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.     

Molecular Weight Distribution of β‐Glucan in Oat‐Based Foods

Oats, different oat fractions as well as experimental and commercial oat‐based foods, were extracted with hot water containing thermostable α‐amylase. Average molecular weight and molecular weight distributions of β‐glucan in extracts were analyzed with a calibrated high‐performance size‐exclusion chromatography system with Calcofluor detection, specific for the β‐glucan. Oats, rolled oats, oat bran, and oat bran concentrates all had high Calcofluor average molecular weights (206 × 10⁴ to 230 × 10⁴ g/mol) and essentially monomodal distributions. Of the oat‐containing experimental foods, extruded flakes, macaroni, and muffins all had high average molecular weights. Pasteurized apple juice, fresh pasta, and teacake, on the other hand, contained degraded β‐glucan. Calcofluor average molecular weights varied from 24 × 10⁴ to 167 × 10⁴ g/mol in different types of oat bran‐based breads baked with almost the same ingredients. Large particle size of the bran and short fermentation time limited the β‐glucan degradation during baking. The polymodal distributions of β‐glucan in these breads indicated that this degradation was enzymatic in nature. Commercial oat foods also showed large variation in Calcofluor average molecular weight (from 19 × 10⁴ g/mol for pancake batter to 201 × 10⁴ g/mol for porridge). Boiling porridge or frying pancakes did not result in any β‐glucan degradation. These large differences in molecular weight distribution for β‐glucan in different oat products are very likely to be of nutritional importance.     

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.     

Extractability,Structure and Molecular Weight of β‐Glucan from Canadian Rye (Secale cereale L.) Whole Meal

Oat and barley (1→3)(1→4)‐β‐d ‐glucans (β‐glucan) are readily extracted by hot water but rye β‐glucan is resistant to such extraction. This poor extractability might be due to entrapment within a matrix of arabinoxylan (AX) cross‐linked through phenolic constituents. AX are the major nonstarch polysaccharides of the rye kernel. In this study, several approaches were compared in an effort to determine optimum conditions for extraction of high yields, high molecular weight (MW), and high purity of β‐glucan from Canadian rye whole meal. Variables investigated included sodium hydroxide concentrations, extraction time, sample prehydration, extraction under low temperature, and prior extraction of AX with barium hydroxide. There was a linear relationship between the strength of NaOH and amount of β‐glucan extracted and because MW was essentially the same up to 1.0N NaOH, this extraction agent, at room temperature for 90 min, was selected to isolate rye β‐glucan. The β‐glucan was then purified and structure and molecular weight distribution studied.     

Influence of Oat β‐Glucan Preparations on the Perception of Mouthfeel and on Rheological Properties in Beverage Prototypes

The aim was to study the effect of concentration and molecular weight of four different β‐glucan preparations on the perceived sensory quality of a beverage prototype. The correlations between sensory and instrumental measures were investigated. Two of the preparations were brantype containing high molecular weight β‐glucan, two were more‐processed low molecular weight β‐glucan preparations. Twelve beverage samples containing 0.25–2% β‐glucan and one reference sample thickened with carboxymethyl cellulose (CMC) were profiled by a sensory panel and analyzed by instrumental measurements (viscosity and molecular weight). Sensory profiles of the beverages varied at the same concentration of β‐glucan, depending on β‐glucan preparation. Beverages made with the bran‐type preparations were more viscous and had higher perceived thickness than beverages made with more‐processed, low molecular weight preparations. Moderate correlations were obtained between perceived thickness and sliminess and instrumental viscosity at all shear rates between 26 and 100/sec (r = 0.63–0.78; P ≤ 0.001). Technologically, more‐processed β‐glucan preparations are easier to add into a beverage in amounts sufficient for achieving a physiologically functional amount of β‐glucan in a product.     

Detection,Localization, and Variability of Endogenous β‐Glucanase in Wheat Kernels

Clinical studies with isolates of β‐glucan have shown that the health benefits are regulated not only by the polysaccharide concentration but also by the molecular weight and concentration in solution, because these health benefits are controlled, inter alia, by viscosity in the gut. The degradation of β‐glucan in baked products is likely caused by baking ingredients or processes, or by endogenous enzymes in wheat flour. The objectives of the present study were to quantify β‐glucanase in wheat kernels and to determine factors that influence the levels of this enzyme. A modified protocol to quantify β‐glucanase was developed and then confirmed through high‐performance size‐exclusion chromatography (HPSEC) with Calcofluor detection. Under this protocol, it was shown that the concentration of β‐glucanase activity was the highest in the bran fraction of the kernel in ungerminated wheats, whereas it was distributed throughout the entire kernel following germination. Furthermore, investigation on different wheat cultivars planted in the same and different locations showed that genotype, environment, and agronomic practice all can have an effect on β‐glucanase activity level in wheat kernels.     

Preparation and Characterization of Films Using Barley and Oat β‐Glucan Extracts

Films for potential food use were prepared from aqueous solutions of β‐glucan extracted from hulled barley, hull‐less barley, and oats. The extracts (75.2–79.3% β‐glucan) also contained proteins, fat, and ash. Glycerol was used as a plasticizer. The films were translucent, smooth, and homogeneous in structure on both sides. Water vapor permeability of films prepared from 4% solutions of β‐glucan extracts were higher than those from 2% solutions, despite similar values for water vapor transmission rate. Mechanical properties were influenced by both β‐glucan source and concentration. The oat β‐glucan films showed higher tensile strength and water solubility, and lower color, opacity, and deformation values than those of barley. Films prepared from hull‐less barley cv. HLB233 remained intact upon immersion in water for 24 hr.     

Factors Influencing β‐Glucan Levels and Molecular Weight in Cereal‐Based Products

The beneficial role of soluble dietary fiber in human nutrition is well documented and has lead to a growing demand for the incorporation of β‐glucan, particularly from oats and barley, into foods. β‐Glucan with high solubility and high molecular weight distribution results in increased viscosity in the human intestine, which is desirable for increased physiological activity. Molecular weight, level, and solubility of β‐glucan are affected by genotype, environment, agronomic input, and the interactions of these factors and food processing methods. Available literature reveals that the level of β‐glucan in a finished product (e.g. bread, cake, muffins) depends upon several factors in the production chain, whereas food processing operations are major factors affecting molecular weight and solubility of β‐glucans. Therefore, to avail themselves of the natural bioactive compounds, food manufacturers must pay attention not only to ensure sufficient concentration of β‐glucan in the raw material but also to the processing methods and functional properties of β‐glucan, minimizing enzymatic or mechanical breakdown of the β‐glucans in end‐product and optimizing processing conditions. This review discusses the different sources of β‐glucan for use in human functional foods and factors affecting the levels and the molecular weight of β‐glucan at various pre‐ and postharvest operations.     

Content and Molecular Weight Distribution of Oat β‐Glucan in Oatrim,Nutrim, and C‐Trim Products

The soluble fiber, β‐glucan, in oat products is an active hypolipidemic component that is responsible for lowering plasma lipids. Quantitative analysis of β‐glucan in oat hydrocolloids such as Oatrim, Nutrim, and C‐Trim was performed to measure the total β‐glucan content and molecular weight distribution. For the measurement of total β‐glucan content, both modified flow‐injection analysis (FIA) method and the standard AACC enzymatic method were employed. FIA method uses the enhanced fluorescence produced when β‐glucan forms complexes with Calcofluor. Experimental results of both the modified FIA method and the standard AACC enzymatic method revealed very good coincidence with each other. This result confirms the applicability of either technique for the quantitative evaluation of β‐glucan in hydrocolloids. Molecular weight (MW) distribution of β‐glucan was determined by size‐exclusion chromatography with postcolumn detection. Experimental results revealed that the molecular weight of β‐glucan in the Trim products was decreased during the manufacturing process. This result was ascribed to the rigorous processing condition of jet‐cooking.     

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.     

Glycemic Response to Oat Bran Muffins Treated to Vary Molecular Weight of β‐Glucan

Oat bran muffins, containing 4 or 8 g of β‐glucan per two‐muffin serving, were prepared with or without β‐glucanase treatment to produce a range of β‐glucan molecular weights from 130,000 to just over 2 million. Following an overnight fast, the glycemic responses elicited by the untreated and treated muffins was measured in 10 healthy subjects and compared with a control whole wheat muffin. Taken all together, the 4‐g β‐glucan/serving muffins reduced blood glucose peak rise (PBGR) by 15 ± 6% compared with the control. The 8‐g β‐glucan/serving muffins had a significantly greater effect (44 ± 5% reduction compared with the control, P < 0.05). The efficacy of the muffins decreased as the molecular weight was reduced from a 45 ± 6% reduction in PBGR (P < 0.05) for the untreated muffins (averaged of both serving sizes) to 15 ± 6% (P < 0.05) for muffins with the lowest molecular weight. As the molecular weight was reduced from 2,200,000 to 400,000, the solubility of the β‐glucan increased from a mean of 44 to 57%, but as the molecular weight was further decreased to 120,000, solubility fell to 26%. There was a significant correlation (r² = 0.729, P < 0.001) between the peak blood glucose and the product of the extractable β‐glucan content and the molecular weight of the β‐glucan extracted.     

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.     

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.     

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.     

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.     

Effect of Formulation and Processing Treatments on Viscosity and Solubility of Extractable Barley β‐Glucan in Bread Dough Evaluated Under In Vitro Conditions

β‐Glucan shows great potential for incorporation into bread due to its cholesterol lowering and blood glucose regulating effects, which are related to its viscosity. The effects of β‐glucan concentration, gluten addition, premixing, yeast addition, fermentation time, and inactivation of the flour enzymes on the viscosity of extractable β‐glucan following incorporation into a white bread dough were studied under physiological conditions, as well as, β‐glucan solubility in fermented and unfermented dough. β‐Glucan was extracted using an in vitro protocol designed to approximate human digestion and hot water extraction. The viscosity of extractable β‐glucan was not affected by gluten addition, the presence of yeast, or premixing. Fermentation produced lower (P ≤ 0.05) extract viscosity for the doughs with added β‐glucan, while inactivating the flour enzymes and increasing β‐glucan concentration in the absence of fermentation increased (P ≤ 0.05) viscosity. The physiological solubility of the β‐glucan concentrate (18.1%) and the β‐glucan in the unfermented dough (20.5%) were similar (P > 0.05), while fermentation substantially decreased (P ≤ 0.05) solubility to 8.7%, indicating that the reduction in viscosity due to fermentation may be highly dependent on solubility in addition to β‐glucan degradation. The results emphasize the importance of analyzing β‐glucan fortified foods under physiological conditions to identify the conditions in the dough system that decrease β‐glucan viscosity so that products with maximum functionality can be developed.     

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.     

Physical and Sensory Characteristics of Extruded Products Made from Two Oat Lines with Different β‐Glucan Concentrations

The impact of extrusion on physical and sensory properties and on the in vitro bile acid (BA) binding was examined for N979 and Jim oat (Avena sativa) lines with 8.1 and 4.8% β‐glucan, respectively. Based on hardness and edibility of products made from Jim oats, moisture concentrations of 16–25% and temperatures of 165–180°C were selected for N979 extrusion. Jim‐based cereal had a significantly greater (P < 0.05) expansion ratio than did N979‐based cereal at most moistures. N979 cereal was browner, but not harder, than Jim cereal. Extruded products from N979 and Jim oats had 5.29–5.99% and 3.38–3.94% β‐glucan, respectively. Changing extrusion temperature or moisture content did not affect β‐glucan concentration in the products. N979 cereal made at 165°C and 16% moisture had greater BA binding than at other conditions, and had crunchiness comparable to cereals made at other conditions. BA binding of Cheerios brand breakfast cereal was close to that of N979 cereal made at 180°C and 18% moisture, but lower than cereals made at other conditions. Cereals made from Jim and N979 oats were browner, harder, coarser, and crunchier than Cheerios breakfast cereal. Proper processing and preparation techniques should be considered when producing extruded products from high β‐glucan oats.     

Development of a Germination Process for Producing High β‐Glucan,Whole Grain Food Ingredients from Oat

Germination can be used to improve the texture and flavor of cereals. However, germination generally causes breakdown of β‐glucans, which is undesirable with respect to the functional properties of β‐glucan. Our aim was to assess possibilities of germinating oat without substantial loss of high molecular weight β‐glucan. Two cultivars, hulled Veli and hull‐less (naked) Lisbeth were germinated at 5, 15, and 25°C and dried by lyophilization or oven drying. Elevated germination temperatures led to an increase in Fusarium, aerobic heterotrophic bacteria, Pseudomonas spp., lactic acid bacteria, enterobacteria, and aerobic spore‐forming bacteria. Therefore, the germination temperature should be kept low to avoid excessive growth of microbes. Of the samples germinated at 15°C, only one contained low amounts of the Fusarium toxin deoxynivalenol (52 μg/kg). Germination led to the breakdown of β‐glucans, but the decrease in the molecular weight of β‐glucan was initially very slow. A short germination schedule (72 hr, 15°C) terminated with oven drying was developed to produce germinated oat with retained β‐glucan content. Compared with the native oat, 55–60% of the β‐glucan could be retained.     

Extraction of β‐Glucan from Oat Bran in Laboratory Scale

Effects of various enzymes and extraction conditions on yield and molecular weight of β‐glucans extracted from two batches of commercial oat bran produced in Sweden are reported. Hot‐water extraction with a thermostable α‐amylase resulted in an extraction yield of ≈76% of the β‐glucans, while the high peak molecular weight was maintained (1.6 × 10⁶). A subsequent protein hydrolysis significantly reduced the peak molecular weight of β‐glucans (by pancreatin to 908 × 10³ and by papain to 56 × 10³). These results suggest that the protein hydrolyzing enzymes may not be pure enough for purifying β‐glucans. The isolation scheme consisted of removal of lipids with ethanol extraction, enzymatic digestion of starch with α‐amylase, enzymatic digestion of protein using protease, centrifugation to remove insoluble material, removal of low molecular weight components using dialysis, precipitation of β‐glucans with ethanol, and air‐drying.