Cereal β-glucans in diet and health

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

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

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

Similarities and differences in secondary structure of viscoelastic polymers of maize α-zein and wheat gluten proteins

The secondary structure of a dough-like zein polymer was compared to the structure present in a wheat viscoelastic system using FT-IR spectroscopy. When zein was mixed at 35 °C, which is above its glass transition temperature (T_g), changes in its secondary structure suggested that the protein loses its native structure, mainly composed of α-helices (68%), and a viscoelastic system is formed by a structural rearrangement that favors β-sheet structures. This rearrangement is very similar to the structural changes observed in gluten viscoelastic polymers. Upon removal of shear stress, the zein polymer showed a rapid decrease in the proportion of β-sheet structures (from 48% to 28% after the first 3 min) in favor of unordered structures. At the same time, the viscoelasticity of the polymer decreased rapidly. In contrast, gluten, in a similar viscoelastic system and held at the same temperature, showed a fairly constant high content of β-sheet structures (49%) coinciding with the slow relaxation time typical of gluten networks after the removal of shear. We speculate that the addition of a protein capable of causing extensive and stable β-sheet formation in the zein–starch viscoelastic polymer could increase the stability and relaxation time of the zein system and, thereby, create the possibility of a zein dough with similar functionality to a wheat viscoelastic system.     

Genetic markers for CslF6 gene associated with (1,3;1,4)-β-glucan concentration in barley grain

The amount of (1,3;1,4)-β-D glucan accumulated in barley (Hordeum vulgare L.) cell walls is an important consideration for grain end-use. One of the major genes responsible for (1,3;1,4)-β-glucan biosynthesis is HvCslF6, which was analyzed in this study to determine the allelic variation between low (1,3;1,4)-β-glucan (∼3.3%) cultivar CDC Bold and high β-glucan (∼5.2%) line TR251. The CDC Bold HvCslF6 allele showed 16 single nucleotide polymorphisms (SNPs) and two indels when genomic region downstream of the ATG start codon was compared to TR251 allele. Both indels added 16 nucleotides to HvCslF6 first intron of CDC Bold and a single SNP in the third exon altered alanine 590 codon in the CDC Bold sequence to a threonine codon in TR251 allele. Genetic markers were developed for five polymorphic sites and confirmed useful to select low and high β-glucan lines in a previously characterized CDC Bold/TR251 mapping population and a novel F₅ recombinant inbred line (RIL) population derived from a Merit/H93174006 (4.8 and 5.3% (1,3;1,4)-β-glucan) cross. An analysis of parental lines of six populations segregating for β-glucan concentration validated association between the TR251 HvCslF6 haplotype and high (1,3;1,4)-β-glucan concentration in populations showing a (1,3;1,4)-β-glucan quantitative trait locus (QTL) on chromosome 7H.     

The identification of a barley haze active protein that influences beer haze stability: Cloning and characterisation of the barley SE protein as a barley trypsin inhibitor of the chloroform/methanol type

Previous pilot brewing trials have demonstrated that in the absence of a molecular weight (MW) 12,000 (barley silica eluate (SE) protein), the beer brewed from the malt of these SE −ve varieties formed less haze after accelerated ageing than beers brewed using SE +ve malt varieties. The previously described SE protein was characterised using comparative two-dimensional (2-D) gel electrophoresis immunoblots of barley seed extracts from both SE +ve and SE −ve varieties. The SE protein spot identified was excised and its partial sequence determined, after in-gel cleavage using trypsin and separation of the resulting fragments by reversed-phase high performance liquid chromatography (HPLC). N-terminal sequence analysis of the tryptic peptides from SE +ve and SE −ve varieties identified the SE protein as the barley trypsin inhibitor-CMe precursor (BTI-CMe). The mature BTI-CMe protein is 13.3 kDa and its functional gene is located on chromosome 3H. Cloning of the BTI-CMe protein demonstrated that both SE −ve and SE +ve barley varieties contain a BTI-CMe protein family member that is similar but has a consistently different sequence, primarily in the last 30 amino acid residues of their C-termini.     

The identification of a barley haze active protein that influences beer haze stability: The genetic basis of a barley malt haze active protein

In bright beers, the formation of haze is a serious quality problem, which places limitations on the storage life of the product. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) immunoblot analysis using an antiserum that was raised against a silica eluate (SE) protein fraction (obtained from silica gel, used for the colloidal stabilisation of beer), detected a range of protein bands in barley, malt, beer and haze. A polymorphism was observed in which some barley varieties contained a molecular weight (MW) 12,000 band (SE +ve) while in other varieties this band was absent (SE −ve). A survey of 219 Australian and international barley varieties, including a comprehensive selection of current and past malting varieties, identified 181 varieties as SE +ve, and 38 varieties as SE −ve. Previous pilot brewing trials demonstrated that SE −ve varieties are desirable as the beer brewed from the malt of these varieties formed less haze after accelerated ageing than beers brewed using SE +ve malt varieties. The genetic basis for the absence of the SE protein was conferred by a recessive allele at a single locus. Interval mapping analysis showed that the MW 12,000 band mapped to the short arm of chromosome 3H.     

Effect of particle size on kinetics of starch digestion in milled barley and sorghum grains by porcine alpha-amylase

The influence of milled grain particle size on the kinetics of enzymatic starch digestion was examined. Two types of cereals (barley and sorghum) were ground, and the resulting grounds separated by size using sieving, with sizes ranging from 0.1 to 3 mm. In vitro enzymatic digestion was performed, using pancreatic alpha-amylase, amyloglucosidase and protease, to determine fractional-digestion rates over 24 h. The resulting glucose production rate data were well fitted by simple first-order kinetics. For each sieve screen size, the digestion rate of barley was always higher than that of sorghum. The rate coefficients for digestion showed a decrease with increasing size, and could be well fitted by an inverse square relationship. This is consistent with the supposition that starch digestion in these systems is controlled by diffusion of enzyme through the grain fragment. Apparent diffusion coefficients of alpha-amylase obtained by fitting the size dependence were 0.76 (sorghum) and 1.7 (barley) × 10⁻⁷ cm² s⁻¹, 9 (sorghum) and 4 (barley) times slower than predicted for a molecule of the size of alpha-amylase in water.     

A chemometric evaluation of the underlying physical and chemical patterns that support near infrared spectroscopy of barley seeds as a tool for explorative classification of endosperm genes and gene combinations

Near infrared spectroscopic (NIR; 1100–2500 nm), chemical and genetic data were combined to study the pleiotropic secondary effects of mutant genes on milled samples in a barley seed model. NIR and chemical data were both effective in classifying gene and gene combinations by Principal Component Analysis (PCA). Risø mutants R-13, R-29 high (1→3, 1→4)-β-glucan, low starch and R-1508 (high lysine, reduced starch), near isogeneic controls and normal lines and recombinants were studied. Based on proteome analysis results, six anti-microbial proteins were followed during endosperm development revealing pleiotropic gene effects in expression timing that supporting the gene classification. To verify that NIR spectroscopy data represents a physio–chemical fingerprint of the barley seed, physical and chemical spectral components were partially separated by Multiple Scatter Correction and their genetic classification ability verified. Wavelength bands with known water binding and (1→3, 1→4)-β-glucan assignments were successfully predicted by partial least squares regression giving insight into how NIR-data works in classification. Highly reproducible gene-specific, covariate, pleiotropic classification patterns from NIR and chemical data were demonstrated in PCAs and by visual inspection of NIR spectra. Thus PCA classification of NIR-data gives the classical genetic concept, ‘pleiotropy’, a new operational definition as a fingerprint from a spectroscopic representation of the phenome carrying genetic, physical and chemical information. It is concluded that barley seed phenotyping by NIR and chemometrics is a new, reliable tool for characterising the pleiotropic effects of mutant gene combinations and other genotypes in selecting barley for quality in plant breeding.     

Effects of sulphur on yield and malting quality of barley

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

The effect of gluten on the retrogradation of wheat starch

The retrogradation of amylopectin in a wheat starch and a wheat starch/gluten (10:1) blend prepared by extrusion and containing 34% water (wet weight basis) was studied using X-ray diffraction, differential scanning calorimetry and NMR relaxometry during storage at constant water content and temperature (25 °C). For both samples, amylopectin ‘fully’ retrograded after 2–3 days storage, i.e. the different parameters monitored with time to follow the retrogradation had reached their maximum value, and crystallised predominantly into the A polymorph. Under the experimental conditions used, there was no evidence of any significant effects of the presence of gluten on the kinetics, extent or polymorphism of amylopectin retrogradation.     

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

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

Influence of growth temperature on content, viscosity and relative molecular weight of water-soluble β-glucans in barley (Hordeum vulgare L.)

β-Glucan content and viscosity of water-soluble β-glucans have a considerable impact on the digestion of barley. Eight different 2-row barley (Hordeum vulgare L.) cultivars were grown in controlled environment chambers at five different temperatures until maturity. The samples were pearled, milled and analysed for their content of total, insoluble and soluble β-glucan. The water-soluble fraction was extracted at 37 °C, subjected to amylase treatment and freeze-dried. Water-soluble crude β-glucans were solubilised in distilled water, analysed for viscosity and their purity was checked by nuclear magnetic resonance. Molecular weights were profiled by size exclusion chromatography with RI detection. The content of total β-glucan varied from 4.0% to 7.4%, and was significantly affected by the growth temperature. An interaction between cultivar and growth temperature was observed for the total β-glucan content. The extractability was significantly affected by growth temperature, as there was recorded an increasing amount of water-soluble β-glucan with increasing growth temperature. Both the viscosity and the molecular weight of the water-soluble β-glucans increased with the growth temperature.     

Molecular Basis of the Gelatinisation and Swelling Characteristics of Waxy Rice Starches Grown in the Same Location During the Same Season

Six waxy rice samples (grown at the same site during the same season) were investigated to identify those aspects of amylopectin structure and granule composition which differentiate the starches into high (T_p 77·8 °C on average) or low (T_p 66·2 °C on average) gelatinisation temperature. All starches contained less than 2·5% amylose with negligible lipid. Granule dimensions were similar (mean diameter 4·9–5·7 μm) as was the β-amylolysis limit (59·9–63·0%). However, the structural elements of amylopectins within the two groups varied. The low gelatinisation temperature starches contained two major chain fractions upon debranching with iso-amylase, F3 (DP 16) and F1 (DP 51) on average while the high gelatinisation temperature starches contained three fractions, F3 (DP 16), F2 (DP 19) and F1 (DP 40). The proportion of F1/(F2+F3) for the low and high gelatinisation starches representing on average 20·2 and 10·5% respectively. Hence, the higher proportion of exterior chains and especially those with DP 19 appear to confer the higher gelatinisation temperature characteristic of these starches. Debranched β-limit dextrins were prepared from these starches. Regardless of their origin, (high or low gelatinisation temperature), they contained three fractions: F_3β (DP 5), F_2β (DP 13) and F_1β (DP 19). Hence, the high versus low gelatinisation characteristic was imparted by the exterior chains. Using 13-C CP-MAS/NMR it was confirmed that the number of double helices within the starches were approximately constant although small differences in crystallinity were identified by X-ray diffraction. Retrogradation of the solubilised native high and low gelatinisation temperature amylopectin molecules confirmed that the ‘high temperature gelatinisation amylopectin’ molecules form higher dissociation temperature and enthalpy crystalline domains. This confirms that the higher proportion of short and especially relatively longer short chains promotes crystallite formation.     

Postharvest decay control and quality retention in litchi (cv. McLean's Red) by combined application of modified atmosphere packaging and antimicrobial agents

The effects of modified atmosphere packaging (MAP) [bioriented polypropylene (BOPP-1 or BOPP-2)] in combination with antimicrobial agents Bacillus subtilis, 10⁷ colony-forming units (cfu) ml⁻¹; ethylenediamine tetraacetic acid, calcium disodium salt hydrate (EDTA) (0.1%); or 4-hexylresorcinol (4-HR) (0.15%) on postharvest decay control and quality retention of litchi cv. McLean's Red were assessed as possible replacements for commercial SO₂ fumigation. Fruits dipped in B. subtilis, EDTA or 4-HR (5 min) separately, blow dried (25 °C, 3 min), packed in BOPP-1, held for 18 d at 2 °C, 95% RH, and 2 d at 14 °C, 75% RH were significantly less decayed. The antagonist–BOPP-1 combination also promoted the best bacterial survival during storage. B. subtilis was observed to survive effectively in BOPP-1 (16% O₂, 6% CO₂; 90% RH), but its survival was adversely affected in BOPP-2 (5% O₂, 8% CO₂; 93% RH). Alternaria alternata and Cladosporium spp. were the major decay-causing fungi in BOPP-1 treatments, and Candida, Cryptococcus and Zygosaccharomyces were the predominant yeasts in BOPP-2 treatments. Combination treatments EDTA, 4-HR or B. subtilis in BOPP-1 inhibited polyphenol oxidase (PPO) and significantly reduced pericarp browning and severity. Although the combination treatments EDTA, 4-HR or B. subtilis in BOPP were equally effective in controlling decay and browning, the EDTA and 4-HR affected the natural pinkish-red colour of the pericarp by showing higher h° values (orange–pink). Among the combination treatments, B. subtilis+BOPP-1 had the best potential to control decay, retain the colour and the overall litchi fruit quality during a marketing chain of 20 d.     

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

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

Barley genotypic β-glucan variation combined with enzymatic modifications direct its potential as a natural ingredient in a high fiber extract

β-Glucan dietary fiber rich food systems hold a great potential for improving human health. Here, we evaluate the extractability and quality of β-glucan liquid formulations from eight different barley genotypes varying in β-glucan content and molecular structure using Termamyl^® SC (T), Attenuzyme^® (A) and Attenuzyme^® Flex (AF) amylolytic enzymes in combinations. Extracts from barley lines Lys5f, KVL408, KVL1104 and CDC Fibar exceeded 4 g β-glucan/l, providing European Food Safety Authority (EFSA) and U.S. Food and Drug Administration (FDA) recommended amounts (3 g β-glucan/day) from three portions. TAF extracts of Lys5f and KVL408 grains reached extraordinary high concentrations of 8–9 g β-glucan/l. The β-glucan molecular mass decreased with enzyme treatment T < TA < TAF due to minor lichenase side activity. Extractability was generally higher and molecular mass lower for barley lines low in triosyl/tetraosyl (DP3/DP4) ratios than for those high in DP3/DP4 ratios (Lys5f, KVL408 and KVL1104). Overall, the higher β-glucan content and structural robustness in Lys5f and KVL408 raw materials favor these in a β-glucan rich extract with potential for EFSA and FDA health and nutrition claims.     

Relationship between the glass transition temperature and the melt flow behavior for gluten, casein and soya

The effects of moisture content (25–45% wwb) and temperature (75–120 °C) on the viscosity of gluten, soya and rennet casein systems was studied using a capillary rheometer. An attempt was made to relate the viscosities to the glass transition temperature measured by differential scanning calorimetry, dynamic mechanical thermal analysis and the phase transition analyzer. The temperature where the material flowed was also determined by the latter technique. All three-protein systems showed shear and extension thinning. Over the shear rate range investigated (1–10³ s⁻¹), gluten had a substantially lower viscosity than the other two proteins, although the difference was less pronounced at the highest temperature studied. This low viscosity is reflected by lower values of the glass transition temperature, the melt flow temperature and the dynamic moduli E′ and E″ in the rubbery state. The results are discussed in terms of the structure and heat induced changes for the three proteins and their relevance to food processing considered.     

Simulation of the factors affecting β-glucan levels during the cultivation of oats

A Monte Carlo simulation technique was employed to simulate the factors influencing the level of β-glucan content in both hulled (HO) and naked (NO) oat cultivars during cultivation of the crop. Probability density functions were used to model the uncertainty and variability in the input factors. A scenario analysis was subsequently developed to look at the impact of different model assumptions and input parameters. The simulated mean β-glucan level in harvested oats grain was 3.50 and 4.25 g/100 g for hulled (HO) and naked (NO), respectively. A sensitivity analysis highlighted that cultivar selection was the most important input parameter compared to other inputs in determining the final β-glucan level (correlation coefficients of 0.64 and 0.79 for HO and NO, respectively). The analysis also indicated the positive effect of delayed sowing on β-glucan content (correlation coefficients of 0.32 and 0.25 for HO and NO, respectively). Germination and storage factors showed a negative impact on the final β-glucan levels. The scenario analysis shows the applicability of the proposed model for various agronomic practices. This approach establishes a quantitative scientific ranking of factors influencing β-glucan levels during the cultivation of oats.     

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

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