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.     

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.     

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.     

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.     

Resistant Starch and Total Dietary Fiber Content of Oatrim Muffins with Different Levels of Amylose,Amylopectin, and β‐Glucan

Nine types of muffins made with three levels of β‐glucan and three amylose‐amylopectin ratios were prepared at the Beltsville Human Nutrition Research Center, United States Department of Agriculture. They were fed to human subjects to study effects of starch composition and dietary fiber content on the carbohydrate and lipid metabolism in normal and overweight women. The main objective of this study was to determine resistant starch (RS) and total dietary fiber (TDF) content of the muffins 1) using AACC Approved Method 32‐07 and AOAC method 991.43, incorporating a pretreatment step with dimethyl sulfoxide (DMSO) before enzyme incubation, 2) with pretreatment at 100 and 121°C before incubation with amyloglucosidase, and 3) using samples chewed by human subjects before incubation with pancreatin and amyloglucosidase. For method 1, on an as‐eaten basis, TDF content was 2.81 to 9.64 g/100 g for samples without DMSO pretreatment and 1.66 to 4.06 g/100 g with DMSO pretreatment. RS content was 0.30 to 11.18 g/100 g for methods 1 and 2, respectively. Methods 2 and 3 had the best correlation for all muffins tested (r² = 0.97).     

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.     

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.     

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.     

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.     

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.     

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.     

Textural and Bile Acid‐Binding Properties of Muffins Impacted by Oat β‐Glucan with Different Molecular Weights

Water‐soluble β‐glucan (BG) extracted from a high‐BG oat line was treated with different amounts of lichenase (1→3)(1→4)‐β‐d ‐glucanase) enzyme to yield three different molecular weight (MW) BG extracts. Low (LMW‐BG, 157,000), medium (MMW‐BG, 277,000), and high (HMW‐BG, 560,000) MW BG extracts were added to plain muffin formulations at a level of 0.52% (0.42% in the batter, 0.52% in the resultant muffins) to investigate the effect of MW of BG on textural and bile acid (BA) binding properties of the muffins. In addition, treatments were prepared containing LMW‐BG, MMW‐BG, and HMW‐BG extracts in amounts providing equivalent batter firmness as determined on a texture analyzer. Resultant BG concentrations (and per serving amounts) of these muffins were 1.36% (0.81 g/60 g muffin), 1.05% (0.63 g/60 g muffin), and 0.52% (0.31 g/60 g muffin), respectively; thus, the LMW treatment complied with a U.S. Food and Drug Administration health claim requiring 0.75 g of BG per serving. The firmness, springiness, and BA‐binding capacity of the muffins were unaffected by the MW of BG. However, when added at the maximum limit for equivalent batter firmness, the LMW treatment was more firm and less springy than the HMW treatment. Furthermore, BA‐binding capacities of LMW and MMW fractions tended to be greater than that of the HMW fraction when added at the maximum limit. These results add further evidence to the importance of fine‐tuning BG structure to provide maximum health benefits while maintaining high product quality.     

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.     

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.     

Fermentability of Oat and Wheat Fractions Enriched in β‐Glucan Using Human Fecal Inoculation

Fermentation by human fecal bacteria of fractions of wheat bran prepared by preprocessing technology were examined and compared with a β‐glucan‐rich oat bran and a purified β‐glucan (OG). The wheat fractions were essentially a beeswing bran (WBA), mainly insoluble dietary fiber, and an aleurone‐rich fraction (WBB) containing more soluble fiber and some β‐glucan (2.7%). The oat bran (OB) had more endosperm and was very rich in β‐glucan (21.8%). Predigestion of WBB and OB to mimic the upper gastrointestinal (GI) tract gave digested wheat bran fraction B (WBBD) and digested oat bran (OBD), respectively. These predigested fractions were fermented in a batch technique using fresh human feces under anaerobic conditions. Changes in pH, total gas and hydrogen production, short chain fatty acids (SCFA), and both soluble and insoluble β‐glucan and other polysaccharide components, as determined from analysis of monosaccharide residues, were monitored. Fractions showed increasing fermentation in the order WBA < WBBD < OBD < OG. Variations in SCFA production indicated that microbial growth and metabolism were different for each substrate. Polysaccharide present in the supernatant of the digests had disappeared after 4 hr of fermentation. Fermentability of oat and wheat β‐glucan reflected solubility differences, and both sources of β‐glucan were completely fermented in 24 hr. Although the overall patterns of fermentation indicated the relative amounts of soluble and insoluble fiber, the anatomical origin of the tissues played a major role, presumably related to the degree of lignification and other association with noncarbohydrate components.     

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.     

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.     

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.     

Microenzymatic Evaluation of Oat (Avena sativa L.) β‐Glucan for High‐Throughput Phenotyping

Oats (Avena sativa L.) have received significant attention for their positive and consistent health benefits when consumed as a whole grain food, attributed in part to mixed‐linkage (1‐3,1‐4)‐β‐d ‐glucan (referred to as β‐glucan). Unfortunately, the standard enzymatic method of measurement for oat β‐glucan is costly and does not provide the high‐throughput capability needed for plant breeding in which thousands of samples are measured over a short period of time. The objective of this research was to test a microenzymatic approach for high‐throughput phenotyping of oat β‐glucan. Fifty North American elite lines were chosen to span the range of possible values encountered in elite oats. Pearson and Spearman correlations (r) ranged from 0.81 to 0.86 between the two methods. Although the microenzymatic method did contain bias compared with the results for the standard streamlined method, this bias did not substantially decrease its ability to determine β‐glucan content. In addition to a substantial decrease in cost, the microenzymatic approach took as little as 6% of the time compared with the streamlined method. Therefore, the microenzymatic method for β‐glucan evaluation is an alternative method that can enhance high‐throughput phenotyping in oat breeding programs.