Re‐Assessment of the Half‐Grain Modification Method for Assessing Malt Modification

The half‐grain mashing (modification) method proposed by Palmer (J. Inst. Brew., 1975, 81: 408) was reassessed. The intention was to quantify the differences in malt modification in terms of β‐glucan breakdown and clarify the relationship between β‐glucan breakdown and overall modification of the endosperm during malting. This was carried out at 45°C as well as at 65°C, the percentage of weight loss was recorded and the endosperm residue was analysed for β‐glucan content. In general, weight loss was related to modification. Samples, which were modified at higher levels, lost significantly more material during the half‐grain mashing procedure than those which were under‐modified. At a malting process time of 96 h all the varieties had similar weight loss. After mashing the half grains, the β‐glucan contents of the grain residues showed an apparent increase because of loss of non‐β‐glucan materials. However, over the malting period β‐glucan decreased. Chariot malted faster than the other varieties studied. The β‐glucan levels of this variety were reduced by 78% between 48 and 72 h of germination. Significant levels of β‐glucan were degraded and large quantities of starch and protein were released. During the same period of germination, the corresponding samples of Decanter did not show a significant reduction in β‐glucan levels. In contrast, Brazilian variety MN698 lost endosperm material and β‐glucan rapidly by 48 h. These early results suggest that during malting, extract solubilization may or may not accompany β‐glucan breakdown. Therefore, β‐glucan levels in malt cannot be used as an overall index of modification of the endosperm.     

Characterization of Solubilized Forms of Bound ß‐Amylase Released by Various Agents

In a previous study the patterns of release of bound β‐amylase caused by various agents were investigated. In the present study the released β‐amylase isoforms were purified and characterized. The agents used to release bound β‐amylase at different temperatures were papain, thiols, starchy endosperm extract and several substances with amphipathic characteristics. The isoforms released were compared to those present in soluble β‐amylase from 8‐day malt. They were characterized by sodium dodecyl sulphate‐polyacrylamide gel electrophoresis (SDS‐PAGE), isoelectric focusing (IEF), native polyacrylamide gel electrophoresis (PAGE), western blotting and attempts were made to use matrix assisted lazer desorption ionization (MALDI) mass spectrometry. β‐Amylase released by papain was similar in molecular weight (57 kDa by SDS‐PAGE) to soluble β‐amylase from 8‐day malt (56 kDa). The isoforms released by thiol reagents were larger (57–58 and 60–62 kDa) and were similar to the soluble β‐amylase extracted from barley (56–57 and 60–61 kDa). Agents with amphipathic characteristics (bovine serum albumin and the detergents CHAPS and Triton X‐100) released higher molecular weight material (62 kDa). Extracts from starchy endosperm containing proteolytic activity released a β‐amylase isoform, intermediate in size (58 kDa) between that released by papain and those released by thiol‐containing and amphipathic agents. Heat‐treated starchy endosperm extract (lacking protease activity) caused limited (15%) release of β‐amylase isoforms (58 and 61 kDa). These resembled the isoforms released by thiol and hydrophobic agents. The β‐amylase isoforms exhibited heterogeneity when analyzed by IEF, native PAGE and MALDI. The results confirm that a complex combination of mechanisms is involved in the release of bound β‐amylase during the germination of barley.     

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     

Changes in β-glucan and other carbohydrate components of barley during malting

Changes in total (1→3), (1→4)-β-glucan content were followed during the micro-malting of nine varieties of barley with a wide range of malting qualities. These changes were related to estimates of endosperm modification based upon staining with Calcofluor. β-Glucan content declined from an average of 3.54% in the barley to 0.75% in the malt. Pentosan and total starch (including starch-derived oligosaccharides) levels showed comparatively little change during malting. β-Glucan composition of the barley was a poor indicator of malting performance. However, the β-glucan, starch and xylose contents of the malt all showed significant correlations with malt extract. Estimation of malt β-glucan content gave the best indication of malt quality. Direct determination of β-glucan may be of more value in assessing malt quality than indirect techniques based upon assessing modification of stained grains.     

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.     

EVALUATION OF BARLEY AND MALT QUALITY USING NEAR-INFRARED REFLECTANCE TECHNIQUES

A scanning near-infrared reflectance spectrophotometer was calibrated for the prediction of barley aleurone colour and malt moisture. The malt moisture was predicted on malt ground for the determination of malt extract (coarse grind) making the method suitable for moisture correction in malt extract estimation. Calibrations for the prediction of malt extract and endosperm modification from barley and malt were also attempted. A correlation (r= 0.851 n = 135) was found between malt hot water extract and the percentage of the endosperm estimated as being modified by microscopy following staining with Calcofluor. Probably because of this influence of modification on malt extract, the use of near-infrared reflectance to predict malt extract was most successful at predicting the malt extract values obtained following micro-malting in the absence of the additives, gibberellic acid and potassium bromate.     

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.     

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.     

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.     

Enhancing the levels of 4‐vinylguaiacol and 4‐vinylphenol in pilot‐scale top‐fermented wheat beers by response surface methodology

Top‐fermented wheat beers are desired by consumers because of their distinctive clove‐like and phenolic aroma, in which two characteristic flavour substances, 4‐vinylguaiacol (4VG) and 4‐vinylphenol (4VP), are of vital importance. In this paper, the levels of 4VG and 4VP were the subject of an intensive and detailed investigation. The influence of wheat malt proportion, mashing‐in temperature, boiling time and fermentation temperature on 4VG and 4VP was studied by pilot‐scale brewing experiments using a 10 hL mashing vessel and 20 hL fermentation tanks. Response surface methodology (RSM) was used to explore the possibilities for enhancing the concentration of 4VG and 4VP in top‐fermented wheat beers. Statistical analysis of the results showed that for the ranges of wheat malt proportion studied, mashing‐in temperature, boiling time and fermentation temperature all had a significant influence (p <0.05) on 4VG and 4VP production. Based on the response surface plots, the optimal brewing parameters that maximized 4VG and 4VP levels were as follows: (a) wheat malt proportion 40%; (b) mashing‐in temperature 44°C; (c) boiling time 88 min; and (d) fermentation temperature 19.5°C, resulting in significantly increased levels of 4VG and 4VP (2.418 mg/L and 1.402 mg/L, respectively). These brewing and fermentation parameters were concluded to be the optimal conditions to highlight the typical flavour and aroma of top‐fermented wheat beers. Copyright © 2015 The Institute of Brewing & Distilling     

Malting of Sorghum: Further Studies on Factors influencing α‐Amylase Activity

The effects of steep regime, nature of alkaline steeping agent, and kilning condition on α‐amylase development were studied for four Nigerian sorghum cultivars. Malt α‐amylase activity was highly significantly (p<.001) influenced by all the four factors as well as their various assortments of interaction. Generally malts from the Local Red (LR) variety produced the highest a‐amylase values, followed by those of SK 5912, Local White and KSV 8 in the above sequence. The presence/absence of air‐rest processes in steep regimes was a significant factor (p<.001) influencing malt α‐amylase response to final warm steeping as well as to the other factors under study. Similarly, the nature of the steeping agent was a very significant determinant of malt α‐amylase response to kilning condition and regime of steeping. Of significant interest was the observation that Ca (OH)₂ steeps enhanced malt α‐amylase activity at the higher temperature of kilning. The significantly lower α‐amylase values given under similar conditions by the other alkaline liquors suggest a possible increase in malt thermostability due to steeping in Ca (OH)₂. Additionally, the fact that the extent of enhancement of malt α‐amylase activity by Ca (OH)₂, at 50°C Kiln temperature, was regime‐dependent, suggests that the latter was an important modulator of sorghum germination physiology.     

Brewing with Rice Malt — A Gluten‐free Alternative

The preparation of beer‐like beverages with rice malt as the only raw material is reported. Several tests were performed on a laboratory scale and in a 25 L‐capacity pilot plant. Both the decoction and the infusion procedure were tested; malt and water were mixed in a ratio 1:3.5 for both methods and the mash was brewed without adding exogenous enzymes. The obtained worts were fermented using bottom fermenting yeasts, while “beers” were re‐fermented utilizing top fermenting yeasts and adding either sterile wort or sugar. A maximum ethanol of 4.5% vol. was obtained after the primary fermentation from an initial wort with an original gravity of 11.8°Plato. All parameters of the beer were found to be acceptable using a standard beer analysis. Owing to a suitable hop addition, an aroma very similar to that of a normal beer was obtained.     

HOMOGENEITY OF THE FRIABLE FLOUR OF MALTING BARLEY

A detailed comparison was made of the properties of the friable flours and non-friable residues of two samples of malted barley of different nitrogen contents. The friable flours were sieved and fractionated to give a range of particle sizes, and the intact malt, whole friable flour, non-friable residues and fractionated friable flours were subjected to a range of analyses. Endosperm fractionation studies showed that the pattern of enzymic degradation of proteins in the modified friable flour of low nitrogen malt was more uniform than the corresponding pattern of protein breakdown in the friable flour of high nitrogen malt. Examination of the non friable residues showed that cell wall breakdown in the high nitrogen malt was less extensive than the low nitrogen malt. It is proposed that the high protein levels in the endosperm caused starch/protein compacting which limited endosperm hydration and enzymic modification during malting. The friability scores of high nitrogen malts may given an over estimate of endosperm modification.     

Mechanisms of the Release of Bound β‐Amylase

The aim was to identify the mechanism(s) responsible for the release of bound β‐amylase in soluble forms in vitro and during the germination of barley. Preparations of bound β‐amylase, from barley, were treated with different agents including papain, thiols, starchy endosperm extract and reagents with amphipathic characteristics. The time courses of the release of the bound enzyme were compared using the different releasing agents. All the reagents tested caused some release of bound β‐amylase. Probably a very complex combination of mechanisms is involved in the release of bound β‐amylase as barley germinates. Initial release may be by cleavage of disulphide bonds which bind β‐amylase to insoluble protein. Material having amphipathic characteristics may break hydrophobic associations between β‐amylase and insoluble protein(s). The larger molecular weight isoforms are proteolysed to form the lower molecular weight materials present in malt. Limited proteolysis may also occur before the release of bound enzyme. Whether or not proteolysis is directly responsible for the release of bound enzyme remains uncertain.     

Effect of batch‐to‐batch variation on the quality of laboratory and commercially malted Oxbridge barley

The malting performance of two different batches of Oxbridge barley was studied. The study showed that a single 24 h continuous laboratory steep was suitable for steeping the two batches of Oxbridge to provide good quality malt. Although each batch represented the same variety, and was of similar malting quality, they behaved differently in terms of friability scores, filtration rate and predicted spirit yield (PSY) by day 4 of the germination period. Worts prepared from the two malts gave different patterns of filtration rates, showing that each batch of Oxbridge modified at a different rate and in a different way, although they achieved similar final PSY results at the end of the malting period. The two batches of Oxbridge both malted rapidly, but behaved differently in terms of modification pattern, as revealed by the Rapid Visco‐Analysis peak viscosities, as well visual germination results. The results of this study confirmed that, given the same malting conditions, batch 1 germinated more rapidly than batch 2, and supports the view that Oxbridge shows some inconsistency as a malting barley variety. The study further showed that the PSY result per se is not necessarily a good indicator of optimum modification of finished malted barley. This study showed that, while a particular barley variety, such as Oxbridge, may be associated with certain qualities and can produce good quality malt when a short steeping cycle is used, other factors are likely to cause batch‐to‐batch variations from the same barley variety. While the short steeping cycle was very successful in producing high‐quality laboratory malt from Oxbridge, and results in a significant reduction of the quantity of water required for steeping, reduces malting time and also saves costs, there is evidence that there may be some additional variability associated with Oxbridge malting barley. Copyright © 2012 The Institute of Brewing & Distilling     

The influence of LOX‐less barley malt on the flavour stability of wort and beer

The aim of this study was to investigate the influence of lipoxygenase‐less (LOX‐less) barley malt on the quality of wort and beer, with the main focus on beer flavour stability. In the current study, pilot‐scale (1000 L) brewing trials were conducted with a control barley malt AC Metcalfe and a LOX‐less barley malt, PolarStar. The results clearly indicated that the LOX‐less barley malt showed less nonenal potential than the control, although LOX activities in both barley malts were relatively low. The beer brewed from the LOX‐less barley malt contained much lower concentrations of trans‐2‐nonenal (T2N) and gamma‐nonalactone, especially after the (forced or natural) aging of the beer, compared with the beer brewed under the same conditions using the control malt. The sensory panel evaluation indicated similar results in the general flavour profile. The freshness scores of beer brewed from the LOX‐less malt were higher than those from the control malt, and this was more pronounced after forced aging. In addition, the beer brewed from LOX‐less malt had a much better foam stability, almost 30 s (NIBEM test). These results confirm that the use of the LOX‐less barley malt was beneficial to beer flavour stability and foam stability. Copyright © 2014 The Institute of Brewing & Distilling     

The Relevance of Different Enzymes for the Hydrolysis of β‐glucans in Malting and Mashing

β‐Glucosidase and to a lesser extent endo‐β1–4‐glucanase are present in significant quantity in raw barley. These enzymes, as well as endo‐β1–3, 1–4‐glucanase, endo‐β1–3‐glucanase and exo‐β1–3‐glucanase, increase in activity during steeping and germination. Exo‐β1–3‐glucanase stands apart through its very late development during germination and it may be a limiting enzyme in malts that have not received substantial modification. Three separate exo‐glucanases were located in malt and each of them displayed a preference for β1–3 linkages. As the principle product of endo‐β1–3, 1–4‐glucanase is oligosaccharides with a β1–4 linkage at the non‐reducing end (from which end exo enzymes approach the substrate), this is likely to be a second reason (alongside the late development of the enzyme) why such oligosaccharides survive in significant quantities into wort. A third contributing factor may be the sensitivity of the exo‐glucanases to ions such as potassium, sodium and magnesium.     

Behaviour of Malted Cereals and Pseudo‐Cereals for Gluten‐Free Beer Production

Beer, a beverage frequently requested by celiac patients, is toxic to them due the gluten content derived from traditional raw materials used in the brewing process. To understand a possible alternative use of cereals and pseudo‐cereals, micro‐malting experiments on sorghum, buckwheat, quinoa and amaranth were performed using different malting parameters to study their brewing behaviour. Steeping in dilute NaOH solution was one parameter employed to understand any possible positive effect on malt quality. Alkaline steeping has been applied to pseudo‐cereals for the first time and often showed an increase in total soluble nitrogen (TSN) and free amino nitrogen (FAN), especially with buckwheat, where two results showed the TSN content increased by about 500 and 1,000 mg/L. An interesting result was found with amaranth fermentability, which showed a fermentability of 56%, higher than what was previously reported. From these positive results for malt quality, it is possible to affirm that the cereals and pseudo‐cereals investigated in this study could be employed for gluten‐free beer production. Furthermore, the alkaline steeping seems to be a useful process variable for optimisation of malt quality. Future studies about its effect on the diastatic power could be conducted.