AN ASSESSMENT OF THE MALTING QUALITY OF A MUTANT OF THE BARLEY CV. TROUBADOUR GROWN UNDER SCOTTISH CONDITIONS

The barley mutant, TR49, selected in Spain for rapid germination, and its parent cultivar Troubadour, were grown in Scotland in 1991 and assessed for a range of grain and malt quality characters. Results suggested that both genotypes had a potential dormancy problem, although TR49 showed a slight but significant improvement over Troubadour. TR49 produced very large grain which did not completely modify during malting, but still showed an advantage over its parent cultivar for all malting parameters assessed.     

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.     

EVALUATION OF A MICRO-MALTING PROCEDURE USED TO AID A PLANT BREEDING PROGRAMME

A micro-malting and a malt analysis procedure, using less than 40 g of seed, is described and critically appraised as a means of assessing the malting potential of samples generated in a barley breeding programme. Four barley varieties representing a range of malting abilities were used to evaluate the method. The magnitude of variances due to analyst, batch and cumulative experimental errors were compared to variation attributable to variety for 22 characters examined during seed preparation, malt production and malt analysis. For most characters almost all of the variance was due to variety; these included hot water extract, soluble nitrogen, cold water extract, grain nitrogen and thousand corn weight, which are the characters principally used for assessing malting ability. The results show that the procedure is suitable for determining the grain characters and malting potential of the large numbers of samples submitted from breeding trials and also draw attention to those requiring particular care in defining analytical techniques and requiring consistency in application.     

EFFECTS OF AIR REST PERIODS ON MALTING SORGHUM RESPONSE TO FINAL WARM WATER STEEP

The effects off various lengths of air rests on response off malting sorghum main root length, malting loss, diastatic activity, α- and β-amyloytic activity and extract to final warm water steep at 40°C was investigated. Grains were steeped in distilled water for a total of 48 h at 30°C with a 6 h final warm water steep under four different cycles incorporating 1, 2, 3 or 4 h air rest periods. All malt quality properties evaluated were significantly affected by the length of air rest, the cultivar and their pair-wise interactions. Main rootlet length and malting loss reduced progressively as the length of air rest increased. Reducing power, α- and β-amylolytic activity and extract generally increased as the length of air rest was increased for both sorghum cultivars ICSV 400 and KSV 8 examined. The steep cycle incorporating 3 h length of air rest was found by experiment to give the optimum levels of malt quality indices analysed while at the same time causing considerable reduction in average main rootlet length and total malting loss for both grain cultivars.     

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 USE OF RAPID SCREENING TESTS TO COMPARE CHANGES DURING MALTING IN SORGHUM

Eight sorghum cultivars were malted and samples were taken after steeping and after each of 5 days germination. In genotypes with low levels of grain nitrogen, up to 35% of the nitrogen solubilisation during the malting process occurred in the steeping phase. After 5 days germination, extracts ranged from very low to moderate levels and correlated highly with diastase and half-grain mash results. In addition, genotypes producing higher extracts showed greater reductions in milling energy between 2 and 5 days germination. During malting, there was considerable genotype X day interaction for several quality characters, suggesting variation in rates of development as well as final quantities.     

Carbohydrate content and structure during malting and brewing: a mass balance study

A holistic view of the fate of barley starch, arabinoxylan and β-glucan throughout malting and brewing is largely missing. Here, an industrial scale malting trial and pilot brewing trial were performed, and the concentration and structural characteristics of carbohydrates were analysed at 28 key points in the process. The barley starch content decreased during malting from 75.0% to 69.7%. During mashing, malt starch was converted to fermentable sugars (75.3%), dextrin (22.8%) or was retained in spent grains (1.8%). Arabinoxylan was partially hydrolysed during malting. Despite mashing-in at 45°C, no further solubilisation of arabinoxylan was observed during mashing. However, the average degree of polymerisation of the soluble arabinoxylan fraction decreased slightly. During fermentation, the arabinoxylan content decreased to 2.5 g/L. The amount of barley β-glucan decreased gradually in time during malting. Of the solubilised β-glucan, 31% was retained in the spent grains during wort filtration, slightly lowering the β-glucan content in the wort. The β-glucan content remained at 0.5 g/L during fermentation. Sucrose was hydrolysed during mashing, probably by barley invertases. From the total amount of malt used, 41.0% was converted to fermentable sugars. This mashing yield could have been improved by the full hydrolysis to fermentable sugars of the present β-glucan (to 41.1%), the remaining starch in spent grains (to 42.0%) and dextrin in wort (to 50.3%). These results provide more insight into the carbohydrate conversions during malting and brewing and can act as a baseline measurement for future work. © 2020 The Institute of Brewing & Distilling     

THE USE OF ACETIC ACID AND SULPHUR DIOXIDE TO LIMIT MALTING LOSSES

The effects of spraying acetic acid or solutions of sulphur dioxide and of steeping in solutions of acetic acid have been investigated in micromalting trials. Spraying germinating grain with acid solutions on the fourth day of germination significantly increased both hot and cold water extracts and total soluble nitrogen, while reducing rootlet growth and total malting loss. The percentage fermentability of wort derived from malt sprayed with a solution of acetic acid was less than that of controls and had a lower pH. Malt prepared from grain steeped in dilute acetic acid and sprayed with gibberellic acid on casting was ready for kilning in a shorter time with smaller malting loss and rootlet production, but with an elevated hot water extract, percentage fermentability and soluble nitrogen content. On this basis of the original dry weight of barley, increases in the yield of extract of the order of 5% over that obtained from control malts have been achieved.     

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.     

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.     

THE MOLECULAR STRUCTURE OF GELPROTEIN FROM BARLEY,ITS BEHAVIOUR IN WORT-FILTRATION AND ANALYSIS

Gelproteins from barley and malt play a major role in the lauter tun filtration. These proteins (called glutenin in wheat) are partly broken down during malting and then aggregate during mashing. These aggregates form a part of the upper layer of the spent grains (the ‘oberteig’). To obtain more insight into this mechanism the composition of gelproteins of different cultivars of barley were analysed using various electrophoretic techniques. From this work a model can be proposed in which the protein, like its counterpart in wheat consists of high molecular weight (HMW) subunits as a backbone and low molecular weight (LMW) subunits as side chains, joined by inter-and intra-molecular disulphide bonds. During malting the disulphide bonds are reduced and subunits are broken down by proteolysis. During mashing a complex is formed between residual gelprotein in the malt and the glutelins, which gives rise to an impenetrable layer on the spent grains. The quantity of gelprotein as analysed by high pressure liquid chromatography (HPLC) or densitometry of Sodium dodecyl sulphate polyacrylamide electrophoresis (SDS—PAGE) patterns of HMW subunits showed a negative relationship to the lauter tun filtration rate.     

FACTORS INFLUENCING THE HARDNESS (MILLING ENERGY) AND MALTING QUALITY OF BARLEY

The hardness of samples of 41 barley cultivars grown in Australia was determined by measuring milling energy. The milling energy was negatively correlated (r= ?0.49) with the starch and positively correlated (r=0.50) with the β-glucan content. The correlation with protein content was not significant. This suggests that a low starch content and a high β-glucan content may contribute to hardness but other factors may probably also be involved. The extent of modification of these barley samples measured by Calcofluor staining after steeping and 48 h of germination was correlated with the grain hardness (r= ?0.56). Factors contributing to grain hardness may limit the rate of endosperm modification during malting, indicating the value of selecting softer cultivars for malting.     

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.     

THE PROTEIN RESERVES OF THE BARLEY GRAIN AND THEIR DEGRADATION DURING MALTING AND BREWING

The hordeins of barley are the main components of the grain protein, comprising B (sulphur rich), C (sulphur poor) and D (high relative molecular mass) species. A gel fraction can be isolated that consists of the D and part of the B hordein. Other important components of the reserve protein are β-amylase, protein Z and enzyme inhibitors as well as endosperm cell wall protein and protein associated with the starch granules. Degradation of protein is mediated by peptidase enzymes, most important of which are the endopeptidases and the exopeptidases; the latter having specificity for the carboxy- or amino-terminal region of a polypeptide chain. A very high activity of carboxypeptidase develops in the germinating barley grain. Five carboxypeptidases have been identified in malt and three fully characterized. Several endopeptidases have been identified, which have optimum activity close to pH 5.0 (that of the starchy endosperm) and studies with active site inhibitors suggest that sulphydryl dependent species are the most important. However, only limited use has been made of hordein as a substrate in the measurement of peptidase activity in malt. Lack of a complete understanding on the peptidases and their regulation in the germinating barley is a major limiting factor in the knowledge on the mobilization of the protein reserves during malting and mashing. Both de novo synthesis and activation of the peptidases have been demonstrated and the hormones gibberellic acid and abscisic acid appear to have a regulatory role. A controlled modification of the protein reserve is required so that there is an appropriate supply of amino acids for the brewers yeast but retention of proteins important in the desirable foaming component of beer. It is also important that proteins which can interfere in the final stages of brewing, e.g. those with chill haze forming potential, are broken down. An early hydrolysis is required of cell wall proteins and those proteins intimately associated with the starch granules.     

THE BREWING POTENTIAL OF “ACHA” (DIGITARIA EXILIS) MALT COMPARED WITH PEARL MILLET (PENNISETUM TYPHOIDES) MALTS AND SORGHUM (SORGHUM BICOLOR) MALTS

The malting and brewing characteristics of millets (Pennisetum typhoides and Digitaria exilis) and sorghum (Sorghum bicolor) were compared. Diastase, α-amylase, amyloglucosidase and proteases increased with malting time and the increase was associated with the modification. Development of hydrolytic enzymes was significantly higher in pearl millet and Digitaria exilis (“acha”) than in sorghum at P ≥ 0.01. The major starch degrading enzyme in the three varieties of pearl millet (SE composite, SE.13 and SE 2124) was α-amylase. On the other hand, β-amylase was the major starch degrading enzyme in “acha” (Digitaria exilis) which is similar to the pattern in barley. Gibberellic acid had a stimulating effect on the diastatic activity of pearl millets, Digitaria exilis (“acha”) and sorghum (KSV-4), but inhibited the diastatic activities of sorghum (Farafara). Gibbereltic acid inhibited the proteolytic activities in all the pearl millet varieties, Digitaria exilis and sorghum varieties. Potassium bromate had little or no effect in the reduction of malting losses. Although “acha” (Digitaria exilis) had a high β-amylase content, a high malting loss makes it uneconomical to brew with “acha” mart. A blend of “acha” malt with pearl millet malt or sorghum malt (composite malt) will produce a malt of the same profile as barley malt and this will enhance the quality of sorghum and pearl millet malt during the mashing process. Wort quality of all the samples was suitable for brewing conventional beer.     

THE COMPOSITION OF FINE PARTICLES WHICH AFFECT MASH FILTRATION

The fine particulate material from various types of mash has been analysed. These particles which influence the rate of wort separation consist of aggregates of protein, hemicellulose (pentosan and β-glucan), small starch granules and lipid. All grists investigated yielded such aggregates with similar qualitative composition but with marked variations in the relative amounts of individual components. There is no evidence that retrograded amylose contributes significantly to the composition of the aggregated particles. When heat stable β-glucanase is added to a mash containing 20% raw barley, neither the volume nor the β-glucan content of the fines changes appreciably but the texture is altered. The action of the glucanase loosens the structure of the particles to release small starch granules into the mash and the residual structure has a more porous appearance which probably allows freer passage of wort through the grain bed. The origin of mash-bed particles is the endosperm of the ungerminated barley or the under-modified regions of the malt.     

SOME RELATIONSHIPS BETWEEN THE PROTEIN NITROGEN OF BARLEY AND THE PRODUCTION OF AMYLOLYTIC ENZYMES DURING MALTING

Barleys studied (Chariot and Delibes) contained different levels of extractable β-amylase enzymes. The potential levels of β-amylase enzymes of the two varieties studied were similar at 1.4 to 1.7% total nitrogen. Higher values of potential β-amylase enzyme were observed in the Delibes barley of higher total nitrogen of 1.9%. The higher level of β-amylase found in the barleys with the highest total nitrogen was not reflected in the protein banding patterns as revealed by SDS-PAGE protein fractionation. Extraction of barley proteins was largely influenced by the different extractants used. The alcohol soluble proteins, M_r 97 kDa (D-hordeins), were only extracted when mercaptoethanol was included in the extracting solution. Although barleys with the highest nitrogen (1.9%) had the highest apparent potential to develop β-amylase enzymes, the better modified low nitrogen barleys produced higher levels of β-amylase and α-amylase when malted. Dehusking revealed that the high nitrogen barleys contained more steely grains. In contrast, the low nitrogen barleys contained more mealy grains. Steely grains contained more nitrogen than mealy grains and had the greater potential to develop β-amylase. Notwithstanding, the results of this study suggested that the proteins of the lower nitrogen barleys (1.4–1.7%) were capable of producing higher levels of β-amylase and α-amylase than the higher nitrogen barleys (1.9%) over comparable periods of malting. The high apparent β-amylase potential of the barley was linked to high nitrogen levels and associated high levels of steeliness, whilst the corresponding high β-amylase levels of malt were linked to the efficiency of endosperm modification of the malted grain.     

The (1–3, 1–4)‐β‐Glucanases in Malting Barley: Enzyme Survival and Genetic and Environmental Effects

Eighteen barley genotypes used in Brazilian malting barley breeding programs were characterized in relation to (1–3, 1–4)‐β‐glucanase activity in green and kilned malt. They were tested to determine the loss of enzyme activity during kilning in the malting process and the environmental effects on enzyme activity were measured. The genotypes analyzed showed great variation regarding the enzyme activity in both kinds of malt, in a range from 531.94 to 934.31 U/kg in green malt, and from 187.02 to 518.40 U/kg in dry malt. The mean enzyme activity loss during kilning was close to 60%, very similar to the results obtained in other studies. The loss among genotypes varied from 8.04% to 71.54%. The enzyme activity varied significantly under the different environments tested, showing existence of environmental effects on the genotypes analyzed. Embrapa 127 was the genotype that exhibited the highest enzyme activity in finished malt although it had shown a low activity in green malt, reflecting a negligible loss of activity during kilning. The data indicate promising results to malting barley breeding due to the wide variability exhibited by genotypes as to enzyme activity and levels of isoenzyme with high thermostability.     

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.     

A RAPID GUIDE TO ENDOSPERM MALTING POTENTIAL OF BARLEYS USING A SEDIMENTATION PROCEDURE

A rapid (10 minute) procedure is described which appears to differentiate barleys having good potential for malting by traditional methods from those less easy to malt. During controlled milling, barleys of good malting potential release more starch granules and, as a result, a suspension of the milled grain sediments more slowly in 70% ethanol. This probably reflects differences in the nature of the endosperm cell wall and the starch-protein matrix.