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.     

Structure of (1→3)(1→4)‐β‐d‐Glucan in Waxy and Nonwaxy Barley

The endosperm cell walls of barley are composed largely of a (1→3)(1→4)‐β‐d ‐glucan commonly known simply as β‐d ‐glucan (Wood 2001). There has been much research into the characteristics of barley β‐glucan because of the influence of this polysaccharide on performance of barley in malting and subsequent brewing of beer, and in feed value, especially for young chicks (MacGregor and Fincher 1993). The potential for β‐glucan to develop high viscosity is a problem in these uses, but from the perspective of human nutrition, this characteristic may be an advantage. The glycemic response to oat β‐glucan is inversely related to (log)viscosity (Wood et al 1994a) and there is evidence to suggest that the lowering of serum cholesterol levels associated with oat and barley products (Lupton et al 1994; Wood and Beer 1998) is at least in part due to the β‐glucan (Braaten et al 1994) and probably also its capacity to develop viscosity in the gastrointestinal tract (Haskell et al 1992).     

Effect of Specific Mechanical Energy on Protein Bodies and α-Zeins in Corn Flour Extrudates

Zeins, the storage proteins of corn, are located in spherical entities called protein bodies. The disruption of protein bodies and zein release during extrusion may influence the texture of corn-based extruded foods. In this work, chemical and microscopic studies were conducted on corn flour that had been extruded under mild to extreme conditions to determine the specific mechanical energy (SME) required to break apart protein bodies and release α-zein, and to assess changes in protein-protein interactions. Transmission electron microscopy with immunolocalization of α-zein revealed that starch granules and protein bodies remained intact under mild processing conditions (SME 35–40 kJ/kg), but under harsher conditions, protein bodies were disrupted and α-zein was released. At SME ≈100 kJ/kg, protein bodies appeared highly deformed and fused together with the α-zein released, whereas at higher SME, protein bodies were completely disrupted and α-zein was dispersed and may have formed protein fibrils. Protein in extrudates was less soluble in urea and SDS than in unprocessed corn flour, but it was readily extracted with urea, SDS, and 2-ME. This was likely due to protein aggregation upon processing due to a prevalence of hydrophobic interactions and disulfide bonds. This research directly relates SME during extrusion to chemical and structural changes in corn proteins that may affect the texture of corn-based, ready-to-eat food products.     

Low α-Amylase Starch Digestibility of Cooked Sorghum Flours and the Effect of Protein

The comparably low starch digestibility of cooked sorghum flours was studied with reference to normal maize. Four sorghum cultivars that represent different types of endosperm were used. Starch digestibilities of 4% cooked sorghum flour suspensions, measured as reducing sugars liberated following α-amylase digestion, were 15–25% lower than for cooked maize flour, but there were no differences among the cooked pure starches. After the flours were predigested with pepsin to remove some proteins, the starch digestibility of cooked sorghum flours increased 7–14%, while there was only 2% increase in normal maize; however, there was no effect of pepsin treatment on starch digestibility if the flours were first cooked and then digested. After cooking with reducing agent, 100 mM sodium metabisulfite, starch digestibility of sorghum flours increased significantly while no significant effect was observed for maize. Also, starch solubility of sorghum flours at 85 and 100°C was lower than in maize, and sodium metabisulfite increased solubility much more in sorghum than in maize. Differential scanning calorimetry results of the flour residue after α-amylase digestion did not show any peaks over a temperature range of 20–120°C, indicating that sorghum starches had all undergone gelatinization. These findings indicate that the protein in cooked sorghum flour pastes plays an important role in making a slowly digesting starch.     

Vitamin E Homologs and γ‐Oryzanol Levels in Rice (Oryza sativa L.) During Seed Development

Vitamin E homologs (tocopherols and tocotrienols) and γ‐oryzanol have gained significant attention because of their proposed health benefits and ability to increase vegetable oil stability. Changes in the levels of these phytochemicals were examined during seed development. Rapid accumulation of tocopherols, tocotrienols, and γ‐oryzanol occurred during early seed development. During the middle stage of seed development, the levels of tocopherols decreased sharply, and the levels of tocotrienols either stayed the same or decreased slightly, whereas γ‐oryzanol continued to accumulate until maturity. The levels of these compounds remained constant from maturity to 15 days postmaturity. In conclusion, rice grain for nutraceutical and functional food applications should be harvested during its immature stage to obtain the highest amount of tocopherols, whereas the amounts of the other phytochemicals that were studied can be optimized by harvesting grain at maturity.     

Mixed Linkage (1→3),(1→4)‐β‐d‐Glucans of Grasses

The mixed‐linkage (1→3),(1→4)‐β‐d ‐glucans are unique to the Poales, the taxonomic order that includes the cereal grasses. (1→3), (1→4)‐β‐Glucans are the principal molecules associated with cellulose microfibrils during cell growth, and they are enzymatically hydrolyzed to a large extent once growth has ceased. They appear again during the developmental of the endosperm cell wall and maternal tissues surrounding them. The roles of (1→3),(1→4)‐β‐glucans in cell wall architecture and in cell growth are beginning to be understood. From biochemical experiments with active synthases in isolated Golgi membranes, the biochemical features and topology of synthesis are found to more closely parallel those of cellulose than those of all other noncellulosic β‐linked polysaccharides. The genes that encode part of the (1→3),(1→4)‐β‐glucan synthases are likely to be among those of the CESA/CSL gene superfamily, but a distinct glycosyl transferase also appears to be integral in the synthetic machinery. Several genes involved in the hydrolysis of (1→3),(1→4)‐β‐glucan have been cloned and sequenced, and the pattern of expression is starting to unveil their function in mobilization of β‐glucan reserve material and in cell growth.     

α-Amylolysis of Large Barley Starch Granules

The α-amylolysis of large (volume average 16 μm) barley starch granules was studied by measuring the amount of carbohydrates solubilizing during hydrolysis, and the changes in morphology and molecular structure of the granule residues by scanning electron microscopy, particlesize analysis, size-exclusion chromatography, X-ray diffraction, and differential scanning calorimetry. X-ray diffraction showed that, in the earlier stages of α-amylolysis, both amorphous and crystalline parts of the granules were equally solubilized. More extensive hydrolysis caused a gradual decrease in A-type crystallinity and degradation of the granular structure. Scanning electron microscopy revealed that hydrolysis proceeded through pinholes, and pitted and partially hollow granule residues were formed. The lipid-complexed amylose was less susceptible to α-amylolysis than free amylose and amylopectin. Lipid-complexed amylose started leaching out of the granule residues only after half of the starch had solubilized due to the α-amylase treatment. Even though scanning electron microscopy indicated that there were intact granules left throughout the hydrolysis, the results obtained suggested that α-amylolysis of large barley starch granules proceeded rather evenly among the granules.     

Wet Processing of Barley Grains into Concentrates of Proteins, β‐Glucan,and Starch

An improved wet method was developed to process barley into fractions concentrated in protein, (1‐3)(1‐4)‐β‐d ‐glucan (BG), starch, or other carbohydrates (CHO). Alkaline concentration, solvent to barley flour ratio (SFR), and extraction temperature were evaluated for their effects on concentration and recovery of protein, BG, starch, oil, ash, and other CHO in each fraction type. Results show that the three parameters and their interactions all had significant effects, resulting in varying nutrient concentrations and recovery rates in each type of fractions. For protein fractions, protein content varied from 37.7 to 75.2%, protein recovery from 8.5 to 75.7%, and increasing alkaline concentration and SFR improved nutrient recovery. For BG fractions, BG content ranged from 21.5 to 87.0%, BG recovery from 28.6 to 78.0%, and increasing alkaline concentration decreased BG content but increased its recovery significantly. For starch fractions, starch content varied from 76.9 to 93.9%, starch recovery from 33.6 to 63.9%, and all parameters had little effect on the nutrient concentrations, but alkaline concentration and SFR improved recovery of starch, other CHO, and mass. Overall, the improved wet method was effective in concentrating the major nutrients from barley into their respective fractions, but process optimization through manipulating the three parameters is necessary to achieve a maximum concentration or recovery rate of a nutrient of interest in a specific fraction.     

β-Glucanase Activity and Molecular Weight of β-Glucans in Barley After Various Treatments

β-Glucanase activity interferes with molecular characterization of mixed-linkage (1→3)(1→4)-β-d -glucans (β-glucans). Reductions in β-glucanase activity were determined after barley cvs. Azhul, Waxbar, and Baronesse were treated with autoclaving (120°C, 45 min), calcium chloride (0.05M, 1 hr), 70% ethanol (80°C, 4 hr), hydrochloric acid (0.1N, 1 hr), oven heating (120 and 140°C, 40 min), sodium hydroxide (0.0025M, 1 hr), and 5% trichloroacetic acid (TCA) (40°C, 1 hr). High-performance size-exclusion chromatography (HPSEC) of α-amylase-treated aqueous extracts was used to demonstrate the effects of treatments on the molecular weights of β-glucans. The HPSEC system included multiple-angle, laser light scattering, refractive index, and fluorescence detectors. β-Glucanase activities, ranging from 52 to 65 U/kg of barley, were reduced by autoclaving (50–75%), hot alcohol (67–76%), oven heating (40–96%), CaCl₂ (75–95%), NaOH (76–89%), and TCA (92–96%). Some malt β-glucanase activity remained after most treatments. HCl and TCA treatments reduced extraction and molecular weights of β-glucans. Weight-average molecular weights (M_w) for β-glucans extracted with water at 23°C were low (most <8 × 10⁵). Base treatment (pH 9) and extraction at 100°C for 2.5 hr resulted in the greatest extraction of β-glucans and highest M_w. As a result, the conditions seem appropriate for measurement of physical characteristics of β-glucans in cereal products.     

Effects of Processing,Cultivar, and Environment on the Physicochemical Properties of Oat β‐Glucan

Several food regulatory agencies around the world have approved health claims for oat‐derived β‐glucan for cholesterol lowering and glycemic control. The biological efficacy of β‐glucan appears to depend both on daily intake and on physicochemical properties, such as molecular weight and viscosity. The objective of this study was to determine the effects of oat processing, genotype, and growing location on the physicochemical properties of β‐glucan. Five oat genotypes (HiFi, Leggett, CDC Dancer, Marion, and CDC Morrison) grown in two locations (Saskatoon and Kernen) were dehulled (untreated) and processed in a pilot facility through kilning (kilned, not flaked) and subsequent steaming and flaking (kilned, flaked). Untreated groats gave a relatively low Rapid Visco Analyzer (RVA) apparent viscosity (164 cP) and a low extractable β‐glucan molecular weight (332,440) but exhibited high β‐glucan solubility (90.49%). Compared with untreated groats, the kilned (not flaked) samples had significantly increased RVA apparent viscosity (314 cP) and extractable β‐glucan molecular weight (604,710). Additional processing into kilned and flaked products further increased RVA apparent viscosity (931 cP) and β‐glucan molecular weight (1,221,760), but β‐glucan solubility (63.83%) was significantly reduced. Genotype and growing environment also significantly affected β‐glucan viscosity and molecular weight, but no significant interaction effects between processing, genotype, and environment were found. Results indicate that there is potential for processors to improve the physicochemical and nutritional properties of oat end products through processing of specific oat genotypes from selected growing locations.     

Determining the Role of Starch in Flour Tortilla Staling Using α‐Amylase

Effects of α‐amylase modification on dough and tortilla properties were determined to establish the role of starch in tortilla staling and elucidate the antistaling mechanism of this enzyme. Control and amylase‐treated tortillas were prepared using a standard bake test procedure, stored at 22°C, and evaluated over four weeks. Amylase improved shelf‐stability of tortillas. The enzyme also produced a significant amount of dextrins and sugars, decreased loss of amylose solubility, and weakened starch granules. Amylopectin crystallinity increased with time, but was similar for the control and treated tortillas. Staling of tortillas appears to mainly involve the starch in the amorphous phase. As such, amylase activity does not significantly interfere with amylopectin crystallization. It is proposed that amylase partially hydrolyzed the dispersed starch (i.e., mostly amylose), starch bridging the crystalline region, and protruding amylopectin branches. Starch hydrolysis decreases the rigid structure and plasticized polymers during storage. The flexibility of tortillas results from the combined functionalities of the amylose gel and amylopectin solidifying the starch granules during storage. Protein functionality may also be involved in tortilla staling, but this needs further research.     

Microstructure of α‐Crystalline Emulsifiers and Their Influence on Air Incorporation in Cake Batter

The microstructure of α‐gel and β‐crystalline emulsifiers and their effects on cake batter foam have been studied with polarized light microscopy, confocal laser scanning microscopy (CLSM) and image analysis, freeze‐etching, and transmission electron microscopy (TEM). The emulsifiers Colco and Aroplus, which are commercial α‐gels, and the monoglyceride Dimodan P in its α‐gel and β‐crystalline forms were added to the batter in concentrations of 0.8, 2.0, 3.1, and 4.2%. Dimodan P α‐gel was also prepared with three NaCl concentrations (0.05, 0.67, and 1.35‰). The distribution of air in the foam was evaluated with density measurements and with image analysis of bubbles in optically sectioned batter. In the cake batter, all the α‐gel emulsifiers decreased the density, thereby increasing the incorporation of air, more than the β‐crystalline emulsifier, which did not have any effect on the density. There were noticeable differences in microstructure between the different α‐crystalline emulsifiers. Large, regular α‐structures seemed to increase the batter volume and interfacial area more than smaller aggregates. Adding salt in the emulsifier gel changed the structure, probably into α‐lamellar liposomes, which impaired the aerating effect at lower concentrations.     

Resistance to α‐Amylase Digestion in Four Native High‐Amylose Maize Starches

Native and processed high‐amylose maize starch (HAMS) is an important source of resistant starch (RS). The objectives of this work were to use an in vitro procedure to estimate the RS content of native granules from a series of ae‐containing HAMS genotypes, and to examine the nature of the α‐amylase resistant starch (ARS). By the method of Englyst et al (1992), RS for ae V, ae VII, ae su2, and ae du were estimated to be 66.0, 69.5, 69.5, and 40.6%, respectively. By transmission electron microscopy, most of the residual granules from ae V, ae VII, and ae su2 showed little evidence of digestion. Partially digested granules had a radial digestion pattern in the interior and an enzyme‐resistant layer near the surface. Size and chain‐length profile of constituents of ARS were similar to those of the native HAMS (unlike type 3 RS), consistent with complete hydrolysis in susceptible granule regions. Between crossed polarizers, many iodine‐stained native and residual HAMS granules had blue centers and pink exteriors, which may be due to a difference in orientation of the amylose‐iodine complexes in the exterior. Four granule color types were observed for ae du, differing in enzyme resistance. The high‐enzyme resistance of native HAMS granules may result from altered granule organization, which appears to vary among and within granules from ae‐containing genotypes.     

Effects of (1→3)(1→4)-β-d-Glucans of Wheat Flour on Breadmaking

Water-soluble nonstarch polysaccharides were extracted from commercial hard red winter wheat flour and separated into three fractions by graded ethanol precipitation. The three fractions, F15, F40, and F60, varied in polysaccharide composition. Fraction F15 was rich in watersoluble (1→3)(1→4)-β-d -glucans, and fractions F40 and F60 were rich in arabinoxylans. Addition of individual fractions to a bread formula did not affect bread loaf volume. Addition of fraction F15 to the formula improved bread crumb grain. Treatment of (1→3)(1→4)-β-D -glucan-rich fraction F15 with lichenase before its addition to the bread formula resulted in bread with poor crumb grain. Treatment of the F15 fraction with β-xylanase before its addition to the bread formula resulted in bread with slightly improved crumb grain. Presumably, the (1→3)(1→4)-β-D -glucans in fraction F15 improved crumb grain by stabilizing air cells in the bread dough and preventing coalescence of the cells. Addition of pentosan-rich fractions F40 and F60 to the bread formula did not improve crumb grain and interfered with the improving effect of (1→3)(1→4)-β-D -glucan-rich fraction F15. Hydrolysis of the arabinoxylans in flour by adding β-xylanase to the bread formula resulted in improved crumb grain.     

α‐Amylase Inhibitors from Wheat Against Development and Toxigenic Potential of Fusarium verticillioides

Fusarium verticillioides is one of the most important pathogens in maize and is a producer of fumonisin B₁ (FB₁). Although reports of its presence in wheat are scarce, the susceptibility of this cereal to fungus of the same genus motivates interest in investigating compounds present in the grain with inhibitory activity against this species. The aim of this study was to extract α‐amylase inhibitors from wheat and apply them in vitro to evaluate its effect on the development and expression of toxigenic potential of F. verticillioides. The α‐amylase inhibitors, both crude (P₀) and purified (P₁), were applied to in vitro culture containing a pathogen mycelium disc. Mycelial growth of the pathogen, glucosamine content, α‐amylase activity, and production of FB₁ were investigated. All protein extracts of wheat showed the ability to inhibit pathogen growth, especially the extract P₀ from cultivar Quartzo, which resulted in a reduction of glucosamine content (66%) and α‐amylase activity (84%). Furthermore, the protein inhibitors showed antifumonisin effect, reducing by 33 and 47% the mycotoxin production when applied as P₀ and P₁, respectively. These results suggest that α‐amylase inhibitor contributed to resistance against pathogen attack, acting in a diversified manner for each fungal species.     

Effect of Processing on Viscosity and Molecular Weight of (1,3)(1,4)‐β‐Glucan in Western Australian Oat Cultivars

Six Australian milling oat cultivars grown over two growing seasons were characterized for differences in (1,3)(1,4)‐β‐glucan (β‐glucan) viscosity, solubility, molecular weight (M_w), and the effect of processing. Oat cultivars grown in 2012 had significantly higher extracted β‐glucan viscosity from oat flour than the same oat cultivar grown in 2011 (P < 0.05, mean 137 and 165 cP, respectively). Noodle β‐glucan mean viscosity for 2012 (147 cP) was significantly higher than for 2011 (128 cP). β‐Glucan from ‘Williams’ and ‘Mitika’ oats had the highest viscosity (P < 0.05) in flour (5.92 and 5.25%, respectively) and noodles (1.64 and 1.47%, respectively) for both years, compared with the other oat cultivars. β‐Glucan (M_w) of Williams for 2012 and ‘Kojonup’ for both years were the least affected by processing, with an average drop of 33% compared with a maximum of 63% for other cultivars. Therefore, Williams showed superior β‐glucan properties to other oat cultivars studied, and can potentially provide improved health benefits. High and low β‐glucan M_w populations were found in the same elution peak after processing. Oat cultivars chosen for processing should be those with β‐glucans that are more resistant to processing, and that maintain their physiochemical properties and, therefore, bioactivity.     

Effects of Falling Number Sample Weight on Prediction of α‐Amylase Activity

Reports vary on the effects of falling number (FN) sample weight on test precision, reproducibility, and predictability of α‐amylase activity. Straight grade flours of 200 samples (25 cultivars × 2 locations × 2 N₂ levels × 2 repetitions) were assayed for α‐amylase activity and FN. Location significantly affected α‐amylase activity and FN values. The coefficients of variation (CV) for the FN tests were 5.75, 2.12, 1.93, 1.72, 4.27, and 14.47%, when assayed with sample weights of 7, 6, 5.5, 5, 4.5, and 4 g, respectively. The FN test with the greatest reproducibility between sample replicates (lowest LSD and highest ratio of range/LSD) was also produced using the 5‐g sample weight. By reducing FN sample weight from 7 to 5 g, FN values that averaged 350 sec, considered essentially sound, averaged 215 sec, thus shortening the FN test time by an average of 2 min and 15 sec when assaying sound wheat flour. The results suggest a review of the 7‐g stipulation of AACC Approved Method 56‐81B for FN in favor of reduced sample weight.     

蚕豆发芽过程中蛋白质和抗氧化能力的变化

为更好地开发利用蚕豆资源,研究蚕豆发芽过程中蛋白质及蛋白抗氧化能力的变化规律,采用凯氏定氮法测定发芽蚕豆的蛋白质含量,采用氨基酸分析仪测定氨基酸组分含量,运用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)技术观察发芽蚕豆蛋白组成的变化,通过测定发芽蚕豆蛋白的1,1-二苯基-2-三硝基苯肼(DPPH)自由基和2,2'-联氮双-(3-乙基苯并噻唑啉-6-磺酸)二铵盐(ABTS)自由基的清除能力分析其抗氧化能力。结果表明,蚕豆发芽过程中蛋白和氨基酸含量升高,促进部分蚕豆蛋白由大分子量转化分解成小分子量,提高了蚕豆蛋白的抗氧化能力,在蚕豆发芽第9天时蛋白质含量达到34.1 g·100g^-1、氨基酸含量为29.16 g·100g^-1,均达到最大值,蚕豆蛋白的自由基清除能力达到最强。综上,蚕豆的发芽过程可以增加蚕豆的营养物质成分含量,增强蚕豆的抗氧化能力。本研究结果为蚕豆蛋白、蚕豆芽等功能食品的开发利用提供了参考依据。