Relationship between specific weight of spring barley and malt quality

The assessment of malting barley to determine if it meets grain quality requirements is an integral step in ensuring an efficient malting process and a good quality malt output. Specific weight (SW) is an industry standard criterion, however links between SW and malting are not well understood. In this study the effect of a changing SW on malting was investigated. Samples were manipulated according to both grain size and weight, creating grain fractions with a range in SW. Prior to malting, grain quality traits were measured, and after malting, malt quality traits were examined. Increased SW resulted in a reduced number of whole, unmodified corns in malt, implying increased levels of modification. Specific weight correlated with both hot water malt extract (r = 0.82, P < 0.01) and predicted spirit yield (r = 0.84, P < 0.01), this highlights an increased malt output. Furthermore peak gelatinisation temperature of extracted starch from the malt correlated with both SW (r = 0.69, P < 0.05) and grain density (r = 0.65, P < 0.05). This could benefit malt efficiency by increased conversion of starch to fermentable sugars, but with the same energy input. The changes in SW and consequently malt output in this study are a result of changing grain density rather than packing efficiency.     

Combining ability of barley flour pasting properties

Malt extract is one of the major traits contributing to high malting quality and thus a major objective in malting barley breeding programs. Understanding the genetic behaviour of this trait could make selection more efficient. However, the measurement of this trait is very time-consuming and cannot be done in a single plant since it needs a certain amount of grain. In this paper, 10 different varieties with different malting quality were selected to make two 6 × 6 half-diallel crosses for the purpose of studying the genetic behaviour of flour pasting properties, in particular pasting temperature which has been shown to be closely related with malt extract. The pasting properties were measured with a Rapid ViscoAnalyser. Both Australian malting varieties showed significantly higher general combining ability for lower pasting temperature (higher malt extract). The Japanese malting variety also appeared to be a good parent for lower pasting temperature. All the other feed varieties which are used in the breeding programs for improving waterlogging tolerance or salinity tolerance showed significantly higher pasting temperature. Since specific combining ability was not significant, the selection of parents when breeding for pasting temperature can be based on the pasting temperature of the parents. Combined with the fact that only a small amount of grain is needed for the measurement of pasting temperature, the selection can be made in early segregating generations. Preliminary study showed that a major QTL associated with pasting temperature was located on chromosome 7H, indicating the possibility of marker assisted selection for this trait.     

Combining ability of barley flour pasting properties

Malt extract is one of the major traits contributing to high malting quality and thus a major objective in malting barley breeding programs. Understanding the genetic behaviour of this trait could make selection more efficient. However, the measurement of this trait is very time-consuming and cannot be done in a single plant since it needs a certain amount of grain. In this paper, 10 different varieties with different malting quality were selected to make two 6 × 6 half-diallel crosses for the purpose of studying the genetic behaviour of flour pasting properties, in particular pasting temperature which has been shown to be closely related with malt extract. The pasting properties were measured with a Rapid ViscoAnalyser. Both Australian malting varieties showed significantly higher general combining ability for lower pasting temperature (higher malt extract). The Japanese malting variety also appeared to be a good parent for lower pasting temperature. All the other feed varieties which are used in the breeding programs for improving waterlogging tolerance or salinity tolerance showed significantly higher pasting temperature. Since specific combining ability was not significant, the selection of parents when breeding for pasting temperature can be based on the pasting temperature of the parents. Combined with the fact that only a small amount of grain is needed for the measurement of pasting temperature, the selection can be made in early segregating generations. Preliminary study showed that a major QTL associated with pasting temperature was located on chromosome 7H, indicating the possibility of marker assisted selection for this trait.     

Volatile metabolite profiling of malt contaminated by Fusarium poae during malting

A volatile metabolite-profiling approach based on solid-phase microextraction (SPME) combined with gas chromatography–mass spectrometry was used to investigate volatile metabolic changes over the course of a malting process contaminated by Fusarium poae to assess its influence on malt flavor. GC–MS analysis of time-serial samples revealed a broad spectrum of volatile compounds including aldehydes, ketones, alcohols, organic acids, aromatics and furans. Statistical assessment of the data via principal component analysis demonstrated that at the initial phase of germination, the influence of F. poae was relatively small. However, its influence increased over time, resulting in differences between contaminated and controlled samples at 120 h and 144 h. Analysis of the corresponding loadings showed that volatile metabolites from green malt were influenced by contamination with F. poae fungi during malting to an extent, especially for the volatile aldehyde fractions. These compounds can be formed during both primary and secondary metabolism, and the potential exists for metabolic perturbation resulting from F. poae.     

Effect of hydrogen peroxide and ozone treatment on improving the malting quality

The effect of hydrogen peroxide (HP) and ozone (O₃) treatment during barley steeping on the quality of malt produced from two barley varieties (GrangeR and AC Metcalfe) by micro-malting was investigated. The two steeping oxidation treatments that was observed to promote barley acrospire growth. Ozone treatment improved the malt enzyme activity of endo-protease, α-amylase, free beta-amylase and total limit dextrinase to differing extents, with GrangeR improving to a greater degree. HP treatment contributed to the increase of α-amylase, β-glucanase and endo-protease. Surprisingly, HP or ozone oxidation during malting resulted in different and novel outcomes for total beta-amylase in GrangeR and AC Metcalfe. In GrangeR, total beta-amylase activity reduced with respect to the control in both treatments. In comparison with AC Metcalfe there was a substantial increase of 78% with HP and 90% O₃ in total beta-amylase activity. Malt quality including wort free amino nitrogen, β-glucan, turbidity and diastatic power was differentially increased by the oxidation induction treatment during steeping in malting. Gene expression analysis indicated that the effects of the steep oxidation treatments on enzyme and malt quality were putatively linked with the up-regulation of certain genes involved in GA synthesis (GA20ox1) and ABA catabolism (ABA8′OH). Barley grain germination assay results also showed that moderate HP induction could improve barley germination tolerance to the ABA effect. Malting including steep oxidation induction was shown to be beneficial to malt quality by improving the resultant wort quality and the efficiency of the beer brewing process. These observations point the way towards improving malt quality and the efficiency of the malting process.     

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.     

Influence of germination time and temperature on the properties of rye malt and rye malt based worts

The effects of germination time and temperature on the quality of rye malt and worts derived thereof were investigated using Response Surface Methodology. Amylolytic and proteolytic enzyme activities were increased by long germination periods, while β-glucanase activity was not influenced. Total and Soluble Nitrogen content were also not significantly affected by the variations in germination conditions. Free Amino Nitrogen (FAN) was found in higher amounts in worts prepared from rye malts with long germination times. Extract contents were higher in rye malt than in the control barley malt and could be increased by a favourable germination regime, while no such impact on wort fermentability was found. High wort viscosities could be significantly reduced by a long germination period at low temperatures, but were still unacceptably high. The same conditions favoured the development of endoxylanase activity. Arabinoxylan (AX) accumulated during the germination process and their extractability increased. The results suggest that longer germination periods resulted in an increased number of AX molecules with lower molecular mass. Optimal rye malt qualities within the limits of this study were found for a germination time of 144 h at 10 °C, which resulted in an acceptable FAN content and the lowest measured viscosity.     

Predicting pre-harvest sprouting susceptibility in barley: Looking for “sensitivity windows” to temperature throughout grain filling in various commercial cultivars

Pre-harvest sprouting (PHS) is common in cereals that lack grain dormancy if maturing grain is exposed to rain. This phenomenon leads to immediate loss of seed viability, and since the malting process requires germination, its occurrence is highly undesirable in malting barley crops. Dormancy release rate is genetically and environmentally controlled. We evaluated the effect of temperature during grain filling on the dormancy release pattern (and then on the PHS susceptibility) of grains from five malting barley (Hordeum vulgare L.) cultivars widely sown in Argentina, with the aim of predicting PHS susceptibility of a barley crop from easy-to-gather data. Barley cultivars (Quilmes Ayelén, Q. Palomar, Q. Painé, B1215 and Scarlett) were sown on different dates over a 3-year period for generating variability in the thermal environment during grain filling. The period from pollination to physiological maturity (PM) was adjusted to a thermal time (TT) scale, which was then arbitrarily divided into 50 °C d intervals. Mean air temperature within each interval and for the whole filling period was calculated for the different sowing dates. Dormancy release pattern was followed by determining a weighed germination index (GI) throughout grain filling and maturation. We sought a linear relationship between temperature during grain filling and GI at some moment after PM. For all barley cultivars, except B1215, a significant (p < 0.001) and positive correlation was found between the GI of grains with 10–20% moisture content (fresh basis) and mean temperature within TT intervals located at the last stages of seed development. Then, simply temperature-based models for predicting crop PHS susceptibility were generated for each barley cultivar. Moreover, we intended a single, universal prediction model constructed with data from all cultivars. Two general forms were proposed, but the relationships were slightly less tight when each barley cultivar model was used. A preliminary validation for each cultivar model was done for three genotypes with independent data from four sites of the major barley production area in Argentina. When comparing experimental and field data regressions we did not find significant differences in slope for any cultivar (p > 0.25). However, most of the observed GIs were higher than predicted. This upwards displacement of GI–temperature relationship suggests the role of other environmental factors (i.e. water and soil N availability, day length), differing among tested locations. We are currently evaluating and quantifying the effect of these factors with the aim of improving PHS susceptibility prediction in malting barley crops.     

Effect of the use of dilute alkaline prior to Bacillus subtilis-based biocontrol steeping and germination conditions on red sorghum malt β-glucanase activities and residual β-glucans

Malting is the ideal stage to deal with β-glucans. Their hydrolysis is very important as the diffusion of both hormones and hydrolytic enzymes in the endosperm of germinated grain depend on it. A high malt β-glucanase activity is not a guarantee of an extensive hydrolysis of β-glucans. When Bacillus subtilis is used to control mould growth, red sorghum malt β-glucanase activity (measured using carboxymethylcellulose as the substrate) was improved without significantly affecting the hydrolysis of malt β-glucans. Thus, in order to reduce the residual β-glucans content, soaking in 0.2% NaOH was combined with a biocontrol. Soaking in 0.2% NaOH is recognized as capable of improving grain hydration by opening-up the endosperm cell walls. The combined use of 0.2% NaOH with B. subtilis-based biocontrol treatments during red sorghum malting, leads to malt with increased β-glucanase activity and a significant reduction of residual β-glucans when compared with the 16 h biocontrol steeping without prior steeping in 0.2% NaOH. β-glucanase activity increases with increased germination temperature and time while, conversely, the residual β-glucans content of the malts decreases. Indeed, while the level of β-glucanase was not vastly different between the malts obtained after steeping in distilled water and those obtained after 8 h steeping in 0.2% NaOH followed by 8 h resteeping in distilled water (NaOH + H₂O treatment), their residual β-glucans levels differ significantly. B. subtilis-based treatment leads to malt with improved β-(1-3)- and β-(1-4)-glucanase activities without significantly improved malt β-(1-3),(1-4)-glucanase activity. While malts obtained after 84 h germination weren't significantly different in terms of malt β-(1-3),(1-4)-glucanase activities for all steeping treatments, the use of 0.2% NaOH steeping prior to resteeping led to malts with improved β-glucans content. Combining the steeping in dilute alkaline and biocontrol enables taking advantage of the dilute alkaline effect on residual β-glucans content, due probably to the opening-up of the cell walls and the improvement of water uptake, and that of the biocontrol (improvement of β-glucanase synthesis).     

The Relationship Between D Hordein and Malting Quality in Barley

The contribution of grain protein to the malting quality of barley (Hordeum vulgareL.) was investigated by comparing the hordein composition and the malting quality in barley produced under a range of field conditions. Two malting cultivars, Schooner and Arapiles, and one feed cultivar, Galleon, were grown under five nitrogen regimes in each of two years. Hordein composition of the grain was determined at maturity using a combination of sodium dodecyl sulphate-polacrylamide gel electrophoresis and laser densitometry. Malt extract was determined on all samples after micromalting. Variation in growth conditions resulted in a wide range of grain protein contents and malt extract values, as well as variation in the proportions of the individual B, C and D hordeins in the grain. D hordein in particular varied over a 10-fold range. High levels of all protein fractions were associated with low malt extract. Total protein, as expected, displayed a strong, negative correlation with malt extract. The relationship was cultivar specific and separate regression lines were generated for each cultivar. Of the individual hordein fractions, D hordein displayed the strongest negative correlation with malt extract and its relationship to malt extract was independent of cultivar. A consistent relationship between D hordein and malt extract was observed across seasons, treatments and cultivars that was indicative of a causal relationship between D hordein and malting quality. D hordein therefore offers an alternative measurement to total protein for the prediction of malting quality over a wide range of environmental conditions and cultivars.     

Ukrainian Dietary Bread with Selenium-Enriched Soya Malt

The production of bread with addition of selenium-enriched soya malt was studied. Processing of this soya malt included soaking of the soya beans in the solution of hydroselenite with concentration 1.5 mg Se/L (20 μg of Se per 1 g of soya beans), then 4 days of beans germination at 20 °C, drying at 50 °C until moisture content 8%, separation from the sprouts and grinding. The soya malt was a powder containing 15–18 μg of Se in 1 g. The accumulated selenium was mainly in the protein fraction of soya malt. Addition of selenium-enriched soya malt to leaven intensified activity of yeasts and lactic acid bacteria. The quality of the wheat bread with selenium-enriched soya malt was better than that of the bread in control. The enriched bread had specific pleasant smell and soft texture. The daily intake of 277 g of bread with the selenium-enriched soya malt, which is added in quantity of 1.0–1.75% to mass of plain flour, ensures the consumption of 30–50% of selenium recommended daily allowance for 17 million population of the northern and northwestern Ukraine.

     

Amplified Fragment Length Polymorphism (AFLP) Markers for Barley Malt Fingerprinting

Barley is the cerealpar excellencein the production of malt for beer and other alcoholic beverages. In this work, biochemical (hordein A-PAGE) and molecular (RAPDs, AFLPs) markers have been compared for their efficiency in malt fingerprinting. To overcome the difficulties related to the quality of the DNA extracted from grain malt, malted coleoptiles are proposed as useful tissue for the extraction. On the basis of our data, we exclude a fingerprinting system based on A-PAGE hordein analysis because of the degradation of these proteins during the malting process. Comparing molecular markers for their efficiency and repeatability, we indicate AFLP analysis based on template DNA extracted from the tissue of grain malt coleoptile as an excellent tool for malt fingerprinting.     

Digestion of barley malt porridges in a gastrointestinal model: Iron dialysability,iron uptake by Caco-2 cells and degradation of β-glucan

Iron availability and degradation of (1→3,1→4)-β-d-glucan (β-glucan) in three whole grain porridges made from two optimised barley malts and unprocessed barley were studied in a dynamic gastrointestinal model. The malting processes, with steeping at 15 or 48 °C with lactic acid (LA), enabled a complete reduction of phytate by subsequent soaking of ground malt, still with well preserved β-glucan. Iron dialysability and iron uptake by Caco-2 cells were higher in phytate reduced porridges, compared to the reference porridge. During simulated digestion, the extractability of β-glucan increased and the Calcofluor average molecular weight decreased for all porridges, indicating a gradual degradation during passage through the model. The degradation rate, however, appeared lower in porridge prepared from malted barley steeped at 48 °C with LA. The gastrointestinal model ranked iron availability according to human absorption data and showed high repeatability when evaluating changes in β-glucan. The results indicate the potential for using high temperature steeping with LA to yield improved iron availability combined with reduced degradation of β-glucan in the small intestine, maintaining the beneficial properties of barley.     

The endogenous endoprotease inhibitors of barley and malt and their roles in malting and brewing

Endoproteases play an important role in barley germination by controlling the hydrolysis of the grain's storage proteins into peptides and amino acids that are needed by the young plant. During malting, the commercial version of this process, many high M_r barley biopolymers are converted into malt nutrients that can be utilized by yeasts during brewing. However, barley and malt both contain endogenous proteins that inhibit the enzymatic activities of these proteases. High levels of these inhibitors can cause brewing problems by preventing the proteases from producing optimal levels of soluble proteins and amino acids. Both high and low M_r inhibitors of cysteine proteases occur in barley and malt. Two of the high M_r inhibitors, lipid transfer protein 1 (LTP1) and LTP2, have been purified and studied. Recently, members of the trypsin/alpha-amylase inhibitor protein family (CM proteins) have been shown to inhibit the activity of SEP-1, a purified serine class barley protease. No inhibitors of aspartic proteases or metalloproteases have yet been purified, but it has been reported that endogenous metalloprotease inhibitors do exist. The inhibitors of the cysteine proteases and metalloproteases are probably the ones most important for brewing, because members of these two protease classes apparently catalyse most of the protein hydrolysis that occurs during malt mashing and, presumably, also during malting. More biochemical studies are needed to clarify how these proteins interact with the proteases to control protein hydrolysis during germination.     

The endogenous endoprotease inhibitors of barley and malt and their roles in malting and brewing

Endoproteases play an important role in barley germination by controlling the hydrolysis of the grain's storage proteins into peptides and amino acids that are needed by the young plant. During malting, the commercial version of this process, many high M_r barley biopolymers are converted into malt nutrients that can be utilized by yeasts during brewing. However, barley and malt both contain endogenous proteins that inhibit the enzymatic activities of these proteases. High levels of these inhibitors can cause brewing problems by preventing the proteases from producing optimal levels of soluble proteins and amino acids. Both high and low M_r inhibitors of cysteine proteases occur in barley and malt. Two of the high M_r inhibitors, lipid transfer protein 1 (LTP1) and LTP2, have been purified and studied. Recently, members of the trypsin/alpha-amylase inhibitor protein family (CM proteins) have been shown to inhibit the activity of SEP-1, a purified serine class barley protease. No inhibitors of aspartic proteases or metalloproteases have yet been purified, but it has been reported that endogenous metalloprotease inhibitors do exist. The inhibitors of the cysteine proteases and metalloproteases are probably the ones most important for brewing, because members of these two protease classes apparently catalyse most of the protein hydrolysis that occurs during malt mashing and, presumably, also during malting. More biochemical studies are needed to clarify how these proteins interact with the proteases to control protein hydrolysis during germination.     

Influence of high temperature and terminal moisture stress on dormancy in wheat (Triticum aestivum L.)

Preharvest sprouting is common in cereals that lack grain dormancy if maturing grain is exposed to rain. Over three successive seasons wheat genotypes with a range of dormancy levels, were exposed to moisture stress and periods of high temperature stress (>30 °C) in controlled field trials. Dormancy assessments were based on a germination index of hand threshed grain throughout grain filling. There were three main results. First, moisture stress combined with consistently high temperature during grain filling was associated with induced dormancy in Cunderdin, (germination index of 0.41) in a normally non-dormant genotype (germination index normally >0.80), but no additional dormancy in DM 2001, a dormant genotype (germination index normally <0.10). In contrast sudden heat shocks (>30 °C max. for >12 days) at 30–50 days post-anthesis reduced dormancy, germination index increase of 0.42 on average across five genotypes. Secondly, whilst dormancy was affected by moisture and heat stress, genotypes maintained their relative rankings across environments and genotype had the most effect on dormancy (70–92% of the variation in germination index) with DM 2001 and DH 22 more dormant than DH 56, DH 45 and Cunderdin. Finally, the effect of environment was different for different genotypes; those with partial dormancy (germination index usually 0.20–0.50, DH 56 and DH 45) were most influenced by the environmental conditions with germination indexes ranging from 0.06 to 0.85 depending on environment. Consequently avoidance of high temperatures, moisture stress, and maturity × stress interactions, are important prerequisites in screening for genotypes with genetic differences in dormancy.     

Malt hydrolysates for gluten-free applications: Autolytic and proline endopeptidase assisted removal of prolamins from wheat,barley and rye

Cereal based products intended for gluten sensitive individuals, particularly to celiac disease patients, tend to have poor organoleptic qualities and they contain low levels of healthy whole grain compounds. Adding whole grain ingredients, such as malt hydrolysates, could compensate these defects provided that the ingredients are adequately free from toxic prolamin epitopes. Here we demonstrate that the level of toxic prolamin epitopes in the malt autolysates (wheat, barley, rye) were substantially lower than in the native malts but too high to allow “very low in gluten” labelling. To further eliminate the residual levels of toxic prolamin epitopes, a proline-specific endoprotease from Aspergillus niger was added to the malt autolysates. In the resulting malt hydrolysates (of wheat and rye but not barley), the prolamins were indeed greatly reduced and were below the very low gluten limit of 100 mg/kg. Malt hydrolysates with adequately low gluten levels may potentially be used as novel ingredients within gluten-free foods.     

Electrophoretic analysis of malting degradability of major sorghum reserve proteins

Proteolysis is vital to the generation of amino acids and short peptides during malting. The qualitative and quantitative effects of malting on proteolytic digestion were investigated for 11 Botswana sorghum cultivars. Protein hydrolysis was influenced by sorghum grain cultivar. All protein fractions were degraded, although the extents of their digestion appeared cultivar-dependent. The most significant changes in total and free kafirins occurred among the HMW aggregates while the 45 kDa dimer appeared the most recalcitrant. Free kafirin monomers were digested in variety-defined manner; α-kafirin faded the most (ca. 82%) in Phofu, but appeared least digested (≤10%) in Lekgeberwa; β-kafirin faded the most in Phofu and Segaolane (90–94%) but only 46% in Lekgeberwa. Overall, α-kafirin appeared the least digested of the free monomers. The proportion of free kafirin accounted for by the 45 kDa and monomeric kafirins increased in all malts (except Lars Vyt and Mafia), probably due to protein depolymerisation. Grain variety significantly influenced (p < 0.001) malt FAN levels. Mean FAN values were highest in Segaolane (225 mg/100 g) followed by Sefofu, Town and BSH-1, but lowest in Mafia (96.9 mg/100 g). Results indicate wide variability in the proteolytic malting digestibility of the eleven sorghum cultivars.