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

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

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.     

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.     

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.     

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.     

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.     

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.     

Waxy Starch Barley Genotypes with Reduced β-Glucan Contents

Random inbred lines were produced from a cross between the genotypes Chalky Glenn and Waxy Hector, and two-row lines were classified as waxy or nonwaxy by an iodine staining test. Mean nitrogen and β-glucan contents of the waxy types were higher than those of the nonwaxy types but, in contrast to previous data, mean milling energies of the two groups were not significantly different. Waxy lines with low milling energy had much lower β-glucan levels than those with high milling energy, and they also demonstrated much more extensive cell wall modification during malting. From a trial grown the following season, the waxy types with low milling energy were again identified and had levels of β-glucan content similar to those of nonwaxy types. β-Glucan contents and, particularly, milling energies showed good agreement between seasons. It is suggested that, although waxy starch is usually associated with high β-glucan content, a genetic factor from Chalky Glenn that confers low levels of β-glucan can express in a waxy background.     

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.     

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.     

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.     

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

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

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.     

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.     

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.     

α-Amylolysis of Large Barley Starch Granules

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

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).     

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.     

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.