Assessment of Enzymatic Endosperm Modification of Malting Barley Using Individual Grain Analyses

Enzymatic modification of the endosperm of malting barley is a main feature of the malting process. Uneven enzymatic modification of the endosperm (heterogeneity) can cause brewhouse problems. Although there is a general correlation between endosperm modification, beta‐glucan breakdown and endo‐beta‐glucanase development, it is based on average results from sample analyses and may conceal heterogeneity. The primary aim of this work was to use individual grain analyses to investigate factors that control endosperm modification and beta‐glucan breakdown. In terms of beta‐glucan breakdown and physical modification, the barley variety Chariot malted faster than Decanter. However, both varieties showed similar distribution of endo‐beta‐glucanase in individual grains during malting. Further work on individual grains showed that the correlation between beta‐glucan breakdown and endo‐beta‐glucanase activity was not significant. Surprisingly beta‐glucan breakdown did not always correlate with the physical modification of the endosperm. Both these findings suggest that sample analyses of beta‐glucan levels and malt beta‐glucanase activities are not reliable indicators of the degrees of which malt samples are modified during malting. Since the distribution of beta‐glucan in individual grains of the unmalted barley varieties was similar, the total beta‐glucan levels of the original barley did not determine the rate at which beta‐glucan was broken‐down during malting. Although protein studies are at a preliminary stage, the rate of protein breakdown was not correlated with the rate at which beta‐glucan was broken down in the malting grain. It is possible that the physico‐chemical properties of endosperm storage proteins may limit the rate of beta‐glucan breakdown during malting.     

Effects of Grain and Malt â‐Glucan on Distilling Quality in a Population of Hull‐less Barley

A population of barley lines, derived by mutation in the hull‐less variety, Penthouse, was included in a replicated trial, along with Penthouse and the hulled malting cultivar, Optic. Samples were assessed for a range of grain quality traits, then malted, with germination for either 4 or 5 days, prior to kilning. Most lines had grain β‐glucan contents lower than that of Penthouse, but there was no significant correlation between grain and malt β‐glucan content. Malt β‐glucan levels were indicative of differences in cell wall breakdown between 4 and 5 days germination, but negative associations with distilling parameters Extract and Alcohol Yield, were not statistically significant. It was concluded that the lines differed in the rate and extent of cell wall breakdown and that grain shape may influence modification in distal parts of the grain. However, a malting regime, optimised to suit Optic may be less suited to discriminating between hull‐less lines of reasonable quality.     

Malting Performance of Normal Huskless and Acid‐Dehusked Barley Samples

Sulphuric acid dehusked barley had a higher germinative energy and lower microbial infection than normal huskless (naked) barley, suggesting that the pericarp layer harboured microbial infection which may have limited the germination rate. Dehusking the normal huskless barley using sulphuric acid resulted in lower microbial infection, and increased germinative energy. The normal huskless barley sample had a higher β‐glucan content than the acid‐dehusked barley and had slower β‐glucan breakdown during malting. This resulted in the release of seven times more β‐glucan during mashing, and the production of wort of higher viscosity. The normal huskless barley sample had a higher total nitrogen content than the acid‐dehusked barley but both samples produced similar levels of amylolytic (α‐ and β‐amylase) activity over the same malting period. No direct correlation was found between barley total nitrogen level and the amylolytic activity of the malt. When barley loses its husk at harvest, the embryo is exposed and may be damaged. This may result in uneven germination which can reduce malting performance and hence malt quality.     

Malt Performance is More Related to Inhomogeneity of Protein and β‐Glucan Breakdown than to Standard Malt Analyses

Nitrogen analyses of the grains of samples of commercial malts indicate that β‐glucan breakdown and the uniformity of malt modification are influenced by uniformity of distribution of grain protein. It is proposed that for normal malting barley, variations in malt modification are related to the different percentages of grains which contain high levels and different types of proteins which resist enzymic modification during malting. This kind of inhomogeneity of malt modification can cause brewhouse problems but cannot be detected with precision by standard malt analyses.     

Barleys Grown as Cultivar Mixtures Compared with Blends Made Before and After Malting,for Effects on Malting Performance

Four barley cultivars were grown in replicated trials at three sites in Scotland in 2000, both as pure stands and in all four possible three‐component mixtures. After harvest, some grain from the pure stands was used to synthesise four blends of three component varieties. Grain from the pure stands, the mixtures and the blends was malted and all samples were assessed for total β‐glucan content. At two of the sites, field grown mixtures were shown to have lower malt β‐glucan than blends made prior to malting, although their grain β‐glucan contents had not been significantly different from the means of the component varieties. At the other site, the mixtures had higher levels of soluble nitrogen than the blends or the means of their component cultivars although, significant differences had not occurred in grain nitrogen contents. Three component blends were also made from the malted grain of the pure stands and hot water extracts were measured on all samples including the blends made before and after malting. There were considerable differences between sites and also between mixtures, blends and the mean of the mixture components when assessed separately. At all sites and for all varietal combinations, field grown mixtures were shown to be equal or superior to blends made after harvesting, in addition to frequently exceeding the mean of their components. It was concluded that the advantages, in β‐glucan or protein modification, associated with mixtures resulted from interactions between components in the growing environment and that interactions in the malting and mashing environments had little if any effect.     

The structure of barley endosperm – an important determinant of malt modification

Structural differences in barley grains have been classified as either mealy or steely and their relative proportions have been determined using a light transflectance method in three barley samples varying in the degree of steeliness, Target being the most steely and Chariot most mealy with Blenheim being intermediate. These structural differences were found to be associated with differences in the concentration of endosperm components, particularly proteins and β‐glucan. Analysis of nitrogen within the endosperm showed that protein was mainly concentrated in the embryo and distal regions with the inner, mid‐endosperm containing lowest levels. As the total nitrogen (TN) of the grain increased, the mealier samples accumulated nitrogen mainly in the embryo whereas the steely sample had higher levels in the central endosperm. SDS‐PAGE showed no differences in the protein banding pattern at different TN levels. Electron microscopy using immuno‐gold labelling demonstrated that γ‐hordeins were present in sub‐aleurone and outer endosperm whereas the C‐hordeins were found throughout the central endosperm. However, steely areas of central endosperm contained γ‐hordeins. During malting, protein modification in Chariot was more extensive than in Target with 34kD and 97kD hordeins being completely degraded. In Chariot and Blenheim, level of β‐glucan was low and it was evenly distributed throughout the endosperm. In the steelier Target, however, the amount of β‐glucan was higher and was concentrated in the proximal and distal areas of the endosperm. Steely grains (containing high concentrations of protein and β‐glucan) displayed slower water distribution during steeping and later development and distribution of β‐glucanase during germination. As a consequence, the steely sample achieved a lower degree of modification during malting. The structure of the endosperm, therefore, has a prime influence on the evenness of distribution of moisture and enzymes which is crucial for homogeneous modification during malting. © 1999 Society of Chemical Industry     

Effects of Mashing Parameters on Mash β‐Glucan,FAN and Soluble Extract Levels

To gain further technological knowledge of mashing, pilot scale mashing trials were carried out varying mashing programme (upward/isothermal mashing), milling procedure, grist:liquor ratio, time of mash stands, and grist modification level (well and poorly modified malt). During mashing β‐glucan, free amino nitrogen (FAN) and extract contents were analysed as key indicators for cytolysis, proteolysis, and amylolysis, respectively. The malt modification was of major impact for the β‐glucan release in contrast to a variation of milling procedure and of grist:liquor ratio. Extended stands lead to increased final values only for poorly modified malt. Similarly, FAN release was predetermined by malt modification while variation of milling and of grist:liquor ratio was not relevant in contrast to stand extension. None of the variations applied influenced extract yield as long as gelatinization temperature was reached. Greatest gains occurred around 57°C. In conclusion, wort quality is critically determined by malt modification. Mashing with well modified malt in combination with short stands should result in a mash of low β‐glucan and sufficient FAN level without losing extract yield. However, for poorly modified malt the variation of mashing parameters has an impact on the key indicators in which cytolysis plays the dominating role.     

Analysis of β‐Glucan in Single Grains of Barley and Malt Using NIR‐Spectroscopy

This paper describes initial experiments carried out in a collaborative study with Perten Instruments, Sweden, using Near‐Infrared spectroscopy to assess β‐glucan content in single grains of barley and malt. In general, the method needs further development, but this study shows that it has potential as a valuable tool for assessing endosperm modification of malt. The method is fast and non‐destructive and therefore allows other parameters related to endosperm modification to be analysed using the same grains.     

A Reappraisal of the Differences Between Scandinavian and Spanish Barleys: Effect of β‐Glucan Content and Degradation on Malt Extract Yield in the cv. Scarlett

Forty one samples of the malting barley cultivar Scarlett were collected from both Scandinavia (15 from Finland and 10 from Denmark) and the Iberian Peninsula (15 from Spain and 1 from Portugal), during the harvest years of 1998 and 1999. These samples were subjected to grain analyses, comprising protein content, hordein fractions by high performance liquid chromatography (HPLC) and β‐glucan content. The samples were micro‐malted and the malts were analysed to determine different patterns in the influence of grain composition on malt extract development linked to the two contrasting environments. The most obvious difference found between the Scandinavian and Iberian barleys was the effect of the total and insoluble barley β‐glucans. They were an effective barrier of malt extract in the North, but appeared to increase extract in the South. A conclusion was that the positive effect of β‐glucans in the Iberian barleys was a consequence of their greater capacity to synthesise and release β‐glucan hydrolases during germination.     

Malting Behaviour of Barleys Grown in Canada and Spain as Related to Hordein and Enzyme Content

To continue our effort to analyse the genetic (varietal) and environmental (sites and years) effects on malting quality of barley, we have field‐tested four barley varieties, two‐ and six‐rowed, European and North American, in Spain and Canada in 1998 and 1999. The Spanish trials were autumn‐sown whereas the Canadian ones were spring‐sown. Barley grain was analysed for total protein and hordein contents and micromalted. Canadian‐grown barleys had significantly lower contents of grain protein and all‐three hordein fractions than the Spanish ones. They also had lower malt respiratory losses, wort β‐glucan and viscosity but lower fine‐ and coarse‐ground malt extract yield, friability, free amino nitrogen, Kolbach index, α‐amylase and diastatic power. In other words, the Canadian‐grown barleys, despite showing lower protein and hordein contents, produced malt of inferior quality than their Spanish counterparts, which, overall, produced higher quantities of degrading enzymes (amylolytic, proteolytic and cytolytic) during germination, thus being able to attain higher extract yield levels.     

Optimization of Malting Conditions for Two Black Rice Varieties,Black Non‐Waxy Rice and Black Waxy Rice (Oryza sativa L. Indica)

Two black rice varieties, “black non‐waxy” and “black waxy”, were investigated as possible raw materials for the production of malt. The malting conditions were optimised using response surface methodology. The three process parameters were steeping, germination time and temperature. Each parameter was tested at three levels: adjustment degrees of steeping were 38, 41, and 44%, germination times were 6, 7, and 8 days, and the temperatures were 20, 25 and 30°C. At the end of the germination process, all samples were kilned at 50°C for 24 h, and shoot/rootlets were removed before a detailed quality assessment was performed. Data analysis was performed using the Design Expert Statistic Program. The optimal conditions found for both rice varieties were as follows: germination time of 8 days at 30°C and 44% grain moisture. Although the extract yield, and a‐amylase and β‐amylase activities of both rice malts were lower than barley malt, the higher activity of limit‐dextrinase enzyme and apparent attenuation limit (AAL), which was higher than 80%, suggests that rice malt has potential for use in brewing.     

Variety and germination time effect on total β‐glucan,water‐insoluble β‐glucan,water‐soluble β‐glucan components and β‐glucanase levels in improved sorghum varieties SK5912, KSV8 and ICSV400 before and after malting and their relationships to wort viscosity

The effects of variety and germination time on β‐glucan components – total β‐glucan (TBG), water insoluble β‐glucan (WIBG) and water soluble β‐glucan (WSBG) and β‐glucanase (BG) levels – before and after malting in improved sorghum varieties SK5912, KSV8 and ICSV400 and their relationships to wort specific viscosity (SV) were studied. This study was part of efforts to aid local malting and brewing industries in the application of sorghum varieties that are abundantly available to reduce costs. At the fifth day of germination, variety ICSV400 had the lowest TBG, WIBG and WSBG levels in its raw and malt samples. Variety SK5912 had the highest TBG, WIBG and WSBG levels in its raw samples, while variety KSV8 had the highest levels of TBG, WIBG and WSBG in its malt samples. Similarly, variety ICSV400 malts developed the highest BG levels, while the KSV8 malts gave the lowest level. The effect of variety, germination time and variety × germination time interaction was significant (p < 0.05) on the TBG, WIBG and BG levels and was not significant on the WSBG levels. Weak and significant correlation of TBG levels with SV (0.25, p < 0.05 for SK5912; 0.24, p < 0.05 for KSV8; and 0.31, p < 0.05 for ICSV400) was observed in all the samples, suggesting that the low β‐glucan levels may not be primarily and solely responsible for any viscosity impediments associated with sorghum worts during run‐off. With improvement in the effective utilization of sorghum, ICSV400 appeared the most suitable variety for malting and brewing in Nigeria.Copyright © 2016 The Institute of Brewing & Distilling     

Importance of Endosperm Modification for Malt Wort Fermentability1

Malt wort fermentability is dependent on an adequate supply of the essential nutrients required by yeast. The barley and malt factors affecting this supply of nutrients are not well understood. This study used two doubled haploid populations (Arapiles × Franklin, TR251 × HB345), the latter with a hulless barley parent, to investigate effects of barley and malt quality on fermentability. Populations were grown and malted at different locations resulting in a broad range in malt quality with significant differences in extract, modification and enzyme levels. Fermentability, as indicated by apparent attenuation limit and ethanol levels, also showed significant differences among samples. Modification was the most important factor for good fermentability. There appeared to be several different mechanisms by which modification affected fermentability. High viscosities, slow starch release during mashing, increased glucose supply from better β‐glucan breakdown and increased free amino nitrogen levels all affected fermentability. Effects of starch‐degrading enzymes on fermentability became more significant in better modified malts with α‐amylase showing stronger effects than diastatic power. The poorer fermentability of hulless barley malt was predominately due to low levels of α‐amylase, although, free amino nitrogen also appeared to be an important factor.     

Effects of seeding rate, nitrogen rate and cultivar on barley malt quality

BACKGROUND: Crop management tools have been shown to affect barley kernel size and grain protein content, but the direct effect on malt quality is not well understood. The present study investigated the effect of seeding rate, nitrogen fertilisation and cultivar on malt quality. RESULTS: Higher seeding rates produced barley with less grain protein and smaller, more uniformly sized kernels. The small, uniformly sized kernels modified more completely, leading to malt with higher extract and lower wort β‐glucan than malt from low‐seeding‐rate barley. Increasing rates of nitrogen fertilisation caused grain protein levels to increase, which limited endosperm modification and reduced malt extract levels. AC Metcalfe showed better modification and higher malt extract than CDC Copeland, but CDC Copeland had better protein modification at higher fertilisation rates, which resulted in less reduction of malt extract as nitrogen rate increased. CONCLUSION: Higher seeding rates reduced kernel size and grain protein levels without compromising malt extract owing to better endosperm modification of the more uniformly sized kernels. Negative effects of higher nitrogen rates on malt quality can be reduced through development of cultivars with improved ability to modify protein during malting. © 2012 Her Majesty the Queen in Right of Canada     

Effect of Different Steeping Conditions on Endosperm Modification and Quality of Distilling Malt

This study showed that when barley was steeped in water for either 8 h or 16 h, hydration of endosperm materials was suboptimal and modification of endosperm materials of barley malt was inadequate. The malt produced under these steeping regimes gave poor friability scores and produced a large number of whole grains. When barley was steeped for 24 h on a continuous basis, or when a regimented standard steeping method was used, the malt produced gave higher friability scores and a much lower number of whole grains. An important relationship was found between friability scores and whole grain results for the malt samples produced under these conditions. Optic barley, whose endosperm was more difficult to hydrate, gave a strong negative correlation between friability scores and number of whole grains at R² = 0.8689. Oxbridge, whose endosperm was more easily hydrated, gave a much stronger negative correlation between friability scores and number of whole grains at R² = 0.9769. Rapid visco‐analysis (RVA) results also confirmed that steeping the barley samples for only 8 h or 16 h produced malt that modified poorly as the RVA peak viscosities were very high. RVA pasting results further confirmed that when barley was steeped for 24 h on a continuous basis, or when a standard regimented steeping method was used, good quality malt was produced and no differences were found in the RVA peak viscosities of the barley malt samples produced under the two different steeping conditions. The results of protein breakdown (proteolysis) during these experiments, measured in terms of total soluble nitrogen (TSN) production, or soluble nitrogen ratio (SNR) further confirmed that optimal proteolysis was achieved when barley was steeped for either 24 h on a continuous basis, or with a standard steep. Optimal results were also found for hot water ex‐tractable materials such as hot water extract (HWE) and free amino nitrogen (FAN) when barley was steeped for 24 h or the standard steep. The 24 h continuous steep for barley produced quality malt comparable to that obtained when the standard regimented steep was used for steeping barley. For both Optic and Oxbridge barley, with a 24 h continuous steep, produced malt that gave significantly higher fermentabilities and PSY values, regardless of germination time, than those obtained using a standard regimented steep. Therefore steeping barley for 24 h on a continuous basis prior to malting will produce good quality malt for some barley samples/varieties. This will help to reduce water usage during steeping, will save steeping time thereby reducing malting time and will reduce the amount of water for effluent treatment. All of these factors result in an overall cost saving for the malting industry.     

Refining the Prediction of Potential Malt Fermentability by Including an Assessment of Limit Dextrinase Thermostability and Additional Measures of Malt Modification,Using Two Different Methods for Multivariate Model Development

Prediction of malt fermentability (apparent attenuation limit — AAL) by measurement of the diastatic power enzymes (DPE), α‐amylase, total limit dextrinase, total β‐amylase, β‐amylase thermostability, and the Kolbach index (KI or free amino nitrogen — FAN) is superior to the conventional use of diastatic power (DP) alone. The thermostability of β‐amylase is known to be an important factor in determining fermentability, thus the thermostability of the other relatively thermolabile enzyme, limit dextrinase, was investigated to determine if it was also useful in predicting fermentability. To facilitate this aim, methods were developed for a rapid and cost efficient assay of both β‐amylase and limit dextrinase thermostability. Internationally important Australian and international malting varieties were compared for their total limit dextrinase and β‐amylase activity and thermostability. Interestingly, the level of limit dextrinase thermostability was observed to be inversely correlated with total limit dextrinase activity. The prediction of malt fermentability was achieved by both forward step‐wise multi‐linear regression (MLR) and the partial least squares (PLS) multivariate model development methods. Both methods produced similar identifications of the parameters predicting wort fermentability at similar levels of predictive power. Both models were substantially better at predicting fermentability than the traditional use of DP on its own. The emphasis of this study was on the identification of predictive factors that can be consistently used in models to predict fermentability, because the model parameter estimates will subtly vary depending on mashing conditions, yeast strain/fermentation conditions and malt source. The application of these multivariate model development methods (PLS and MLR) enabled the identification of further potential fermentability predicting factors. The analyses divided the predictive parameters into those defined by DP enzymes and those associated with modification (KI, FAN, fine/coarse difference, wort β‐glucan and friability). Surprisingly, limit dextrinase thermostability was not a substantial predictor of fermentability, presumably due to its negative correlation with total limit dextrinase activity. The application of these insights in the malting and brewing industries is expected to result in substantial improvements in brewing consistency and enable more specific quality targets for barley breeder's progeny selection cut‐off limits to be more precisely defined.     

Control of Superoxide Dismutase Activity during Malting Using Plackett‐Burman and Box‐Behnken Experimental Design and Its Effect on Reducing Power of Wort

The Plackett‐Burman multifactorial design was employed to screen the important malting parameters for superoxide dismutase (SOD) in final malt of Ganpi‐3. The eight factors screened for SOD were steeping temperature, steeping time, peroxide hydrogen concentration in steeping water, germination temperature, germination time, withering temperature, drying temperature and kilning temperature. Variance analysis showed that steeping time, germination temperature and kilning temperature were significant for SOD activity. Box‐Behnken experimental design was further used to optimize the levels of the above three factors. By response surface methodology and canonical analysis, the optimal malting factors for higher SOD activity in final malt were: steeping time 42.2 h, germination temperature 16.9°C and kilning temperature 82.9°C. Under these conditions, the model predicted a SOD activity of 2234 U/g of dry weight malt. Verification of the optimization showed that a SOD activity of 2220 U/g was observed under optimal conditions. It showed that the experimental data could be reliably predicted by the polynomial model. Besides Ganpi‐3, three other barley varieties including Ganpi‐4, Ken‐2 and Hamelin were malted under optimal and common conditions under laboratory conditions. To some extent, SOD activities were higher in malts from the optimal malting process than those from the common malting process. Especially, SOD activities in Ganpi‐3 and Hamelin increased by 18.8% and 15.3%, respectively. Furthermore, twenty‐nine samples of malts, including eleven imported malts and eighteen domestic malts, were used. Relationships between SOD activity in malt and the reducing power of wort were examined. There was significant correlation between SOD activity and the reducing power of wort (R² = 0.8069).     

The effect of germination time on malt quality of six sorghum (Sorghum bicolor) varieties grown at Melkassa,Ethiopia

The effect of germination time (48, 96 and 144 h) on malt quality of six sorghum varieties was investigated to determine the potential of grain sorghum cultivars in the local brewery industry. Six sorghum varieties (Gambella 1107, Macia, Meko, Red‐Swazi, Teshale and 76 T1#23) were evaluated in a randomized complete block design with three replications. The results showed that both germination time and variety had a significant effect (p ≤ 0.05) on sorghum malt quality. The hectolitre weight (kg hL^?1 at 12.5% moisture), germination energy (%), crude protein (%) and flour starch amylose content (%) were: 75.8–82.9, 96–99, 7.0 ? 11.9 and 16.0 ? 23.0, respectively. The sorghum malt diastatic power (DP, °L), free amino nitrogen (FAN, mg L^?1), hot water extract (HWE, %) and malting loss (%) were: 18.96 ? 31.39, 185.67 ? 343.29, 41.85 ? 85.08 and 8.68 ? 27.56, respectively. Malting loss, HWE and FAN increased with germination time. The DP increased as the germination duration increased from 48 to 96 h, but the difference between 96 and 144 h was not significant. Considering the excessive malting loss and marginal increase in HWE beyond 96 h, this study suggests that the optimum malting duration would be around 96 h. Among the varieties tested, Teshale and Gambella 1107 produced the better malt quality. Copyright © 2012 The Institute of Brewing & Distilling     

CHANGES IN ACTIVITY OF SORGHUM LIPASE DURING MALTING AND MASHING

Lipase activity was monitored during malting and mashing of sorghum grains. All three sorghum varieties contained detectable lipase activity in the ungerminated form. Lipase activity changed only slightly during steeping for 24 hours but increased several fold in the course of germination. Between 24% and 60% of the lipase activity of the green malt was retained after kilning at 48°C but no activity was detected in the wort after mashing at 65°C. About 68% of the lipase activity of 72 hours old malt was detected in the plumule, while 29% and 3% were detected in the endosperm and radicle, respectively. Optimum activity was observed at pH 7.0.     

Effect of Malting Conditions on Pearl Millet Malt Quality

The effect of malting conditions on pearl millet malt quality in two varieties, SDMV 89004 and SDMV 91018, was investigated. Grain was steeped and germinated at four temperatures, 20°, 25°, 30° and 35°C, over 5 days. Generally, malt quality parameters (percentage of roots and shoots, diastatic power (DP), α‐ and β‐amylase activity, free α‐amino nitrogen (FAN), and malting loss) were significantly affected (P < 0.001) by germination temperature and time, as well as by variety. Malt FAN and malting loss were not affected by variety. A germination temperature of 25–30°C and germination time of 3–5 days were optimal. These conditions resulted in high DP, α‐ and β‐amylase activity, good FAN and moderate malting loss. These malting conditions and the subsequent malt quality of pearl millet are similar to those reported for sorghum. Pearl millet malt can therefore be used for the production of sorghum type beers.