The combined use of hull-less barley flour and xylanase as a strategy for wheat/hull-less barley flour breads with increased arabinoxylan and (1→3,1→4)-β-D-glucan levels

Bread-making with a composite flour (CF) consisting of 60% wheat flour (WF) and 40% hull-less barley flour, increased the total and soluble (1→3,1→4)-β-D-glucan and total arabinoxylan (AX) contents of dough and bread samples, but decreased the specific bread loaf volume. A xylanase insensitive to inhibition by Triticum aestivum L. xylanase inhibitor (TAXI) and xylanase inhibiting protein (XIP), increased loaf volume by 8.8 and 20.1% for WF and CF breads, respectively. Xylanase addition not only markedly improved loaf volume of CF bread, but also increased the soluble AX content of the WF and CF dough and bread samples because of conversion of water-unextractable AX into soluble AX. The xylanase had no impact on the extractability and molecular weight of (1→3,1→4)-β-D-glucan, but (1→3,1→4)-β-D-glucan was degraded during bread-making probably because of endogenous β-glucanase activity. Taken together, the results clearly show that the combined use of hull-less barley flour and a xylanase active during bread making, lead to palatable breads with high total and soluble AX and (1→3,1→4)-β-D-glucan contents. The sum of total AX and (1→3,1→4)-β-D-glucan was 1.70% for WF bread and 3.06% for CF bread, while the sum of soluble AX and (1→3,1→4)-β-D-glucan was 0.49 and 1.41% for control WF and CF xylanase supplemented breads, respectively.     

Near infrared spectra indicate specific mutant endosperm genes and reveal a new mechanism for substituting starch with (1→3,1→4)-β-glucan in barley

Near Infrared Reflectance spectroscopy was tested as a screening method to characterise high lysine mutants from a barley collection by classification through Principal Component Analysis (PCA). Mean spectra of the samples within each cluster identified gene-specific patterns in the 2270–2360 nm region. The characteristic spectral signatures representing the lys5 locus (Risø mutants 13 and 29) were found to be associated with large changes in percentage of starch and (1→3,1→4)-β-glucan. These alleles compensated for a low level of starch (down to 30%) by a high level of (1→3,1→4)-β-glucan (up to 15–20%), thus, maintaining a constant production of polysaccharides at 50–55%, within the range of normal barley.The spectral tool was tested by an independent data set with six mutants with unknown polysaccharide composition. Spectral data from four of these were classified within the high (1→3,1→4)-β-glucan BG lys5 cluster in a PCA. Their high (1→3,1→4)-β-glucan and low starch content was verified. It is concluded that genetic diversity such as from gene regulated polysaccharide and storage protein pathways in the endosperm tissue can be discovered directly from the phenotype by chemometric classification of a spectral library, representing the digitised phenome from a barley gene bank.     

Optimization of air classification for the production of β-glucan-enriched barley flours

Barley grains contain a relevant amount of soluble fibre (i.e. β-glucans) and can be used for production of foods with healthy functions. To produce barley flours enriched in β-glucans, grain micronization and air classification of the flour was used, and a method to predict the relationship between the yield of the enriched flour and its β-glucan content was developed. This method is based on a series of sortings of micronized flour, with a progressive increase in the yield of a selected fraction; a curve for β-glucan enrichment vs. yield can then be calculated. Thus, the most suitable combination of yield and β-glucan content can be chosen. We tested this approach on two hull-less barleys with different starch type, and obtained barley flour fractions with twice the β-glucan concentration that the grain had. Enriched flour fractions with 11.2 and 15.6% β-glucans were obtained from cultivars Priora (hull-less) and CDC Alamo (waxy, hull-less), respectively, with a good flour yield (about 30%). We suggest that our method can be adopted for finely tuning the enrichment process to meet the industry's needs.     

High (1→3,1→4)-β-Glucan Barley Fractions in Bread Making and their Effects on Human Glycemic Response

Barley flour (BF) from whole naked grain and two (1→3,1→4)-β-glucan-enriched fractions, a sieved fraction (SF) and a water-extracted fraction (WF), were produced and mixed with bread wheat flour (BW), for bread-making quality evaluation. Bread was baked in a pilot plant and analysed for sensory properties, proximate composition and (1→3,1→4)-β-glucan content. Four kinds of bread were produced from different mixtures of barley ingredients and bread wheat flour: a 100% BW ((1→3,1→4)-β-glucan content: total, 0·1%; soluble, 0·1%); a 50% BW and 50% BF (2·4%; 2·0%); a 50% BW and 50% SF (4·2%; 2·8%); an 80% BW and 20% WF (6·3%; 5·7%). Eight adults were fed test meals of each of the four breads, containing the same amount (50 g) of available carbohydrate, and glycemic indices calculated from finger-prick capillary blood samples. A linear decrease in glycemic index was found for increasing (1→3,1→4)-β-glucan content. The 20% WF bread had a 28% lower glycemic index than the plain BW and also showed the best scores for sensory attributes. This research confirms the effectiveness of viscous (1→3,1→4)-β-glucan in reducing postprandial blood glucose levels, even in foods with a high glycemic index. The enrichment technique, and water extraction/freeze-drying technique, could enable the use of barley as a source of a high-value fibre for reducing the glycemic index of traditional wheat-based foods such as bread, without negatively affecting their sensory characteristics.     

Wheat Bread Quality as Influenced by the Substitution of Waxy and Regular Barley Flours in Their Native and Extruded Forms

The substitution of wheat flour with barley flour (i.e. native or pretreated/extruded) reduced the loaf volume. Depending on the barley variety and flour pretreatments, the colour and firmness/texture of the bread loaves were altered. Amongst the barley breads prepared from native flours (at 15% barley flour substitution level), Phoenix had higher loaf volume and lower crumb firmness than Candle. However, amongst the barley breads prepared from extruded flours, CDC-Candle had higher loaf volume and lower crumb firmness than Phoenix. The lower loaf volume and firmer crumb texture of barley breads as compared with wheat bread may be attributed to gluten dilution. Also, the physicochemical properties of barley flour components, especially that of β-glucan, can affect bread volume and texture. β-glucan in barley flour, when added to wheat flour during bread making, could tightly bind to appreciable amounts of water in the dough, suppressing the availability of water for the development of the gluten network. An underdeveloped gluten network can lead to reduced loaf volume and increased bread firmness. Furthermore, in yeast leavened bread systems, in addition to CO₂, steam is an important leavening agent. Due to its high affinity for water, β-glucan could suppress the amount of steam generated, resulting in reduced loaf volume and greater firmness. In the present study, breads made with 15% HTHM CDC-Candle flour had highly acceptable properties (loaf volume, firmness and colour) and it indicated that the use of extruded barley flours would be an effective way to increase the dietary fibre content of barley breads.     

Identification of quantitative trait loci for β-glucan concentration in barley grain

The amount of (1 → 3),(1 → 4)-β-d glucan (β-glucan) accumulated in cell walls of barley (Hordeum vulgare L.) kernel is an important determinant for grain end-use. Grain β-glucan concentration is affected by environmental and genetic factors and usually varies from 3 to 6%. In this study, we have analyzed the β-glucan trait in a doubled-haploid (DH) population of 170 lines grown in three separate field trials. Most of the DH lines showed β-glucan values that ranged from that of the low β-glucan parent (cultivar CDC Bold; 3.3%) to that of the high β-glucan parent (breeding line TR251; 5.4%). Eighty-eight lines of the DH population were genotyped using simple sequence repeat (SSR), amplified fragment length polymorphism (AFLP) and diversity array technology (DArT) markers, which were subsequently integrated into a barley genetic map spanning 1059 cM. Interval mapping and multiple-QTL-mapping (MQM) of quantitative trait loci (QTL) from the three trials indicated seven genomic regions associated with low grain β-glucan concentration. For all putative QTLs, the low β-glucan concentration was contributed by alleles from CDC Bold except for two loci on chromosomes 5H that were derived from TR251. A major QTL located to the centromere region of chromosome 7H was identified by both mapping methods for all three trials. The 7H QTL explained up to 39% of the β-glucan concentration and genetic markers associated with the locus may be used to aid selection of high and low β-glucan barley lines.     

Suitability of tef varieties in mixed wheat flour bread matrices: A physico-chemical and nutritional approach

Wheat flour replacement from 0 to 40% by single tef flours from three Ethiopian varieties DZ-01-99 (brown grain tef), DZ-Cr-37 (white grain tef) and DZ-Cr-387 (Quncho, white grain tef) yielded a technologically viable ciabatta type composite bread with acceptable sensory properties and enhanced nutritional value, as compared to 100% refined wheat flour. Incorporation of tef flour from 30% to 40% imparted discreet negative effects in terms of decreased loaf volume and crumb resilience, and increase of crumb hardness in brown tef blended breads. Increment of crumb hardness on aging was in general much lower in tef blended breads compared to wheat bread counterparts, revealing slower firming kinetics, especially for brown tef blended breads. Blended breads with 40% white tef exhibited similar extent and variable rate of retrogradation kinetics along storage, while brown tef-blended breads retrograded slower but in higher extent than control wheat flour breads. Breads that contains 40% tef grain flour were found to contain five folds (DZ-01-99, DZ-Cr-387) to 10 folds (DZ-Cr-37) Fe, three folds Mn, twice Cu, Zn and Mg, and 1.5 times Ca, K, and P contents as compared to the contents found in 100% refined wheat grain flour breads. In addition, suitable dietary trends for lower rapidly digestible starch and starch digestion rate index were met from tef grain flour fortified breads.     

Studies on the baking properties of wheat: Pigeonpea flour blends

Pigeonpea flour was substituted at levels of 0, 5, 10, 15,20, 25% to wheat flour and whole wheat meal for bread andChapatti making, respectively. Blends were prepared up to50% for cookie making. Increasing levels of pigeonpeas inthe blends significantly increased the protein and mineralcontent of the baked products. The bread from 10%pigeonpea flour blend with 2–3% vital gluten and 0.5%SSL had high loaf volume and loaf quality. Blends containing 15% pigeonpea flour were acceptable for Chapatti and 30% pigeonpea flour with 0.25% SSL wereacceptable for cookie making.     

Improving the quality of nutrient-rich Teff (Eragrostis tef) breads by combination of enzymes in straight dough and sourdough breadmaking

The growing interest in the benefits of wholegrain products has resulted in the development of baked products incorporating less utilised and ancient grains such as, millet, quinoa, sorghum and teff. However, addition of wholegrains can have detrimental effects on textural and sensory bread product qualities.Enzymes can be utilised to improve breadmaking performance of wholegrain flours, which do not possess the same visco-elastic properties as refined wheat flour, in order to produce a healthy and consumer acceptable cereal product.The effects of Teff grain on dough and bread quality, selected nutritional properties and the impact of enzymes on physical, textural and sensory properties of straight dough and sourdough Teff breads were investigated.Teff breads were prepared with the replacement of white wheat flour with Teff flour at various levels (0%, 10%, 20%, and 30%) using straight dough and sourdough breadmaking. Different combinations of enzymes, including xylanase and amylase (X + A), amylase and glucose oxidase (A + GO), glucose oxidase and xylanase (GO + X), lipase and amylase (L + A) were used to improve the quality of the highest level Teff breads. A number of physical, textural and sensory properties of the finished products were studied. The nutritional value of breads was determined by measuring chemical composition for iron, total antioxidant capacity, protein, fibre and fat. The obtained results were used to estimates intakes of nutrients and to compare them with DRIs.The incorporation of Teff significantly (P < 0.05) improved dietary iron levels as 30% Teff breads contained more than double the amount of iron when compared to corresponding wheat bread (6 mg/100 g v 2 mg/100 g). Addition of Teff also significantly (P < 0.05) improved total antioxidant capacity from 1.4 mM TEAC/100 g to 2.4 mM TEAC/100 g. It was estimated that an average daily allowance of 200 g of Teff enriched bread would contribute to DRIs in the range of 42-81% for iron in females, 72-138% for iron in males; 38-39% for protein in males, 46-48% for protein in females; and 47-50% of fibre in adults.The major challenge was encountered in producing the highest level of Teff bread with good textural and sensory attributes. Increasing the level of Teff significantly (P < 0.05) increased dough development time, degree of softening, crumb firmness and bitter flavour whilst decreasing the dough stability, specific loaf volume and overall acceptability of the bread. Teff breads produced with the addition of enzyme combinations showed significant improvements (P < 0.05) in terms of loaf volume, crumb firmness, crumb structure, flavour and overall acceptability in both straight dough and sourdough breadmaking.     

Endogenous arabinoxylans variability in refined wheat flour and its relationship with quality traits

Arabinoxylans (AXs) are the major dietary fiber (DF) component in wheat and their consumption has been associated with several health benefits. Genetic improvement of the AX in refined wheat flour could be a good solution to improve the DF daily consumption while maintaining the flour desirable quality. In this study, 193 common wheat lines were analyzed for their AX content in refined flour and end-use quality. Wide variation in both the total arabinoxylan (TOT-AX) (10.8–16.5 mg/g) and water-extractable arabinoxylan (WE-AX) (3.2–7.6 mg/g) was identified and, in both cases, the genotype had the greatest impact on the observed phenotypes. Variation in the endogenous AX fractions appeared to have a moderate effect on wheat quality. The WE-AX, specifically, were positively correlated with gluten strength (r = 0.11 to 0.32) and bread loaf volume (r = 0.16), whereas the TOT-AX were negatively correlated with dough extensibility (r = −0.11) and bread making quality (r = −0.11). Overall, results of this study show that the genetic improvement of grain AX is feasible and that the AXs present in refined flour do not dramatically alter wheat quality indicating that it is possible to select varieties with high AX endosperm content end desired end-use quality.     

Cereal β-glucans in diet and health

The native mixed linkage β-glucan of cereals is classified as a soluble dietary fibre, with rheological properties generally similar to guar gum and other random coil polysaccharides. The ability of oat and barley products to attenuate postprandial glycemic and insulinemic response is related to content of (1→3)(1→4)-β-d-glucan (β-glucan) and viscosity. A role of viscosity of β-glucan has not been directly demonstrated for lowering of serum cholesterol levels, and not all studies report a statistically significant lowering. The wide range of effectiveness reported may partially be explained by the properties of the β-glucan in the diets used, as well as the dose.     

Glucose and pyranose oxidase improve bread dough stability

When used in bread dough systems, glucose oxidase (GO) and pyranose oxidase (P₂O) generate H₂O₂ from O₂. We here studied their potential to improve dough and bread characteristics. Neither GO nor P₂O significantly affected the volume of straight dough bread produced with fermentation and proofing times of respectively 90 and 36 min at dosages up to 0.50 nkat/g flour. Supplementation with 1.00 nkat/g flour of GO or P₂O significantly decreased bread loaf volume. The resistance of dough (fermented for 20 min and proofed for 56 min) to an applied shock was substantially improved by inclusion of 0.08, 0.25, 0.50 or 1.00 nkat/g flour of GO or P₂O in the dough recipe. Thus, the proofed doughs showed significantly less collapse and the resultant breads had higher loaf volumes than did the reference breads. Yeast probably exerts an oxidizing effect on dough, which, depending on the exact breadmaking protocol used, might veil the positive oxidizing effect of the enzymes on dough properties during prolonged fermentation.     

Effect of barley and oat flour types and sourdoughs on dough rheology and bread quality of composite wheat bread

The potential of sourdough to improve bread quality of barley and oat enriched wheat breads may depend on the characteristics of the added flour (cereal type, variety, extraction rate). We compared the effect of different barley flours and oat bran (substitution level 40%), unfermented and as sourdoughs (20% of total flour), on composite wheat dough and bread characteristics by combining empirical rheological analyses (DoughLab, SMS/Kieffer Dough and Gluten Extensibility Rig) with small-scale baking of hearth loaves. Whole grain barley flour sourdough increased resistance to extension (Rmax) of the dough and improved the form ratio of hearth loaves compared to unfermented whole grain barley flour. However, sourdough showed little effect on the breads prepared with sifted barley flour or oat bran. The breads made with oat bran showed highest bread volume, lowest crumb firmness and highest β-glucan calcofluor weight average molecular weight (MW). The heat treatment of oat bran inactivated endogenous enzymes resulting in less β-glucan degradation. High MW β-glucans will increase the viscosity of the doughs water phase, which in turn may stabilise gas cells and may therefore be the reason for the higher bread volume of the oat bran breads observed in our study.     

Sourdough fermentation of wholemeal wheat bread increases solubility of arabinoxylan and protein and decreases postprandial glucose and insulin responses

Glycemic responses to most of the conventional breads are high, including breads made of wholemeal flour. Baking technology is known to affect these responses. The aim of the present study was to investigate effects of xylanase enzyme treatment and sourdough fermentation in wholemeal wheat bread baking on postprandial glucose and insulin responses and on in vitro protein digestibility. The wheat breads were made of 100% flour from peeled kernels by a straight dough or sourdough fermentation method, and with or without using xylanase during mixing of dough. Standard white wheat bread was used as a reference. All test bread portions contained 50 g available carbohydrate and were served in random order to eleven insulin resistant subjects. Blood samples for measuring glucose and insulin concentrations were drawn over 4 h. The sourdough wholemeal wheat bread resulted in the lowest postprandial glucose and insulin responses among the four tested breads (treatment × time; p = 0.000 and p = 0.022, respectively). There were differences in solubility and depolymerisation of protein and arabinoxylan among the breads but these did not fully explain the in vivo findings. In conclusion, the health effects of wholemeal wheat bread can be further improved by using sourdough process in breadmaking.     

Effects of sulphur on yield and malting quality of barley

Eight field experiments were conducted at four sites in the UK in 2003 and 2004 to investigate the effects of sulphur (S) application on yield and malting quality of barley. Significant yield responses to S additions were obtained in five out of the eight experiments, with yield increases ranging from 0.2 to 1.2 t/ha (4.7–22.5%). At the two most S-deficient sites, S application significantly increased malt diastatic power, alpha-amylase activity, friability and homogeneity, and decreased (1→3,1→4)-β-glucan concentration in the wort, indicating an improved endosperm modification during malting. Sulphur applications also significantly increased the concentration of S-methylmethionine (the precursor of dimethylsulphide) in kilned malt, which could impact on beer flavour. When the supply of N was limiting, S applications decreased grain N concentration due to a dilution effect as a result of increased grain yield. In some cases, S applications resulted in decreased grain size. At sites non-deficient or marginally deficient in S, applications of S had little effect on grain or malting quality parameters. The need to maintain an adequate S supply to barley for both yield and malting quality was demonstrated.     

A Novel α4/7-Conotoxin QuIA Selectively Inhibits α3β2 and α6/α3β4 Nicotinic Acetylcholine Receptor Subtypes with High Efficacy

α6β4 nAChR is expressed in the peripheral and central nervous systems and is associated with pain, addiction, and movement disorders. Natural α-conotoxins (α-CTxs) can effectively block different nAChR subtypes with higher efficacy and selectivity. However, the research on α6β4 nAChR is relatively poor, partly because of the lack of available target-specific α-CTxs. In this study, we synthesized a novel α-4/7 conotoxin QuIA that was found from Conus quercinus. We investigated the efficacy of this peptide to different nAChR subtypes using a two-electrode voltage-clamp technique. Remarkably, we found α-QuIA inhibited the neuronal α3β2 and α6/α3β4 nAChR subtypes with significantly high affinity (IC₅₀ was 55.7 nM and 90.68 nM, respectively), and did not block other nAChR subtypes even at a high concentration of 10 μM. In contrast, most α-CTxs have been determined so far to effectively block the α6/α3β4 nAChR subtype while also maintaining a similar higher efficacy against the closely related α6β2β3 and/or α3β4 subtypes, which are different from QuIA. In conclusion, α-QuIA is a novel α4/7-CTx, which has the potential to develop as an effective neuropharmacology tool to detect the function of α6β4 nAChR.     

Effect of barley flour addition on the physico-chemical properties of dough and structure of bread

The effects of different percentages of barley flour (i.e. 0–25%) in wheat flours on the physico-chemical properties and structure of dough and bread were investigated. As the percentage of barley flour in mixed flour was increased, its protein and gluten contents decreased whereas the ash content and enzyme activity increased. The rheological characteristics of the four dough mixes were studied using Farinograph, Extensograph and Alveograph. The water absorption (p < 0.01) and stability (p < 0.05) decreased significantly as the percentage of barley flour increased, while no changes were observed in the extensibility and maximum heights. Significant differences were observed in the structural and physical properties as well as in the image analysis of breads. With the increase in the percentage of barley flour, the crumb apparent density decreased (p < 0.1) whereas the porosity (i.e., fraction to total volume) increased (p < 0.1). Overall, the shape and pore structure at 10% barley flour (W90B10) were similar to the pure wheat flour bread, while addition at 15 and 25% of barley flour (W85B15 and W75B25) showed more non-uniform and larger pores.     

Molecular weight distribution and content of water-extractable β-glucan in rye crisp bread

Rye crisp doughs and breads were made from three rye flours with different falling numbers (i.e. different enzyme activity), with or without oat bran rich in β-glucan (OBG) added, and with different fermentation times. The aim of the study was to investigate the effect of endogenous enzymes in the flour on molecular weight and content of water-extractable β-glucan in the doughs and breads. The molecular weight distribution of β-glucan was unimodal for the flours, with a calcofluor average molecular weight of about 90×10⁴ g/mol for the rye flours, and 150×10⁴ g/mol for rye flours with added OBG. The molecular weight decreased with increasing fermentation time for all doughs, but was almost unchanged during oven-baking. The calcofluor average molecular weight was highest in doughs and breads made of flour with highest falling number (i.e. lowest enzyme activity), indicating that the endogenous enzymes in the flour were responsible for the breakdown of β-glucan. During fermentation, water-extractable β-glucan was released from the insoluble matrix, probably by the endogenous enzymes in the flour. When the molecular weight of the water-extractable β-glucan decreased, they probably associated with each other or with other components in the dough to form unextractable complexes. The content of water-extractable β-glucan did not change during oven-baking.     

Cultivar and Environmental Effects on (1→3,1→4)-β-D-Glucan and Protein Content in Malting Barley

A low β-glucan and protein content is desirable for malting barley. In China high β-glucan content has been considered a major factor contributing to the inferior malting quality of local barleys. In this study 10 barley cultivars were planted in multi-location trials in the southern winter-barley zone to determine genotypic and environmental effects on β-glucan and protein content. There were highly significant differences in both β-glucan and protein content between the 10 barleys, the eight locations and the 2 planting years. The average β-glucan content over the 2 years for the 10 cultivars in the eight locations ranged from 3·91% for Gangpi 1 to 4·95% for Xiumei 3, and from 3·76% for Hangzhou to 4·75% for Taian in eight locations. Correspondingly, the protein contents of the 10 cultivars ranged from 11·37% for Yanyin 1 to 12·52% for ZAU, and of the eight locations from 9·5% for Hangzhou to 14·69% for Taian. Environmental factors contributed the largest component of the variation in both β-glucan and protein content. Five climatic factors, namely the total accumulated temperature, accumulated temperature to 25 °C and to 30 °C, precipitation and days with rain during seed development were included in a regression equation relating climatic factors and β-glucan content. In the regression equation relating climatic factors and protein content, two other factors, days from heading to maturity and accumulated temperature to 20 °C were included, in addition to the five climatic factors that significantly affected β-glucan content.     

Interactions of Globular and Ribbon [γ4E]GID with α4β2 Neuronal Nicotinic Acetylcholine Receptor

The α4β2 nAChR is implicated in a range of diseases and disorders including nicotine addiction, epilepsy and Parkinson’s and Alzheimer’s diseases. Designing α4β2 nAChR selective inhibitors could help define the role of the α4β2 nAChR in such disease states. In this study, we aimed to modify globular and ribbon α-conotoxin GID to selectively target the α4β2 nAChR through competitive inhibition of the α4(+)β2(−) or α4(+)α4(−) interfaces. The binding modes of the globular α-conotoxin [γ4E]GID with rat α3β2, α4β2 and α7 nAChRs were deduced using computational methods and were validated using published experimental data. The binding mode of globular [γ4E]GID at α4β2 nAChR can explain the experimental mutagenesis data, suggesting that it could be used to design GID variants. The predicted mutational energy results showed that globular [γ4E]GID is optimal for binding to α4β2 nAChR and its activity could not likely be further improved through amino-acid substitutions. The binding mode of ribbon GID with the (α4)₃(β2)₂ nAChR was deduced using the information from the cryo-electron structure of (α4)₃(β2)₂ nAChR and the binding mode of ribbon AuIB. The program FoldX predicted the mutational energies of ribbon [γ4E]GID at the α4(+)α4(−) interface, and several ribbon[γ4E]GID mutants were suggested to have desirable properties to inhibit (α4)₃(β2)₂ nAChR.