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.     

NON-STARCH POLYSACCHARIDES FROM BARLEY: STRUCTURAL FEATURES AND BREAKDOWN DURING MALTING

An extraction procedure is described for the complete solubilisation and fractionation of non-starch polysaccharides (NSP) from barley and malt, using ethanol, water and 4-methylmorpholine-N-oxide. Important structural features of the NSP are preserved with this non-destructive procedure. The fractionation method enables the investigation of structural features of NSP fractions and of NSP breakdown during malting.     

LIPOLYTIC AND OXIDATIVE CHANGES OF BARLEY LIPIDS DURING MALTING AND MASHING

Lipolytic and oxidative changes of barley lipids were studied during malting and mashing. The amount of lipid decreased by 23% during malting and changes in the composition of lipid classes were minor. On the other hand, during mashing the amount of free fatty acids (FFA) increased which indicated, that lipid hydrolysis had occurred. The same phenomenon was seen when malt flour was soaked in water at 23°C. The triglyceride (TG) and polar lipid (PL) contents were reduced and the proportion of FFA in total lipids was increased. Following similar soaking of barley flour, TG and PL were reduced but the accumulation of FFA and especially linoleic acid (LA) was slight. The results were consistent with the data on lipoxygenase activity (LOX) during malting. During steeping LOX decreased and was 15–20% of the activity of raw barley at the beginning of germination. The activity remained low during germination but rose sharply in the middle of kilning only to decrease again to a very low level at the end of kilning (5%). This in combination with the fact that the proportion of FFA remained high in the soaked malt samples suggests that the oxidation by LOX is negligible in the malt samples. However, the data suggest that mashing of barley, but not that of malt, includes the potential for the formation of highly polar lipid oxidation products.     

CHANGES IN BARLEY ENDOSPERMS DURING EARLY STAGES OF GERMINATION*

Kernels of 4 barley cultivars were germinated at 18°C and samples were removed for analysis at short time intervals for the first 30 h and at longer intervals during the ensuing 90 h. α-Amylase, (1 → 3) (1 → 4)-β-glucanase and (1 → 3) β-glucanase activities were measured in each sample. Analysis of kernel sections stained with Calcofluor showed that hydrolysis of β-glucan in the crushed cell layer commenced 6–9 h after the initiation of germination. Hydrolysis proceeded from the ventral edge to the dorsal edge of the kernel. Starch granule hydrolysis followed a similar pattern in the endosperm region adjacent to the crushed cell layer, but starch hydrolysis was always preceded by β-glucan hydrolysis.     

THE DEGRADATION OF β-GLUCAN DURING MALTING AND MASHING: THE ROLE OF β-GLUCANASE

Significant β-glucanolysis takes place during mashing and is catalysed by a β-glucanase which is specific to mixed-linkage β-glucans. The enzyme develops during the germination of barley, but is rapidly and extensively destroyed in kilning. Partially-purified preparations of β-glucanase are protected from denaturation by heat if their solutions are adjusted to pH 4 or if bovine serum albumin is added. However the most effective stabiliser of the enzyme is reduced glutathione. Oligosaccharides containing three and four glucosyl units are produced by the action of β-glucanase and they are further converted during malting and mashing by a different enzyme(s) to disaccharides and glucose.     

THE RELATIONSHIP BETWEEN β-GLUCAN SOLUBILASE,BARLEY AUTOLYSIS AND MALTING POTENTIAL

There is a correlation between the autolysis of barleys and their β-glucan solubilase activities. There is no correlation between autolysis and nitrogen content, β-glucan level, Milling Energy or Zeleny sedimentation value of the barley. Activities of endo-β-glucanase are inversely related to coarse-grind Hot Water Extract obtained from malts grown for 4 days. Whilst β-glucanase is not involved in the early stages of autolysis, β-glucan solubilase, present in large amounts in untreated barley, has a role both in extracting and degrading β-glucan. Barleys with low β-glucan content or high β-glucan solubilase modify more rapidly.     

THE RELATIONSHIP BETWEEN BARLEY EXTRACT VISCOSITY CURVES AND MALTING ABILITY

Twelve barley varieties, representing a wide range of malting ability, were examined using the Falling Time technique to determine their raw grain extract viscosities following extraction times ranging from 1 to 6 h. The shape of the viscosity curve was characteristic of each variety and was related to malting ability. A simpler index of potential malting ability was produced by the difference in Falling Time between extraction times of 3 and 4 h. This difference correlated well with the percentage extract obtained after micro-malting, (r = + 0·81, n = 31), and offers a possible method of assessing potential malting ability in barley breeding programmes.     

CHANGES IN THE CONTENTS OP BARLEY PROTEOLYTIC INHIBITORS DURING MALTING AND MASHING

Barley contains three types of large-molecular proteolytic inhibitors, viz., inhibitors active against malt endopeptidases, a specific trypsin inhibitor, and a group of inhibitors of microbial proteinases and chymotrypsin. The inhibitors of malt endopeptidases disappear at an early stage of germination, just before the endopeptidase activity begins to increase. The total amount of these inhibitors is so small, however, that the great increase in endopeptidase activity occurring during germination cannot be due to the breakdown of an “equivalent” amount of inhibitors. The contents of the other two types of inhibitor in barley are much higher; they decrease slowly during germination, but high activities are still present in malt. The inhibitors of microbial proteinases are progressively inactivated during mashing. The trypsin inhibitor retains full activity through mashing, but is completely inactivated during boiling and hopping.     

STUDIES ONα-GLUCOSIDASE IN BARLEY AND MALT*

A specific and sensitive method was used to determine α-glucosidase activity in barley and malt. Reliable results were obtained only after extracts of barley and malt had been dialyzed extensively to remove low molecular weight carbohydrates that interfered with the enzyme assay, α-Glucosidase was present in immature kernels of Bonanza and Ellice barley shortly after anthesis but enzyme levels fell rapidly as the kernels matured. A high proportion of the activity was present in pericarp tissue. Enzyme activity increased rapidly in Bonanza and Klages barley during initial stages of germination and fell only slightly during kilning. A high proportion of enzyme activity was present in the scutellum of 4-day germinated barley with lesser amounts in the aleurone and endosperm.     

THE MAJOR BIOCHEMICAL CONSTITUENTS OF AN ACID EXTRACT OF BARLEY FLOUR AS USED IN VISCOSITY DETERMINATIONS BY THE FALLING BALL METHOD

The viscosity of an acid extract of barley flour, measured as its ‘falling time’, has been used as a guide to malting potential.²² One such extract, made from a sample of Maris Mink, was examined in detail to identify the principle biochemical constituents and to investigate their contribution to viscosity. The extract contained a high concentration of carbohydrate (3·30 mg/ml) divided into two discrete fractions of high and low molecular weight (in the weight ratio of 39:61) by gel filtration. The high molecular weight component was polydisperse and consisted principally of β-glucan with some starch and pentosan also present. The low molecular weight fraction contained mostly fructose, glucose and some pentose. Protein present in the extract (1·07 mg/ml) was derived almost exclusively from the albumin and globulin (salt soluble) fraction of barley protein. Studies on the viscosities of reconstituted preparations of these components indicated that β-glucan made the major contribution to the viscosity of the extract. This conclusion is supported by observed changes in viscosity after the extract was treated with β-glucanase, α-amylase and protease.     

EFFECTS OF CULTIVAR AND ENVIRONMENT ON β-GLUCAN CONTENT AND MALTING QUALITY OF TURKISH BARLEYS

Eight barley cultivars grown under the same agronomic conditions and samples of Tokak cultivar grown at six different sites of Turkey were used in this study. There were significant differences among the barley cultivars and growing locations in terms of β-glucan content (p<0.05). Among malt quality criteria tested for the 8 barley cultivars; friability, viscosity, Kolbach index and extract difference showed significant correlations (p<0.05) with the total β-glucan content. Similar correlations were also observed between the malt quality criteria (Kolbach index and extract difference) and β-glucan contents for the Tokak samples grown at different sites.     

EVALUATION OF MALTING QUALITY IN A BARLEY BREEDING PROGRAMME USE OF α-AMYLASE AND β-GLUCAN LEVLES IN MALT AS PRESELECTION TOOLS

Analysis according to the EBC protocol, immunological determination of a α-amylase and estimation of malt β-glucan using the Calcofluor-FIA method, were used to screen 327 barley breeding lines for malting quality. The results obtained with the α-amylase and β-glucan methods are highly correlated to the important malt quality paramters: extract yield and β-glucan content in the wort. It is recommended that either of the two methods, which are simple to perform are used as prescreening tools in breeding programmes for malting barley.     

A ROLE FOR CARBOXYPEPTIDASE IN THE SOLUBILIZATION OF BARLEY β-GLUCAN

An enzyme is described which catalyzes the release of soluble β-glucan from insoluble barley endosperm cell walls. This enzyme increases in activity throughout malting. It has been partially purified and found to behave in the same way as an acidic carboxypeptidase on isoelectric focusing and in its sensitivity to inhibitors and activators and to heating. The importance of the β-glucan solubilizing enzyme in malting and mashing is discussed. An improved method for β-glucan determination is described.     

AN APPROACH TO THE ESTIMATION OF BREWING QUALITY IN BARLEY AND MALT

Methods of assessing the brewing qualities of new varieties are reviewed. A new method for making this assessment is proposed. This is ideally a three-step process but it would be possible to eliminate the third, costly, step without severe loss of effectiveness. The use of the new method is illustrated by reference to trials using 251 samples from recent trials in France.     

THE CAMBRIDGE PRIZE LECTURE 1995: INDIRECT AND DIRECT CONTRIBUTIONS OF BARLEY MALT TO BREWING

The suitability of barley malt as a raw material for brewing is determined by an amalgamation of “indirect” and “direct” contributions to the beer produced. Indirect contributions are considered as those which affect the quality of the brewing process performance whereas direct contributions are considered as those which affect the quality of the product. As a potential indirect contribution of malt to brewing quality evidence is presented that barley malt contains a flocculent which influences mash filterability. As a potential direct contribution of barley malt to beer quality evidence is presented that the mineral silicate found in beer may have a role in moderating dietary aluminium.     

‘FREE’ AND ‘BOUND’ β-AMYLASES DURING MALTING OF BARLEY. CHARACTERIZATION BY TWO-DIMENSIONAL IMMUNOELECTROPHORESIS

Major qualitative and quantitative changes in the β-amylases and in other salt soluble barley proteins occurred during the first four days of germination. Two soluble forms of barley β-amylase, ‘free’ β-amylase and β-amylase aggregated with a non-active protein Z, were found in extracts from all stages. A third enzyme form appeared during malting. Immunoelectrophoretic characterization seemed to support the possibility that this enzyme form could be a product of ‘bound’ β-amylase solubilization. All soluble forms of β-amylase and of protein Z in malt were electrophoretically heterogeneous. Two different, immunochemically related forms of protein Z present after malting retained their immunoelectrophoretic properties during brewing and were found to be dominant antigens in beer.     

MOBILISATION OF ENDOSPERMAL RESERVES DURING THE GERMINATION OF BARLEY

The metabolism of the germinating barley seed is a complicated physiological process. From the point of view of industrial malting, the syntheses of amylolytic and proteolytic enzymes as well as the degradation of endospermal structures are extremely important. In order to control his process the maltster needs to have a good understanding of these processes as well as of the regulatory mechanisms which control them. When a good malting barley is allowed to germinate under traditional conditions for the time needed, a good malt can be produced without this scientific background. The present day brewing industry, however, has to work with barleys which are not of the traditional malting type and furthermore accelerated processes are an economic necessity. Under these circumstances, the maltster needs all scientific help he can get. Biochemical and physiological research on germination provides him with the tools he needs for process control and optimisation.     

APPLICATION OF A COMMERCIAL ENZYME PREPARATION IN THE BARLEY MALTING PROCESS

This paper presents data on barley micromalting with addition of the CELLUCLAST enzyme complex. This is a commercial, multicarbohydrase with distinct β-glucanase and proteinase activities. The enzyme was added to steeping and germination phases, in different quantities (0.05%; 0.75% and 0.1% of initial barley). The enzyme was added to different malting phases: to the 2nd and 3rd steep water, at the beginning of germination on the 1st day by spraying, on the 2nd day of germination and in combination of addition to 3rd steeping water and in germination start (50% of total quantity of each). CELLUCLAST enzyme had a significant effect on reduction of wort viscosity, extract difference, wort filterability and protein breakdown, depending on the quantity of added enzyme and the malting phase to which it was added. There was no negative effect on other malt quality parameters. The best values of cytolytic breakdown parameters (viscosity, extract difference, filtration rate) were obtained with addition of 0.075% of CELLUCLAST, on the first day of germination.     

THE DETERMINATION OF GERMINATED GRAINS IN BARLEY

A method for determining germinated grains in barley by detecting the presence of α-amylase activity in individual corns is recommended.     

AN APPROACH TO THE IDENTIFICATION OF A ß-GLUCAN SOLUBILASE FROM BARLEY1

A ß-glucan solubilase activity was demonstrated in barley extract. This enzyme catalyzed the dissolution of barley ß-glucan by releasing a product having a narrow molecular weight distribution and a molecular weight of about 20,000. The enzyme was partially purified by ion exchange chromatography on DEAE-cellulose and gel permeation chromatography on Bio-Gel P-100. Although carboxypeptidase activity was present in the crude extract of barley flour the partially purified ß-glucan solubilase did not hydrolyse N-CBZ-Phe-ala. Examination of extracts from different barley tissues indicated that the ß-glucan solubilase activity was associated with the husks only; a large portion of the activity was extractable from whole barley kernels. About 85% of the enzyme activity in crude extracts from barley flour was retained after 40 min at 62°C. However, the enzyme was much more heat-labile in extracts of whole barley kernels. The pH of maximal activity was found to be about pH 5.7 and results from column chromatography suggested that the enzyme had a low pl value and a MW between 5 × 10⁴ and 6 × 10⁴.