Starch Hydrolyzing Enzymes for Retarding the Staling of Rice Bread

Previous attempts have been made to obtain gluten‐free bread of acceptable quality for bread specific volume and crumb texture. Rice bread is a good alternative to celiac patients, but it has a very rapid staling during storage. Rice starch is more prone to retrograde during storage than wheat starch, and the special hydrophobic nature of the rice proteins requires specific enzymes to be used in the rice bread process. To retard rice bread staling, two different starch hydrolyzing enzymes (α‐amylase of intermediate thermostability and cyclodextrin glycoxyl transferase [CGTase]) have been tested and their effect on fresh bread quality and staling during storage has been evaluated. The addition of α‐amylase improved bread specific volume and crumb firmness but very sticky textures were obtained. The addition of CGTase produced even higher specific volume and similar crumb firmness with better texture. Both enzymes decreased the ability of amylopectin to retrograde during storage. The firming kinetic was lowered by the α‐amylase but not the limiting firmness, while the rice crumb from CGTase firmed quickly with a very short range of firmness increase. Results revealed that the starch hydrolysis brought about by the α‐amylase was not sufficient to retard staling. CGTase was considered a better antistaling agent because of its starch hydrolyzing and cyclizing activity.     

Effects of Pearling on Falling Number and α‐Amylase Activity of Preharvest Sprouted Spring Wheat

Preharvest sprouted wheat is often characterized by the falling number (FN) test. FN decreases in preharvest sprouted wheat as enzymatic degradation of the starchy endosperm increases. Wheat with FN values <250–275 is often discounted at the time of sale. The intent of this investigation was to evaluate the effects of debranning or pearling on the flour quality traits of five samples of wheat rated as low, med‐low, medium, med‐high, and sound that exhibited a range in FN values of 62–425 sec. Replicates of each sample were pearled for 30, 60, and 120 sec to remove portions of the outer bran layers before milling. FN was highly correlated with α‐amylase activity (r > ‐0.97) in the med‐low, medium, and med‐high FN sample sets as pearling time increased. FN increased in the medlow, medium, and med‐high FN samples by 128, 123, and 80%, respectively, after 120 sec of pearling. Pearling had no effect on flour FN of the low FN sample but α‐amylase activity was significantly decreased. Pearling had little or no effect on FN and α‐amylase activity of the sound sample. FN was moderately to strongly correlated with Rapid Visco Analyser (RVA), alveograph, and farinograph properties, and poorly correlated with protein content, flour yield, and bread loaf volume. In subsequent breadmaking studies, bread loaf volume, and crumb characteristics of flour from pearled wheat were not significantly different from loaf volume and crumb characteristics of flour from the corresponding nonpearled wheat.     

Staling of Bread: Role of Amylose and Amylopectin and Influence of Starch‐Degrading Enzymes

The present investigation aims at understanding the mechanism of bread firming during staling. Changes in the starch fraction due to the addition of amylases and their influence on the texture of bread crumb were studied during aging and after rebaking of stale bread. Pan bread was prepared by a conventional baking procedure. The influence of three different starch‐degrading enzymes, a conventional α‐amylase, a maltogenic α‐amylase, and a β‐amylase were investigated. The mechanical properties of bread were followed by uniaxial compression measurements. The microstructure was investigated by light microscopy, and starch transformations were assessed by differential scanning calorimetry (DSC) and wide‐angle X‐ray powder diffraction. Firming of bread crumb and crystallization of starch are not necessarily in agreement in systems with added amylases. Reorganization of both starch fractions, amylopectin and amylose, and the increase of starch network rigidity due to increase of polymer order are important during aging. Starch‐degrading enzymes act by decreasing the structural strength of the starch phase; for instance, by preventing the recrystallization of amylopectin or by reducing the connectivity between crystalline starch phases. On the other hand, starch‐degrading enzymes may also promote the formation of a partly crystalline amylose network and, by this, contribute to a kinetic stabilization of the starch network. Based on the results, a model for bread staling is proposed, taking into account the biphasic nature of starch and the changes in both the amylose and amylopectin fraction.     

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.     

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 Properties of Starch in Bread and Bread Model Systems: Influence of an Antistaling α‐Amylase

The influence of an antistaling α‐amylase on bread crumb and on wheat starch gels was investigated taking into account different levels of structural hierarchy. Bread was prepared by a conventional baking procedure. Starch gels were produced by heating a concentrated starch dispersion in closed molds. Bread and starch gels were characterized by compression tests, light microscopy (LM), differential scanning calorimetry, and X‐ray measurements. The α‐amylase enhanced the initial firmness of starch gels and reduced the firming rate of bread and starch gels on aging. LM revealed that amylose and amylopectin phase‐separated within the starch granules and that freshly baked control bread and starch gels showed weak birefringence which became more intense during aging. Amylase‐containing bread and starch gels exhibited strong birefringence in the amylose rich region of the granules directly after baking which did not significantly increase during aging. The enzyme hindered the retrogradation of amylopectin as detected by differential scanning calorimetry, whereas X‐ray diffraction indicated that the enzyme induced low levels of starch crystallinity which did not change during aging. It is hypothesized that the antistaling effect of the amylase is based on the capacity to partially degrade amylopectin and, by this, to hinder its recrystallization. On the other hand, the enzyme slightly degrades amylose by an endo‐mechanism which, in turn, promotes the rapid formation of a partly crystalline amylose network in fresh bread and hinders amylose rearrangements during aging.     

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 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.     

Effect of Different Enzymes on the Textural Stability of Shelf‐Stable Bread

Three enzyme systems (2 amylase‐based and 1 protease‐based) were tested in shelf‐stable bread to determine effectiveness in preserving texture during storage for eight weeks. Each enzyme was tested in formulations without glycerol or with 6% glycerol. Bread samples were analyzed to determine physical properties (crumb density, crust‐to‐crumb ratio, rate of moisture distribution from crumb to crust), mechanical properties (modulus, and a parameter [C1] describing resistance to high levels of deformation obtained by fitting stress‐strain data to a three‐parameter function), and thermal properties (thermal stability and enthalpy of transitions) as a function of storage time. Mechanical properties were further analyzed to predict asymptotic firmness. Bread firmness after storage as evaluated in terms of modulus and C1 were lower in all enzyme‐added systems, the effect of protease being the most significant. Enzymes had less effect on glycerol‐containing systems with no apparent trend. The breads had complex thermal behavior and exhibited multiple transitions. Both amylase preparations in the presence of glycerol reduced the amount of starch recrystallization.     

Factors Governing Pasting Properties of Waxy Wheat Flours

Waxy wheat (Triticum aestivum L.) contains endosperm starch lacking in amylose. To realize the full potential of waxy wheat, the pasting properties of hard waxy wheat flours as well as factors governing the pasting properties were investigated and compared with normal and partial waxy wheat flours. Starches isolated from six hard waxy wheat flours had similar pasting properties, yet their corresponding flours had very different pasting properties. The differences in pasting properties were narrowed after endogenous α‐amylase activity in waxy wheat flours was inhibited by silver nitrate. Upon treatment with protease, the extent of protein digestibility influenced the viscosity profile in waxy wheat flours. Waxy wheat starch granules swelled extensively when heated in water and exhibited a high peak viscosity, but they fragmented at high temperatures, resulting in more rapid breakdown in viscosity. The extensively swelled and fragmented waxy wheat starch granules were more susceptible to α‐amylase degradation than normal wheat starch. A combination of endogenous α‐amylase activity and protein matrix contributed to a large variation in pasting properties of waxy wheat flours.     

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.     

Gluten‐Free Bread from Sorghum: Quality Differences Among Hybrids

Gluten‐free breadmaking quality of 10 sorghum flours was compared using (relative basis) decorticated sorghum flour (70), corn starch (30), water (105), salt (1.75), sugar (1), and dried yeast (2). Batter consistency was standardized by varying water levels to achieve the same force during extrusion. Crumb properties were evaluated by digital image analysis and texture profile analysis (TPA). Significant differences (P < 0.001) in crumb grain were found among the hybrids with mean cell area ranging from 1.3 to 3.3 mm² and total number of cells ranging from 13.5 to 27.8/cm². TPA hardness values of the crumb also varied significantly (P < 0.001). Based on significant correlations (P < 0.01), starch damage, influenced by kernel hardness, was identified as a key element for these differences. Breads differed little in volume, height, bake loss, and water activity. Investigation of added ingredients on bread quality was conducted using response surface methodology (RSM) with two sorghum hybrids of opposite quality. Addition of xanthan gum (0.3–1.2% flour weight basis [fwb]) and skim milk powder (1.2–4.8% fwb) and varying water levels (100–115% fwb) were tested using a central composite design. Increasing water levels increased loaf specific volume, while increasing xanthan gum levels decreased the volume. As skim milk powder levels increased, loaf height decreased. Quality differences between the hybrids were maintained throughout the RSM.     

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.     

Understanding and Modeling the Processing‐Mechanical Property Relationship of Bread Crumb Assessed by Indentation

Measuring fundamental mechanical parameters such as Young's modulus and critical stress is a straightforward and valid approach to evaluating the physical texture of breadcrumb. The objectives of this study were to evaluate whether such fundamental mechanical properties could be measured by indentation techniques such as the AACC crumb firmness method, and then to alter breadmaking conditions so as to model the relationship between these indentation mechanical properties as a function of crumb moisture content and crumb density. Bread was baked according to a short dough process using Canadian western red spring (CWRS) wheat flour. Factors considered in the design of experiments were proofing time, water absorption, crosshead speed, and indenter diameter. Young's modulus and critical stress, measured with 12‐ and 20‐mm cylindrical indenters, were well covalidated with those obtained from a standard compression test. With increases in proofing time and water absorption, a more porous and compliant bread texture led to decreasing Young's modulus and critical stress. Our results revealed a good mapping of mechanical properties to crumb moisture content and density that were correlated to breadmaking conditions, thus permitting more precise prediction of the mechanical properties that determine bread texture.     

Effects of Commercial Hydrolytic Enzyme Additives on Canadian Short Process Bread Properties and Processing Characteristics

The effects of increasing levels of eight commercial enzymes representing four types of fungal hydrolytic enzymes (α‐amylases, proteases, xylanases, and cellulases) on Canadian short process (CSP) bread quality and processing characteristics were studied. Addition of all enzymes types at optimum levels resulted in increased loaf volume and bread score and softer crumb. All four types of enzymes appeared to be equally effective in improving bread properties compared with the controls. At high levels, greater tolerance to the addition of xylanases and cellulases compared with the addition of α‐amylases and proteases was apparent. Mixing requirements increased with increasing levels of α‐amylase but no change was apparent with the other enzymes. Addition of all enzymes reduced sheeting work requirements, indicating a dough softening effect. Optimum bread properties for all enzymes were attained within a relatively narrow range of dough sheeting work values, which presumably correspond to optimum dough handling properties. The similarity in response of bread and sheeting characteristics at optimum levels of addition for all four enzyme types suggests a common nonspecific mechanism for improver action that is probably related to water release and the resulting impact on physical dough properties.     

Effects of Additives and Storage Temperature on Staling Properties of Bagels

The effects of xanthan gum, Novamyl (a type II α‐amylase), Instant Tender‐Jel C starch (a modified starch), and GMS‐90‐SSK (a hydrated monoglyceride) on the staling properties of bagels stored at 4 and 22°C from 0–7 days were studied. Texture analysis and moisture determination were conducted on the bagels before lyophilization. Analysis of percent soluble starch, crumb pasting (Rapid Visco Analyser) and degree of amylopectin recrystallization (differential scanning calorimeter) were conducted on lyophilized bagel crumb. Novamyl‐treated bagels appeared to be the most resistant to staling over time at both storage temperatures in relation to the enthalpy of gelatinization (ΔH). Bagels containing xanthan gum, Instant Tender‐Jel C starch, and GMS‐90‐SSK showed some improvements over the control bagels, although the effects of the additives on the characteristics of the bagels varied. Bagels made with xanthan gum or monoglyceride retained slightly higher crumb moisture percentages over most days of storage. The monoglyceride‐treated bagels had higher enthalpy values, lower percentages of soluble starch, and a higher pasting profile but had the softest texture. The apparent onset of increased staling of the monoglyceride‐ treated bagels was attributed to complexes formed with the starch fractions.     

Bread Quality of Spelt Wheat and Its Starch

Flours from five spelt cultivars grown over three years were evaluated as to their breadbaking quality and isolated starch properties. The starch properties included amylose contents, gelatinization temperatures (differential scanning calorimetry), granule size distributions, and pasting properties. Milled flour showed highly variable protein content and was higher than hard winter wheat, with short dough‐mix times indicating weak gluten. High protein cultivars gave good crumb scores, some of which surpassed the HRW baking control. Loaf volume was correlated to protein and all spelt cultivars were at least 9–51% lower than the HRW control. Isolated starch properties revealed an increase in amylose in the spelt starches of 2–21% over the hard red winter wheat (HRW) control. Negative correlations were observed for the large A‐type granules to bread crumb score, amylose level, and final pasting viscosity for cultivars grown in year 1999 and to pasting temperature in 1998 samples. Positive correlations were found for the small B‐ and C‐type granules relative to crumb score, loaf volume, amylose, and RVA final pasting viscosity for cultivars grown in 1999, and to RVA pasting temperature for samples grown in 1998. The environmental impact on spelt properties seemed to have a greater effect than genetic control.     

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.     

Characteristics of Modern Triticale Quality: The Relationship Between Carbohydrate Properties, α‐Amylase Activity,and Falling Number

Triticale is a high‐yielding cereal crop with potential to increase grain production for human consumption over the coming decades. Minimal targeted selection has been conducted to produce cultivars with α‐amylase, amylose, and nonstarch polysaccharide (NSP) content appropriate for a milling market. Nevertheless, genotypic variability exists. Standard quality screening methods used for wheat, including pasting properties, falling number, and quantification of α‐amylase activity were employed to assess the environmental and genotypic variability among modern triticale cultivars and to investigate the suitability of these tests for triticale. Samples of 11 triticale lines from four environments were compared with five wheat cultivars bred for various end uses. Triticale exhibited a greater range than wheat for most tested variables, and the ranges usually overlapped. Triticale exhibited higher NSP content, generally equivalent pasting properties, higher α‐amylase activity, and lower falling number on average compared with wheat checks. However, low falling number was not indicative of high α‐amylase activity; the relationship with NSP level and other factors is discussed, and caution is recommended for interpretation of previous research. Three cultivars with equivalent α‐amylase activity to wheat and two with partially waxy starch were identified. These findings have great significance for research and the emerging triticale milling market.