different effects on the dough properties, but their bread qualities were similar to each other. The strong tolerance of M77 dough to the long baking process might be due to the production of hydrolyzed starches, oligosaccharides in the range of maltopentaose to maltohexaose, as compared with NM. Therefore, in the light of present findings, these mutant α‐amylases are possible substitutes for NM as bread improvers.     

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.     

Restoration of Breadmaking Properties to Frozen Dough by Addition of Sugar and Yeast and Subsequent Processing

We prepared bread dough A (a mixture of wheat flour, sugar, salt, and water), bread dough B (a mixture of bread dough A and yeast), and bread dough C (first‐proofed, molded, and second‐proofed bread dough B) and froze them at –20°C for six days. They were thawed at 4°C for 16 hr and subjected to their breadmaking processes. The results indicated that breadmaking properties (bread height [mm] and specific volume [cm³/g]) after bread dough A and B processes were the same as those of control bread dough (unfrozen dough). However, in the case of bread dough C, the resulting bread showed depression of the properties. The amount of centrifuged liquid from thawed bread dough C increased. Sugar was added to thawed bread dough C (bread dough C‐1), and then yeast was further added to bread dough C‐1 (bread dough C‐2), and they were subjected to the breadmaking process. The results showed that the breadmaking properties of bread dough C‐2 were the same as those of the control. It was further found that when the first proof step in the bread dough C‐2 process was omitted, the breadmaking properties were depressed. Frozen and thawed bread dough C was packed into a plastic tube, and extension of the dough was compared with that of control dough under reduced pressure. Bread dough C extended to 50 mm, compared with 70 mm for control dough. First proof, mold, and second proof steps of dough C‐2 caused it to extend to the same height as control dough. It was concluded that the increased amount of the separated liquid in thawed dough C caused depression of breadmaking properties resulting from lack of water in the appropriate places to provide the expected properties, but these properties could be restored to the levels of control bread dough by the addition of sugar and yeast following the first proof, mold, and second proof steps.     

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.     

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.     

Modeling of Dough Mixing Profile Under Thermal and Nonthermal Constraint for Evaluation of Breadmaking Quality of Hard Spring Wheat Flour

This research was initiated to investigate associations between flour breadmaking traits and mixing and empirical dough rheological properties under thermal stress. Thirty hard spring wheat flour samples were analyzed by a Mixolab standard procedure. Mixolab profiles were divided into six different stages, and torque measurements of individual stages were modeled by nonlinear curve fitting using a compound of two solution searching procedures, multidimensional unconstrained nonlinear minimization and genetic algorithm. Mixing patterns followed exponential equations. Dough torque patterns under heat constraint, specifically dough thermal weakening and pasting profiles, were described by a sigmoid logistic equation as a function of time. Dough stability during heating appeared important for bread loaf volume increase from significant correlations between bread loaf volume and parameters generated from models of a dough thermal weakening stage. Multivariate continuum regression was employed to calibrate prediction models of baking traits using Mixolab parameters. Coefficients of determination estimated from prediction models and cross‐validation were greater than 0.98 for bake water absorption, mixing time, and bread loaf volume, indicating that the Mixolab parameters have a potential to enhance evaluation of flour breadmaking quality.     

Antifreeze Activity of γ‐Polyglutamic Acid and Its Impact on Freezing Resistance of Yeast and Frozen Sweet Dough

This study investigated the antifreeze activity (AF) of γ‐polyglutamic acid (γ‐PGA), freezing resistance of yeast cells and sweet dough, and the mechanism influenced by γ‐PGA. Properties studied included AF of γ‐PGA, water‐holding capacity of flour, survival ratio and oxidation resistance capability of yeast cells, ice melting enthalpy (ΔH), and fermentation and breadmaking properties of sweet dough. The AF of γ‐PGA was 8.03 g of unfrozen water/g of sample, indicating good AF. γ‐PGA was tested on yeast cells and sweet dough stored frozen for 0, 1, 2, 4, and 8 weeks at four levels (0, 0.5, 1, and 3%). Survival ratio of yeast cells with γ‐PGA was significantly higher than the corresponding control. A possible mechanism might be related to the modulation of oxidation resistance capability of yeast cells by γ‐PGA. A decrease in glutathione release from frozen yeast cells and an increase in water‐holding capacity of wheat dough were observed with the addition of γ‐PGA. In the presence of γ‐PGA, ΔH, ice melting temperature, and proofing time of frozen sweet dough decreased significantly, and fermentation parameters improved, compared with the corresponding control sample. Specific volume of bread made from frozen sweet dough with 0.5, 1, and 3% γ‐PGA increased by 6.3, 8.9, and 3.3%, respectively, after 8 weeks of frozen storage. γ‐PGA enhanced the freezing resistance of yeast cells and sweet dough effectively, and the effect on specific volume of bread was not linear, with 1% showing better results.     

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.     

Effects of tyrosinase and laccase on oat proteins and quality parameters of gluten-free oat breads

Effects of a Trichoderma reesei tyrosinase (TYR) and a Trametes hirsuta laccase (LAC) on breadmaking performance of gluten-free oat flour were investigated by SDS-PAGE analysis of oat protein fractions, large deformation rheology, and microscopy of the doughs, as well as on the basis of specific volume and firmness of the gluten-free breads. TYR induced the formation of higher molecular weight proteins in the SDS-PAGE assay. Microscopical analysis showed more intensive protein aggregation in the TYR-treated dough than in the dough without TYR. TYR also increased the firmness of the dough, which was assumed to be because of the cross-linking of oat globulins. LAC did not affect the oat globulins. TYR alone, or together with a commercial Thermomyces lanuginosus xylanase (XYL), increased significantly the specific volume of the gluten-free oat bread. A combination of TYR and XYL also increased the softness of the bread, whereas a combination of LAC and XYL improved the specific volume but did not affect the softness of oat bread. The results thus indicate that cross-linking of oat globulins by TYR, especially with the addition of XYL, was beneficial for improving the texture of gluten-free oat bread.     

Comparison of Quality Characteristics and Breadmaking Functionality of Hard Red Winter and Hard Red Spring Wheat

Various whole‐kernel, milling, flour, dough, and breadmaking quality parameters were compared between hard red winter (HRW) and hard red spring (HRS) wheat. From the 50 quality parameters evaluated, values of only nine quality characteristics were found to be similar for both classes. These were test weight, grain moisture content, kernel size, polyphenol oxidase content, average gluten index, insoluble polymeric protein (%), free nonpolar lipids, loaf volume potential, and mixograph tolerance. Some of the quality characteristics that had significantly higher levels in HRS than in HRW wheat samples included grain protein content, grain hardness, most milling and flour quality measurements, most dough physicochemical properties, and most baking characteristics. When HRW and HRS wheat samples were grouped to be within the same wheat protein content range (11.4–15.8%), the average value of many grain and breadmaking quality characteristics were similar for both wheat classes but significant differences still existed. Values that were higher for HRW wheat flour were color b*, free polar lipids content, falling number, and farinograph tolerance. Values that were higher for HRS wheat flour were geometric mean diameter, quantity of insoluble polymeric proteins and gliadins, mixograph mix time, alveograph configuration ratio, dough weight, crumb grain score, and SDS sedimentation volume. This research showed that the grain and flour quality of HRS wheat generally exceeds that of HRW wheat whether or not samples are grouped to include a similar protein content range.     

Cooking,Roasting, and Fermentation of Chickpeas,Lentils, Peas,and Soybeans for Fortification of Leavened Bread

The effects of cooking, roasting, and fermentation on the composition and protein properties of grain legumes and the characteristics of dough and bread incorporated with legume flours were determined to identify an appropriate pretreatment. Oligosaccharide content of legumes was reduced by 76.2–96.9% by fermentation, 44.0–64.0% by roasting, and 28.4–70.1% by cooking. Cooking and roasting decreased protein solubility but improved in vitro protein digestibility. Mixograph absorption of wheat and legume flour blends increased from 50–52% for raw legumes to 68–76, 62–64, and 74–80% for cooked, roasted, and fermented ones, respectively. Bread dough with cooked or roasted legume flour was less sticky than that with raw or fermented legume flour. Loaf volume of bread baked from wheat and raw or roasted legume flour blends with or without gluten addition was consistently highest for chickpeas, less for peas and lentils, and lowest for soybeans. Roasted legume flour exhibited more appealing aroma and greater loaf volume of bread than cooked legume flour, and it appears to be the most appropriate preprocessing method for incorporation into bread.