Effect of Extrusion Cooking on the Primary Structure and Water Solubility of β‐Glucans from Regular and Waxy Barley

Water‐soluble β‐glucan from native and extrusion‐cooked barley flours of two barley cultivars, Candle (a waxy starch barley) and Phoenix (a regular starch barley), was isolated and purified. The purity of β‐glucan samples was 85–93% (w/w, dry weight basis) for Candle and 77–86% (w/w, dry weight basis) for Phoenix. The water solubility of β‐glucan (at room temperature, 25°C) in the native and extruded flours (primary solubility) was different from that of the purified β‐glucan samples (secondary solubility). The solubility of β‐glucan in the native and extruded Candle flour was substantially higher than that of β‐glucan in Phoenix. For both cultivars, β‐glucan in the extruded flours had solubility (primary solubility) values higher than in their native counterparts. The solubility of β‐glucan in the purified β‐glucan samples differed depending on the barley cultivar and the extrusion conditions employed. The glycosidic linkage profiles of purified soluble β‐glucan from native and extruded barley flours were determined in order to understand the changes in the primary structure of β‐glucan and the effect of extrusion on the β‐glucan structure‐solubility relationship.     

Extraction of β‐Glucan from Oats for Soluble Dietary Fiber Quality Analysis

Extraction protocols for β‐glucan from oat flour were tested to determine optimal conditions for β‐glucan quality testing, which included extractability and molecular weight. We found mass yields of β‐glucan were constant at all temperatures, pH values, and flour‐to‐water ratios, as long as sufficient time and enough repeat extractions were performed and no hydrolytic enzymes were present. Extracts contained about 30–60% β‐glucan, with lower proportions associated with higher extraction temperatures in which more starch and protein were extracted. All commercial starch hydrolytic enzymes tested, even those that are considered homogenous, degraded β‐glucan apparent molecular weight as evaluated by size‐exclusion chromatography. Higher concentration β‐glucan solutions could be prepared by controlling the flour‐to‐water ratio in extractions. Eight grams of flour per 50 mL of water generated the highest native β‐glucan concentrations. Routine extractions contained 2 g of enzyme‐inactivated flour in 50 mL of water with 5mM sodium azide (as an antimicrobial), which were stirred overnight, centrifuged, and the supernatant boiled for 10 min. The polymer extracted had a molecular weight of about 2 million and was stable at room temperature for at least a month.     

Determination of β‐Glucan Molecular Weight Using SEC with Calcofluor Detection in Cereal Extracts

A high‐performance size‐exclusion chromatography system (HPSEC) was set up with detection based on the specific binding of Calcofluor to β‐glucan for determination of amount and molecular weight of β‐glucan in different cereal extracts. To calibrate the HPSEC system, a purified β‐glucan was fractionated into narrow molecular weight ranges and the average molecular weight was determined before analysis on the HPSEC system. The detector response was similar for β‐glucans from oats and barley and appeared to be independent of molecular weight. Four different methods for extraction of β‐glucan from different cereal products were tested: two alkaline, one with hot water and added α‐amylase, and one with water and added xylanase. Inactivation of endogenous β‐glucanase was crucial for the stability of the extracts, even when extracting at high temperature or pH. Yields varied widely between the different extraction methods but average molecular weight and molecular weight distribution were similar. Extraction with sodium hydroxide generally gave a higher yield and molecular weight of β‐glucan in the extracts.     

Roller Milling of Canadian Hull‐less Barley: Optimization of Roller Milling Conditions and Composition of Mill Streams

Roller milling of hull‐less barley generates mill streams with highly variable β‐glucan and arabinoxylan (AX) content. For high β‐glucan cultivars, yields >20% (whole barley basis) of a fiber‐rich fraction (FRF) with β‐glucan contents >15% can be readily obtained with a simple short mill flow. Hull‐less barley cultivars with high β‐glucan content require higher power consumption during roller milling than normal β‐glucan barley. Recovery of flour from high β‐glucan cultivars was greatly expedited by impact passages after grinding, particularly after reduction roll passages. Pearling before roller milling reduces flour yield and FRF yield on a whole unpearled barley basis, but flour brightness is improved and concentration of β‐glucan in fiber‐rich fractions increases. Pearling by‐products are rich in AX. Pearling to 15–20% is the best compromise between flour and FRF yield and flour brightness and pearling by‐products AX content. Increasing conditioning moisture from 12.5 to 14.5% strongly improved flour brightness with only a moderate loss of flour yield on a whole unpearled barley basis. As moisture content was increased to 16.5%, flour yield declined without a compensating improvement in brightness, but the yield of fiber‐rich fraction continued to increase and concentration of β‐glucan in FRF also increased.     

Network Formation by Pilot Plant and Laboratory‐Extracted Barley β‐Glucan and Its Rheological Properties in Aqueous Solutions

Barley and oat β‐glucans of low viscosity form reversible gels when prepared in sufficiently high concentrations. Solutions of three barley β‐glucan gums differing in molecular weight and thus in viscosity were prepared at 1.0, 2.5, or 5.0% (w/w) concentration levels. Medium‐ and high‐viscosity gums were prepared in a pilot plant (PP) and laboratory (LAB), respectively. Low‐viscosity (LV) gum was extracted in the laboratory at pH 7, which allowed for native enzymatic activity and decreased molecular weight. Network formation was monitored overnight through changes in storage (G′) and loss (G″) moduli. The strength of the formed network was determined from oscillatory rheological measurements by increasing the strain from 2 to 100%. Findings demonstrate that gelation of β‐glucan is molecular weight dependent and practically an instantaneous process for low‐viscosity gum solutions at concentrations of ≤5% gum (or ≤4% β‐glucan), levels lower than previously anticipated. The purity of β‐glucan also seems to affect gelation rate. Better understanding of the β‐glucan gelation behavior is important for its functionality in both food product applications and physiological mechanisms of its health benefits.     

Distribution of β‐Glucan in the Grain of Hull‐less Barley

Nine hull‐less barley (HB) containing waxy (0–7% amylose), normal (≈25% amylose), or high amylose (≈42% amylose) starch with normal or fractured granule make‐up and 4–9% (1→3)(1→4)‐β‐d ‐glucans (β‐glucan) were pearled to remove 70% of the original grain weight in 10% intervals. The pearled fractions were analyzed for β‐glucan distribution within HB grain. Protein content of the pearled fractions indicated that the three outermost fractions contained pericarp and testa, aleurone, and subaleurone tissues, respectively. For all HB, β‐glucan and acid‐extract viscosity were very low in the outermost 20% of the kernel. For low β‐glucan HB, β‐glucan content was the greatest in the subaleurone region and declined slightly toward inner layers. For high β‐glucan HB, however, more than 80% of grain β‐glucan was distributed more evenly throughout the endosperm. Acid extract viscosity was significantly (P < 0.01) correlated with total (r = 0.75) and soluble (r = 0.87) β‐glucan content throughout the kernel of all HB. Growing conditions, location and year, had significant effects on the concentration of protein, starch and β‐glucan. However, protein, starch, and β‐glucan distribution patterns were not affected by growing conditions. The difference in β‐glucan distribution between low and high β‐glucan HB may explain the difference in milling performance of HB with low or high β‐glucan.     

Effects of β‐Glucan Content and Pearling of Barley in Diet‐Induced Obese Mice

The effects of the β‐glucan content and pearling of barley on abdominal obesity and the proinflammatory state were investigated in diet‐induced obese mice. Male C57BL/6J mice were randomly divided into four groups and fed either a high‐fat diet containing high‐β‐glucan barley (Beau Fiber [BF]) or a high‐fat diet containing β‐glucan‐free barley (Shikoku‐hadaka 84(bgl ) [BGL]) as whole grain flour or 60% pearled flour for 12 weeks. The weights of mesenteric fat, serum total and low density lipoprotein cholesterol levels, serum insulin and fasting glucose levels, oral glucose tolerance test results, and messenger RNA (mRNA) expression of proinflammatory markers in epididymal fat in both BF groups were significantly lower than those of both BGL groups. The abundance of Bacteroides in both BF groups was significantly higher than that in both BGL groups, whereas the abundance of Clostridium clusters in both BF groups was significantly lower than that in both BGL groups. No significant differences between the whole grain and pearled flours were observed. These results suggest that high‐β‐glucan barley attenuates the progression of abdominal obesity and the proinflammatory state in diet‐induced obese mice compared with β‐glucan‐free barley, possibly by modifying insulin secretion and the microbiota.     

Composition and Utilization of Barley Pearling By‐Products for Making Functional Pastas Rich in Dietary Fiber and β‐Glucans

Pearling by‐products and the pearled products of two commercial stocks of hulled barley, pearled according to an industrial process consisting of five consecutive pearling steps, were analyzed for β‐glucans, dietary fiber (total, soluble, and insoluble), protein, lipid, ash, and digestible carbohydrate. The data showed that the pearling flour fractions, abraded in the fourth and fifth hullers, contained interesting amounts of β‐glucans (3.9–5.1% db) from a nutritional point of view. These fractions were subsequently enriched in β‐glucans using a milling‐sieving process to double β‐glucan content (9.1–10.5% db). Functional pastas, enriched with β‐glucans and dietary fiber, were produced by substituting 50% of standard durum wheat semolina with β‐glucan‐enriched barley flour fractions. Although darker than durum wheat pasta, these pastas had good cooking qualities with regard to stickiness, bulkiness, firmness, and total organic matter released in rinsing water. The dietary fiber (13.1–16.1% wb) and β‐glucan (4.3–5.0% wb) contents in the barley pastas were much higher than in the control (4.0 and 0.3% wb, respectively). These values amply meet the FDA requirements of 5 g of dietary fiber and 0.75 g of β‐glucans per serving (56 g in the United States and 80 g in Italy). At present, the FDA has authorized the health claim “may reduce the risk of heart disease” for food containing β‐glucans from oat and psyllium only.     

Extraction and Physicochemical Characterization of Rye β‐Glucan and Effects of Barium on Polysaccharide Molecular Weight

Use of saturated Ba(OH)₂ to extract rye β‐glucan led to a depolymerized product. Similar depolymerization of β‐glucan was observed when oat bran was extracted with this reagent. Isolated oat β‐glucan, detarium xyloglucan, guar galactomannan, and wheat and rye arabinoxylan were also depolymerized by treatment with the barium reagent. The degree of depolymerization was related to time of contact with, and concentration of, the barium. Rye β‐glucan of two different molecular weights (MW) were isolated and characterized. The structure of rye β‐glucan, as evaluated from the ratio of (1→3)‐linked cellotriosyl to (1→3)‐linked cellotetraosyl primary structural units, most closely resembles barley β‐glucan. Analytical variability of this ratio is discussed. A freshly prepared solution (2%) of the higher MW sample showed shear thinning behavior typical of cereal β‐glucans. The lower MW sample at 2% was not shear thinning, but on further purification, after storage for seven days, a 6% solution had gelled as shown by the mechanical spectrum.     

Isolation,Fractionation, and Structural Characteristics of Alkali‐Extractable β‐Glucan from Rye Whole Meal

The main nonstarch polysaccharide of rye is arabinoxylan (AX), but rye contains significant levels of (1→3)(1→4)‐β‐d ‐glucan, which unlike oat and barley β‐glucan, is not readily extracted by water, possibly because of entrapment within a matrix of AX cross‐linked by phenolics. This study continues objectives to improve understanding of factors controlling the physicochemical behavior of the cereal β‐glucans. Rye β‐glucan was extracted by 1.0N NaOH and increasing concentrations of ammonium sulfate were used to separate the β‐glucan from AX and prepare a series of eight narrow molecular weight (MW) distribution fractions. Composition and structural characteristics of the isolated β‐glucan and the eight fractions were determined. High‐performance size‐exclusion chromatography (HPSEC) with both specific calcofluor binding and a triple detection (light scattering, viscometry, and refractive index) system was used for MW determination. Lichenase digestion followed by high‐performance anion exchange chromatography of released oligosaccharides, was used for structural evaluation. The overall structure of all fractions was similar to that of barley β‐glucan.     

Milling Performance of North European Hull‐less Barleys and Characterization of Resultant Millstreams

Four hull‐less barley samples were milled on a Bühler MLU 202 laboratory mill and individual and combined milling fractions were characterized. The best milling performance was obtained when the samples were conditioned to 14.3% moisture. Yields were 37–48% for straight‐run flour, 47–56% for shorts, and 5–8% for bran. The β‐glucan contents of the straight‐run white flours were 1.6–2.1%, of which ≈49% was water‐extractable. The arabinoxylan contents were 1.2–1.5%, of which ≈17% was water‐extractable. Shorts and bran fractions contained more β‐glucan (4.2–5.8% and 3.0–4.7%, respectively) and arabinoxylan (6.1–7.7% and 8.1–11.8%, respectively) than the white flours. For those fractions, β‐glucan extractability was high (58.5 and 52.3%, respectively), whereas arabinoxylan extractability was very low (≈6.5 and 2.0%, respectively). The straight‐run white flours had low α‐amylase, β‐glucanase, and endoxylanase activities. The highest α‐amylase activity was found in the shorts fractions and the highest β‐glucanase and endoxylanase activities were generally found in the bran fractions. Endoxylanase inhibitor activities were low in the white flours and highest in the shorts fractions. High flavanoid, tocopherol, and tocotrienol contents were found in bran and shorts fractions.     

Molecular Weight of β‐Glucan Affects Physical Characteristics,In Vitro Bile Acid Binding,and Fermentation of Muffins

Muffins containing different amounts and molecular weights (MW) of β‐glucan were evaluated for the effect of β‐glucan on the physical characteristics of the muffins and on in vitro bile acid binding and fermentation with human fecal flora. Wheat flour muffins were prepared with the addition of β‐glucan extracts with high‐, medium‐, or low‐MW. For oat flour muffins, the native oat flour contained high‐MW β‐glucan; the oat flours were treated to create medium‐ and low‐MW β‐glucan within the prepared muffin treatments. For each 60‐g muffin, the amounts of β‐glucan were 0.52, 0.57, and 0.59 g for high‐, medium‐, and low‐MW β‐glucan wheat flour muffins, and 2.38, 2.18, and 2.23 g for high‐, medium‐, and low‐MW β‐glucan oat flour muffins, respectively. The lower the MW of the β‐glucan in muffins, the lower the height and volume of the muffins. The oat flour muffins were less firm and springy than the wheat flour muffins as measured on a texture analyzer; however, MW had no effect on muffin texture. The oat flour muffins bound more bile acid than did the wheat flour muffins. The muffins with high‐MW β‐glucan bound more bile acid than did those with low‐ and medium‐MW β‐glucan. Muffin treatment affected the formation of gas and total short‐chain fatty acids (SCFA) compared with the blank without substrate during in vitro fermentation. There were no differences in pH changes and total gas production among muffin treatments. The high‐MW β‐glucan wheat flour muffins produced greater amounts of SCFA than did the wheat flour muffin without β‐glucan and the oat flour muffins; however, there were no differences in SCFA production among muffins with different MW. In general, the β‐glucan MW affected the physical qualities of muffins and some potential biological functions in humans.     

REVIEW: Oat and Rye β‐Glucan: Properties and Function

Over the years, the β‐glucan of oats and barley has been the subject of study either because of the importance of the cholesterol‐lowering potential to health claims (FDA 1997, 2005) or, in the case of barley, because of the role of β‐glucan and β‐glucan‐rich endosperm cell walls in malting and brewing. β‐Glucan is also present in rye and in much lesser amounts in wheat. The most striking difference in these latter two sources is the difficulty in extractability; alkali rather than water is required for significant release from the cell walls. This review will discuss physicochemical properties of oat and rye β‐glucan and, where information allows, relate these to physiological effects. Viscosity, or more generally rheology, plays a central role in discussions of cereal β‐glucan functionality and physiological effects and will be the focus of this review.     

Release of β‐Glucan from Cell Walls of Starchy Endosperm of Barley

β‐Glucan can be solubilized from barley by warm water, with increasing solubilization as the temperature is increased. Substantially less glucan is extracted if the barley is dehusked using sulfuric acid, particularly if the dehusked barley is denatured. This indicates that enzymes capable of solubilizing glucan are present in barley. Various purified enzymes promote the solubilization of glucan from denatured and dehusked barley. Apart from endo‐β‐(1→3)(1→4)‐glucanase, these enzymes include endo‐xylanases, arabinofuranosidase, xyloacetylesterase, and feruloyl esterase. Ferulic acid and, probably, acetyl groups are esterlinked to arabinoxylan, not β‐glucan, in the cell walls of barley starchy endosperm, so the ability of the esterases, xylanases, and arabinofuranosidase to solubilize glucan indicates the pentosan component of the cell wall can restrict the extraction of glucan.     

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.     

Comparison of Different Types of NIR Instruments in Ability to Measure β‐Glucan Content in Naked Barley

Importance of β‐glucan in human nutrition is mirrored in numerous approval applications registering β‐glucan containing products as health beneficial products in accordance with forthcoming EU Health Claims Regulation. In comparison to other cereals, barley contains considerable amounts of β‐glucan. Naked barley is of particular interest because it circumvents the costs and loss of beneficial substances related to dehusking. In this study, the potential of near‐infrared spectroscopy as an accurate, fast and economic method of determination of β‐glucan in naked barley was appraised. Four different near‐infrared instruments were used to analyze 107 barley samples, in both whole grain and milled form. Importantly, both black and purple pericarp samples, which are of additional nutritional interest due to high anthocyanin content, and waxy samples, which show an extraordinary high β‐glucan content could be analyzed within the same calibration set as the normal samples. All tested dispersive near‐infrared reflection instruments showed suitability for supervision of breeding experiments and β‐glucan monitoring in food industries (R² > 0.78). Common, industrially used near‐infrared transmission instruments also provided reasonable results, although only suitable for rough selection according to β‐glucan levels. On the other hand, the Fourier transform near‐infrared reflection instrument was able to perform analytical analyses (R² = 0.96–0.98).     

Bulk Carbohydrate Grain Filling of Barley β‐Glucan Mutants Studied by 1H HR MAS NMR

Temporal and genotypic differences in bulk carbohydrate accumulation in three barley genotypes differing in the content of mixed linkage β‐(1→3),(1→4)‐D‐glucan (β‐glucan) and starch were investigated using proton high‐resolution, magic angle spinning, nuclear magnetic resonance (¹H HR MAS NMR) during grain filling. For the first time, ¹H HR MAS NMR spectra of flour from immature barley seeds are analyzed. Spectral assignments are made using two‐dimensional (2D) NMR methods. Both α‐ and β‐glucan biosynthesis were characterized by inspection of the spectra as well as by calibration to the reference methods for starch and β‐glucan content. Starch was quantified with very good calibrations to the α‐(1→4) peak (5.29–5.40 ppm) and the region 3.67–3.83 ppm covering starch glycopyranosidic protons from H5 and H6. In contrast, the spectral inspection of the β‐anomeric region 4.45–4.85 ppm showed unexpected lack of intensity in the high β‐glucan mutant lys5f at seed maturity, resulting in poor calibration to reference β‐glucan content. We hypothesize that the lack of β‐glucan signal in lys5f indicates partial immobilization of the β‐glucan that appears to be either genotypic dependent or water/β‐glucan ratio dependent.     

In Vitro Binding of Bile Acids by Rice Bran,Oat Bran,Barley and β‐Glucan Enriched Barley

The in vitro bile acid binding by rice bran, oat bran, dehulled barley, and β‐glucan enriched barley was determined using a mixture of bile acids at a duodenal physiological pH of 6.3. Six treatments and two blank incubations were conducted testing substrates on an equal protein basis. The relative in vitro bile acid binding of the cereal brans on an equal total dietary fiber (TDF) and insoluble dietary fiber (IDF) basis considering cholestyramine as 100% bound was rice bran 45 and 49%; oat bran 23 and 30%; dehulled barley 33 and 57%; and β‐glucan enriched barley 20 and 40%, respectively. Bile acid bindings on equal protein basis for the respective cereals were 68, 26, 41, and 49%. Bile acid binding by rice bran may account to a great extent for its cholesterol‐lowering properties, while bile acid binding by oat bran suggests that the primary mechanism of cholesterol lowering by oat bran is not due to the bile acid binding by its soluble fiber. Bile acid binding was not proportional to the soluble fiber content of the cereal brans tested. Except for dehulled barley, bile acid binding for rice bran, oat bran, and β‐glucan enriched barley appear to be related to their IDF content. Highest relative bile acid binding values for rice bran and β‐glucan enriched barley were observed on an equal protein basis, whereas highest values for dehulled barley were based on IDF. Data suggest that of all four cereals tested, bile acid binding may be related to IDF or protein anionic, cationic, physical and chemical structure, composition, metabolites, or their interaction with active binding sites.     

Prediction of β‐Glucan Concentration Based on Viscosity Evaluations of Raw Oat Flours from High β‐Glucan and Traditional Oat Lines

Rheological properties of raw oat flour slurries were determined in experimental high β‐glucan (≤7.8%) and traditional oat lines (4–5% β‐glucan) grown in two consecutive years. Three different media were used to disperse oat flours: deionized water, silver nitrate solution (to inactivate endogenous enzymes), and alkali solution (to solubilize both water‐soluble and water‐insoluble β‐glucans). Significant correlations (P < 0.05) between viscosity of slurries and β‐glucan concentration obtained in either deionized water (r = 0.833), silver nitrate (r = 0.940), or alkali (r = 0.896) solutions showed that β‐glucans were the main contributor to oat extract viscosity. The highest correlation was obtained in silver nitrate solution, suggesting that inactivating endogenous enzymes is important to obtain high correlations. Predictive models of oat β‐glucan concentration based on the viscosity profile were developed using partial least squares (PLS) regression. Prediction of β‐glucan concentration based on viscosity was most effective in the silver nitrate solution (r = 0.949, correlation coefficient of predicted vs. analyzed β‐glucans) and least effective in the alkali solution (r = 0.870). These findings demonstrate that the β‐glucan in oat could be predicted by measuring the viscosity of raw flours in silver nitrate solution, and this method could be used as a screening tool for selective breeding.     

Development of an Orange‐Flavored Barley β‐Glucan Beverage

Barley β‐glucan concentrate shows great potential as a functional food ingredient, but few product applications exist. The objectives of this study were to formulate a functional beverage utilizing barley β‐glucan concentrate, and to make a sensory evaluation of beverage quality in comparison to pectin beverages and to assess shelf stability over 12 weeks. Three beverage treatments containing 0.3, 0.5, and 0.7% (w/w) barley β‐glucan were developed in triplicate. Trained panelists found peely‐ and fruity‐orange aroma and sweetness intensity to be similar (P > 0.05) for all beverages tested. Beverage sourness intensity differed among beverages (P ≤ 0.05). Panelists evaluated beverages containing 0.3% hydrocolloid as similar (P > 0.05), whereas beverages with 0.5 and 0.7% β‐glucan were more viscous (P ≤ 0.05) than those with pectin at these levels. Acceptability of beverages was similar according to the consumer panel. Shelf stability studies showed no microbial growth and stable pH for all beverages over 12 weeks. Colorimeter values for most beverages decreased (P ≤ 0.05) during the first week of storage, mostly stabilizing thereafter. With an increase in concentration, β‐glucan beverages became lighter in color (P ≤ 0.05) and cloudier, but these attributes for pectin beverages were not affected (P > 0.05). β‐Glucan beverages exhibited cloud loss during the first three weeks of storage. β‐Glucan can therefore be successfully utilized in the production of a functional beverage acceptable to consumers.