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.     

Multiple Linear Regression Calibrations for Barley and Malt Protein Based on the Spectra of Hordein

The feasibility of using NIR spectral information from barley and malt hordein was assessed as to the suitability of developing improved NIR calibrations to predict protein in barley and malt. Using extracted hordein it was possible to gain more information on wavelengths relevant to predict protein with reduced errors. Strong correlations for grain protein and NIR wavelengths were found at 1,116, 1,268, 2,040, 2,068, 2,188 and 2,300 nm. Multiple linear regression equations provided improved predicting power for barley and malt protein with a standard error of prediction of 0.15 and 0.17%, respectively, whereas partial least squares regression gave a standard error of prediction of 0.22 and 0.27% for barley and malt, respectively. The use of NIR becomes more pronounced in breeding programs as NIR is a rapid and non‐destructive technique allowing the screening of early generation lines with limited grain quantities. Also, the spectral analysis of native components from resting grain components will assist in building calibrations that provide qualitative values rather than just ranking breeding lines.     

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.     

HIGH PERFORMANCE LIQUID CHROMATOGRAPHY OF BARLEY PROTEINS: RELATIVE QUANTITIES OF HORDEIN FRACTIONS CORRELATE WITH MALT EXTRACT

Several high performance liquid chromatography (HPLC) approaches have been used to study the relationships between amounts of particular groups of grain storage proteins (hordeins) and malt extract, using two sets of barley samples. The areas of several chromatogram peaks varied between samples in a manner that correlated with malt extract. Size-exclusion HPLC of protein aggregates extracted by sonication gave negative correlations of the relative areas of the three largest chromatogram peaks with malt extract in a set of 39 samples of the one variety, but not in a set of samples of 8 varieties grown at 4–5 sites. Under reducing conditions, disulphide-bonded hordeins were quantitatively extracted and the proportion of hordein in the largest peak, comprised of D- and B-hordeins, correlated well with malt extract in both sets of samples. The hordeins sequentially extracted into 50% (v/v) 1-propanol and 50% (v/v) 1-propanol-50 mM dithiothreitol (DTT) were also analysed by reversed-phase HPLC, and could be resolved into several peaks of B-, C- and D- hordeins. Relationships between the amount of certain C- hordeins in the propanol extract, and certain B- hordeins in the propanol-DTT extract and malt extract were seen. As well as providing a better understanding of the relationship between barley grain protein composition and malting quality, these HPLC methods are useful additional methods to enable partial prediction of malt extract using un-malted barley.     

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.     

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.     

MEASUREMENT OF DIASTATIC POWER

Modifications to the method of estimating the diastatic power of malt using starch hydrolysis and determination of reducing sugar released with p-hydroxybenzoic acid hydrazide are described. Using 40 g malt, 4% w/v soluble starch reduced with sodium borohydride and a “boiled enzyme blank”, the production of reducing sugar is directly proportional to volume of added extract. Using a constant amount of extract, the reaction is directly proportional to time over a period of 30 min. The method enables the direct estimation of the diastatic power of malts with Windisch-Kohlbach values of up to 434 degrees.     

Effects of Wheat Protein Content on Endosperm Composites and Malt Quality

In this study, the malting performance of six traditional Chinese wheat varieties was evaluated. The effects of protein content on wheat endosperm composition and the malt quality were studied. The β‐glucan in the malt showed a positive correlation (r = 0.806, P<0.1) with the protein content of the wheat. With an increase in protein content, there was a significant linear decrease in extract (r = ?0.923, P<0.01) and Kolbach Index (r = ?0.850, P<0.05), but diastatic power, soluble protein and FAN gave a peak value at 16.0% protein content. Because of the suitable protein and starch content, the malt qualities of wheat W‐3 were satisfactory. Extract was 81.7%, saccharification time was 6.5 min, diastatic power was 484.07WK, FAN was 136.58 mg/100 g, the Kolbach index was 40.0% and viscosity was 1.77 cP.     

FACTORS GOVERNING THE MILLING OF MALT

A method has been developed using a laboratory 2 roll mill for reproducibly milling malt produced from European Barley (two-row). The procedure can be readily adapted to give grists of similar particle size distribution to those produced by either commercial four or six roll mills. Factors which were considered likely to affect the milling of malt were studied with particular attention to moisture level, the degree of modification, extent of kilning, influence of storage, differences in grain size and extent of steam conditioning. Most of these factors did exert an effect on milling. However, in the majority of cases the values needed to be outside normal commercial limits before substantial changes in milling performance were observed. Studies employing a laboratory mashing system have shown that by using malts containing c.a. 6% moisture it is possible to reduce wort separation times without affecting extract recovery. Steam conditioning can have a marked beneficial effect on the milling performance of malt providing the extent of moisture pick-up is carefully controlled.     

Simultaneous estimation of amylose,resistant, and digestible starch in pea flour by visible and near-infrared reflectance spectroscopy

Due to its health benefits, resistant starch (RS) has received increasing attention from the public, and there is a need to develop methods to measure the amylose and RS concentration in pea (Pisum sativum L.) flour. The aim of this study was to develop a visible and near-infrared reflectance (vis–NIR) model for the simultaneous determination of amylose, RS, and digestible starch (DS) in pea flour. A total of 123 dry pea samples consisting of different pea varieties grown in different environments were collected, and ground to flour, and then the vis–NIR spectra were scanned. The amylose, RS, and DS contents of the pea flours were also measured by an enzymatic colorimetric assay. The spectra data were calibrated with the enzymatic colorimetric-assayed values. Results showed that amylose, RS, and DS in the pea flours can be simultaneously estimated using the vis–NIR spectra. Instead of using the full spectrum (300–2300 nm), we found the most efficient wave bands lying in the visible region between 370 and 560 nm and the NIR spectra in the range of 1600–1800 nm. Using the stepwise regression with backward elimination method, the multiple linear regression (MLR) models were developed from the most efficient wavelengths. The MLR models had the determination coefficients R² of 0.95, 0.76, 0.80, and 0.88 for amylose, RS, DS, and total starch, respectively. The correlation coefficients between model estimated and the enzymatic colorimetric assayed values were 0.97, 0.80, 0.85, and 0.93 for amylose, RS, DS, and total starch, respectively.     

Near Infra-red Monitoring of Enzymatic Hydrolysis of Starch

NIR measurement was carried out for a starch dispersion circulating from a reactor to an NIR flow-through cell over a wavelength range of from 1000 to 2500 nm in 2 nm increments. Multiple regression analysis between NIR and HPLC shows a good correlation. It is shown that the NIR method can be used to determine glucose quantitatively in real time during enzymatic hydrolysis, and that NIR may be employed for controlling the reaction in a bioreactor.     

EVALUATION OF THE INFRAALYZER 400 FOR THE ANALYSIS OF BARLEY AND MALT

A collaborative study is described of the use of infra-red reflectance for the evaluation of the analytical characteristics of barley and malt. Transfer of centrally-prepared calibrations to other reflectance machines was found to be possible for moisture content and protein content in barley and malt but not for malt extract or malt modification. The examination of infra-red reflectance results, obtained at different wavelengths, by step-wise ascending regression rather than step-wise descending regression showed that the former was more satisfactory, especially in being more rapid in execution.     

Application of Biological Acidification to Improve the Quality and Processability of Wort Produced from 50% Raw Barley

In this study four strains of lactic acid bacteria (LAB) were chosen to bioacidify a mash containing 50% barley and 50% malt. The strains were isolated from malted and unmalted barley and assayed for extracellular enzymatic activities (proteases, amylases, β‐glucanases). The biologically acidified mash was compared to a chemically acidified mash, 100% malt mash un‐acidified and 50% malt and 50% barley mash unacidified. Characteristics such as pH, extract, colour, viscosity, total soluble nitrogen (TSN), free amino nitrogen (FAN), apparent fermentability, β‐glucan and lautering performance of the resultant worts were determined. A model lautering system replicating one used in a brewery was designed and built in University College Cork (UCC) to measure the lautering performance of the bioacidified mashes. The new system was compared to the filtration method used in EBC method 4.5.1. Overall the addition of LAB to bioacidify a mash of 50% barley and 50% malt resulted in faster filtration times, which correlated with decreased β‐glucan levels. Proteolytic LAB had a positive influence on the quality of wort and resulted in increased FAN levels. Lighter colour worts were observed along with increased extract levels.     

GRAIN AND MALT MILLING ENERGIES IN SORGHUM AND THEIR RELATIONSHIPS WITH EXTRACT AND DIASTATIC POWER

Twenty three sorghum genotypes were shown to vary widely for a range of grain and malt quality characters. Grain milling energy did not give an indication of the likely malting performance of sorghum samples. There were, however, close relationships between malt milling energy and both % extract and diastatic power, if samples with unusually high or low grain nitrogen contents were excluded from the population under study. Rapid screening of very small malt samples for milling energy and diastatic power could form the basis for malting quality selection in early generations of sorghum breeding programmes.     

THE IN VIVO AND IN VITRO DEGRADATION OF BARLEY AND MALT STARCH GRANULES

The susceptibilities to amylolytic hydrolysis of the two different types of starch granule in barley and malt have been investigated. The large and small granules from both germinated and ungerminated grain were subjected to the sole action of malt α-amylase under conditions which otherwise simulated those of a conventional infusion mash. Large starch granules from barley are hydrolysed at a slower rate than those from malt. The faster conversion of the latter is attributed to prior modification of the starch granule structure during germination. The small granules from barley are extremely resistant to attack by α-amylase and even pre-cooking does not appreciably increase their susceptibility to amylase attack. Although the small starch granules from malt are less susceptible than the corresponding large granule fraction, they can be hydrolysed to a greater extent than can the small granules from barley. The increased susceptibility of small granules from malt is probably due to the partial removal of their protecting surface layer of protein. Although during malting the small granules of barley are hydrolysed more rapidly than the large granules, the situation is reversed during mashing. Very little loss of extract can be attributed to the enzymic resistance of small starch granules in all-malt mashes. If raw barley is used in the grist, substantial amounts of small starch granules may remain in the mash.     

Application of high‐pressure treatment in the mashing of white malt in the elaboration process of beer

In this study, high‐pressure treatment (HPT) was applied to the mashing stage of beer production, which involves drying and milling of white malt and subsequent mixing with water. The following parameters were evaluated after pressurisation: β‐glucanase activity, starch gelatinisation and sugar extraction. Evaluation of starch hydrolysis from the malted barley endosperm after HPT was performed by measuring β‐glucanase activity after pressurisation; this enzyme breaks down gums and β‐glucans in wort and is desirable to obtain a good‐quality beer. Soaked malt samples pressurised at 200–600 MPa showed no increase in this activity compared with controls. Conversion of milled malt was evaluated indirectly by measuring the gelatinisation of starch, which began at 400 MPa. Soluble sugars were also measured in pressurised samples from the mashed liquid to investigate saccharification during the mashing stage. After 400 or 600 MPa treatment for 20 min, both the sucrose (g per 100 ml) and extract (l ° kg^?1) values were the same as those found in mashed samples following the standard procedure used in the brewing industry (65 °C,90 min). Starch gelatinisation was analysed at different high pressures (200–600 MPa) and it was shown that gelatinisation began at 400 MPa. The HPT time would have to be shorter to make the process commercially attractive. © 2002 Society of Chemical Industry     

USE OF THE HAGBERG FALLING NUMBER APPARATUS TO DETERMINE MALT AND BARLEY QUALITY

The Hagberg Falling Number measurement, widely used in the milling industry for assessing the baking quality of wheat flour, has been modified and used as a rapid mashing technique to assess malt quality. Barley Malt flour is mixed with cold mashing liquor containing 0.1% calcium chloride and 15 mM beta-mercapto-ethanol. The slurry is heated and mixed by the Falling Number apparatus, and the starch is gelatinized. After one minute of heating/mixing the stirring rods are raised and released. As the gelatinized starch is hydrolysed by amylases, the viscosity of the slurry decreases allowing the stirring rods to fall. The time taken for these rods to fall a fixed distance is called the Falling Number. A relationship has been determined between the Falling Number obtained for a malt and the quality of the malt. Poor quality malts had a high Falling Number (e.g. >200s) whilst good malts had a low Falling Number (e.g. <150s). The correlation coefficient between Falling Number and Hot Water Extract (IOB coarse grind) was ?0.89, and between Falling Number and Friability was ?0.96. These good correlations indicate the measurement was influenced by the degree of endosperm modification. Being a mashing technique, however, the Falling Number was also influenced by other factors which influence mashing such as enzyme complement and water binding. This technique has the potential to measure malt quality using a commercial apparatus more rapidly than present malt analyses. No significant relationship could be determined between the Falling Number and the malting quality of barley.     

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.