Improvement of malt modification by use of Rhizopus VII as starter culture

The development of a selected starter culture on malting barley and its effects on malt quality aspects were studied. Application of Rhizopus sporangiospores in a malting process resulted in increased beta-glucanase and xylanase contents of the malting barley and improved starchy endosperm cell-wall degradation. Activation of the sporangiospores and optimization of the inoculation procedure led to a further increase in enzyme levels and to larger and more consistent impacts on cell-wall modification. Whereas the main effect of the starter culture on beta-glucan degradation was observed during malting, a further decrease in beta-glucan during mashing suggests that the microbial enzymes that survived the kilning step were active during mashing. Other quality aspects that were influenced by the starter culture activity were protein modification, wort color, and wort pH. The level of microbial enzymes produced was related to the amount of barley kernels infected with the starter culture.     

The effect of mashing on malt endoproteolytic activities

During malting and mashing, the proteinases of barley (Hordeum vulgare L.) and malt partially hydrolyze their storage proteins. These enzymes are critical because several aspects of the brewing process are affected by the soluble proteins, peptides and/or amino acids that they release. To develop improved malting barleys and/or malting and brewing methods, it is imperative to know whether and when the green malt endoproteinases are inactivated during malting and mashing. These enzyme activities are totally preserved during kilning and, in this study, we have determined when they were inactivated during mashing. Samples were removed from experimental mashes that mirrored those used in commercial breweries and their endoproteolytic activities were analyzed. The malt endoproteinases were stable through the 38 degrees C protein rest phase, but were quickly inactivated when the mash temperature was raised to 72 degrees C for the conversion step. All of the proteinase activities were inactivated at about the same rate. These findings indicate that the soluble protein levels of worts can be varied by adjusting the protein rest phase of mashing, but not by altering the conversion time. The rates of hydrolysis of individual malt proteins probably cannot be changed by altering the mash temperature schedule, since the main enzymes that solubilize these proteins are affected similarly by temperature.     

Image Analysis of Grain and Chemical Composition of the Barley Plant as Predictors of Malting Quality in Mediterranean Environments

We have explored the possibility of predicting the malting quality of barley grain, indicated by malt extract yield, by characteristics measured either on plants at anthesis or in mature dry grain by image analysis. To produce barley samples with varying levels of all the characteristics studied, we used grain from an experiment designed to study the influence of lowinput husbandry practices on malting quality of barley by growing five malting genotypes at each of four environments (site × season) and with two different agronomic treatments (N fertilization and herbicide-mechanical roguing of weeds). The results showed that nitrogen content in the plant at anthesis was a good predictor of grain protein content, this characteristic in turn being positively correlated with embryo size and grain volume, as estimated by image analysis, and negatively correlated with nonstructural carbohydrate content in the plant at anthesis. Extract yield was positively correlated with Kolbach index (ratio of soluble to total wort protein) and negatively correlated with wort viscosity and barley grain protein content. Thus, the only practical predictor of malt extract was grain protein content.     

Cropping sequence and nitrogen fertilizer effects on the productivity and quality of malting barley and soil fertility in the Ethiopian highlands

The productivity and quality of malting barley were evaluated using factorial combinations of four preceding crops (faba bean, field pea, rapeseed, and barley) as main plots and four nitrogen fertilizer rates (0, 18, 36, and 54 kg N ha^?1) as sub-plots with three replications at two sites on Nitisols of the Ethiopian highlands in 2010 and 2011 cropping seasons. Preceding crops other than barley and N fertilizer significantly improved yield and quality of malting barley. The highest grain yield, kernel plumpness, protein content, and sieve test were obtained for malting barley grown after faba bean, followed by rapeseed and field pea. Nitrogen fertilizer significantly increased yield, protein content, and sieve test of malting barley. All protein contents were within the acceptable range for malting quality. Inclusion of legumes in the rotation also improved soil fertility through increases in soil carbon and nitrogen content. We conclude that to maximize yield and quality of malting barley, it is critical to consider the preceding crop and soil nitrogen status. Use of appropriate break crops may substitute or reduce the amount of mineral N fertilizer required for the production of malting barley at least for one season without affecting its quality.     

Hardness Changes and Endosperm Modification During Sorghum Malting in Grains of Varying Hardness and Malt Quality

This study examined the interaction between sorghum grain hardness and sorghum malt quality in terms of diastatic power and free amino nitrogen with endosperm modification during malting. The changes in kernel hardness during malting of four commercial sorghum cultivars of differing quality in terms of endosperm texture and potential malt quality were measured using tests for hardness and density, and endosperm modification was followed by scanning electron microscopy. The general pattern of modification during sorghum malting was confirmed to start at the endosperm–scutellum interface and then continue into the floury endosperm toward the kernel distal end. Significantly, a cultivar of intermediate hardness and low malting quality remained harder and modified more slowly than a harder cultivar of high malting quality. It appeared that intrinsic grain hardness and malt amylase and protease activity both affected malt hardness and endosperm modification, but amylase and protease activity had a greater effect because of their degradation of endosperm starch and protein. Of the hardness and density tests studied, the tangential abrasive dehulling device (TADD) gave the best measure of hardness throughout malting; maximum range was 24–100% kernel removed over five days of malting. Also, the data agreed with the observed malt modification rates. Thus, the TADD may have application as a simple and rapid test for estimating sorghum malt quality.     

Degradation of starchy endosperm cell walls in nongerminating sterilized barley by fungi

Strains of fungi from different origins, including isolates of the natural microflora of barley, were screened for their ability to modify barley starchy endosperm cell walls in situ. In an initial step, fungi were selected that degrade the major component of the cell walls, that is, (1-->3),(1-->4)-beta-D-glucan, in vitro on artificial media. Nongerminating, sterilized barley, obtained by gamma-irradiation, was inoculated with such fungi and subjected to solid state fermentation under conditions resembling those of a traditional malting process. For some strains of fungi, a clear correlation between the production of endo-beta-glucanase and the friability of the treated kernels was found. Image analysis of Calcofluor stained longitudinal sections of barley kernels fermented with the endo-beta-glucanase producing strains showed that starchy endosperm cell walls were modified. As malt quality is inversely related to its (1-->3),(1-->4)-beta-D-glucan content, the selected strains have high potential to be used as starter cultures during malt production, contributing to the processing quality of the final product.     

Effect of Broken Corn Levels on Water Absorption and Steepwater Characteristics

Broken corn created by grounding sound corn kernels was added back at levels of 0, 4, 8, 12, or 16%, by weight, to whole kernels of three corresponding hybrids: FR27 × FRMo17 (a soft endosperm corn), FR618 × FR600 (amedium‐hard endosperm corn), and FR618 × LH123 (a hard endosperm corn). The samples had been dried from 28% moisture content to 15% moisture content either by using ambient air at ≈25°C or at 110°C. Samples were steeped for 36 hr at 52°C in 0.15% sulfur dioxide and 0.5% lactic acid steeping solution. The steepwater characteristics, such as water absorption, solids and protein content in the steepwater, and steepwater pH, were measured by periodic sampling and analyzed. Broken corn level has a significant effect on the amount of solids released during steeping and steepwater protein content for all samples. Both steepwater solids and protein content increased linearly as broken corn content increased. Corn drying temperature, kernel hardness, and interactions between drying temperature and kernel hardness has a significant effect on steepwater solids and protein content and steepwater pH in both broken and unbroken corn. Corn dried at low temperature released more soluble solids and protein into the steepwater than corn dried at high temperature. Soft endosperm and medium‐hard endosperm corn released more soluble solids and protein into the steepwater than hard endosperm corn. Soft endosperm corn resulted in a higher steepwater pH than medium‐hard and hard endosperm corn. No significant effect of broken corn content on final moisture content of steeped corn and steepwater pH was observed.     

Barley viability during storage: use of magnetic resonance as a potential tool to study viability loss

Malting-quality barley samples of the varieties Harrington, Manley, and TR118, each from two locations in Saskatchewan, were collected directly from the producers and sent to China for storage. At regular intervals samples were shipped back to Canada for analysis consisting of germination studies, alpha-amylase tests, magnetic resonance imaging (MRI), and magnetic relaxation (NMR) studies. Samples showing a decrease in germinative energy and elevated levels of alpha-amylase also showed a rapid uptake of water in the area between the embryo and the endosperm as observed by MRI. Using NMR relaxation experiments, viable and nonviable barley samples could be distinguished after 2 h of imbibition.     

Germination and Malting Properties of Mutants Derived from Malting Barley cv. Triumph

Four mutants, demonstrating a range of dormancy, were derived from the malting barley cv. Triumph. Although there were environmental effects on the rate of recovery from dormancy, relative performance of the genotypes was consistent. Recovery from water sensitivity was slower than recovery from dormancy for all genotypes, but a similar ranking of genotypes was observed with two mutants germinating more readily than the parental genotype. Exposure of the grain to the plant hormone abscisic acid (ABA) at the end of each wet phase during steeping had a highly significant effect on the malting performance of all samples. However, reduction in extract levels was significantly less in the two mutants that demonstrated more rapid recovery from dormancy. None of the mutants exceeded Triumph for hot water extract level after malting in two seasons at sites in Dundee (eastern Scotland) and Lleida (northeastern Spain). However, one mutant combined rapid recovery from dormancy with high extract levels when grown and malted under Scottish conditions and subjected to unithermal hot water extraction.     

Esterases in Barley and Malt

Several esterases from barley and malt have been separated on polyacrylamide gels. The slowest moving bands appear to represent a single enzyme displaying a spread of migration owing to differences in surface charge. During malting, this enzyme, which is located in the starchy endosperm, shifts to a more migratory form. Two other main esterase groups are identified through gel electrophoresis, notably a highly anionic, highly labile enzyme, MW 62,000, located in the aleurone. The slowest and fastest moving bands have been partially purified using salt fractionation and ion‐exchange chromatography. The former, MW 47,000, has strong capability for hydrolyzing acetylxylan and it is speculated that its role in the starchy endosperm is as part of the enzyme system that hydrolyzes the cell walls.     

Phosphorus seed coating as starter fertilization for spring malting barley

Abstract

Experiments were conducted with malting barley (Hordeum vulgare L.) cultivars in 2003 and 2004 in central, western and southern Lithuania to test phosphorus (P) availability at early seedling growth stage with P seed coating. Phosphorus seed coating resulted in alteration to plant stand structure traits. Despite the fact that seedling emergence sometimes decreased, the number of total and productive stems, number of grains per ear and 1000-grain weight increased. The positive effect of P seed coating on grain yield was revealed when the growth conditions during post-anthesis were favourable for exploiting the potential that was obtained during the pre-anthesis phase. In our experiment favourable conditions were considered those that generated a grain yield over 6 t ha^?1. According to path coefficients analyses the significant positive effect of P seed coating on malting barley yield increase was related to seed weight increase. Although P seed coating slightly increased single grain N concentration and thus grain protein concentration, grain protein concentration ranged from 8.6% to 10.2% and met the quality standard requirements for malting barley in all trials and treatments.     

REVIEW: Oat and Rye β‐Glucan: Properties and Function

Over the years, the β‐glucan of oats and barley has been the subject of study either because of the importance of the cholesterol‐lowering potential to health claims (FDA 1997, 2005) or, in the case of barley, because of the role of β‐glucan and β‐glucan‐rich endosperm cell walls in malting and brewing. β‐Glucan is also present in rye and in much lesser amounts in wheat. The most striking difference in these latter two sources is the difficulty in extractability; alkali rather than water is required for significant release from the cell walls. This review will discuss physicochemical properties of oat and rye β‐glucan and, where information allows, relate these to physiological effects. Viscosity, or more generally rheology, plays a central role in discussions of cereal β‐glucan functionality and physiological effects and will be the focus of this review.     

Influence of Barley and Malt Storage on Lipoxygenase Reaction

The dioxygenation of linoleic acid (LA) by aqueous flour suspensions of barley and malting samples was studied. The rate of this lipoxygenase (LOX) reaction varied as the malting process proceeded, giving a characteristic LOX reaction profile for a malting. The differences in the profiles from one malting to another were dramatic. It also appeared that during storage of dry, intact kernel samples from a single malting, a reduction in the rate of LOX reaction always occurred, and the rates of reduction with time were dependent on the stage of malting at the time of sampling. The kinetics of this aging could roughly be divided into four categories representing different stages of malting. Consequently, greatly varying LOX reaction profiles can be obtained from a single malting depending on the time of storage of kernels before assays. The results indicate that steeping, germination and the subsequent drying render the state of kernels unstable with respect to the LOX reaction for at least two to three weeks. Homogeneity of malt quality is important in the further applications of malt, especially in the brewing industry. Therefore, the rate of LOX reaction should be considered as a quality factor of malt.     

Release of β‐Glucan from Cell Walls of Starchy Endosperm of Barley

β‐Glucan can be solubilized from barley by warm water, with increasing solubilization as the temperature is increased. Substantially less glucan is extracted if the barley is dehusked using sulfuric acid, particularly if the dehusked barley is denatured. This indicates that enzymes capable of solubilizing glucan are present in barley. Various purified enzymes promote the solubilization of glucan from denatured and dehusked barley. Apart from endo‐β‐(1→3)(1→4)‐glucanase, these enzymes include endo‐xylanases, arabinofuranosidase, xyloacetylesterase, and feruloyl esterase. Ferulic acid and, probably, acetyl groups are esterlinked to arabinoxylan, not β‐glucan, in the cell walls of barley starchy endosperm, so the ability of the esterases, xylanases, and arabinofuranosidase to solubilize glucan indicates the pentosan component of the cell wall can restrict the extraction of glucan.     

Chlorophyll meter as nitrogen management tool in malting barley

Abstract

Variable precipitation in many regions makes it difficult to predict yield goals and nitrogen (N) rates for malting grade barley (Hordeum vulgare L.). During years with below normal growing season precipitation, barley fertilized at the recommended rate often exhibits grain protein concentrations exceeding what is acceptable for malting. A study was conducted to evaluate the chlorophyll meter as a N management tool. Barley was grown under several N rates in the field. Chlorophyll meter readings and N additions were made at the Haun 4 to 5 growth stage, and grain yield and protein concentrations were evaluated at maturity. Chlorophyll meter readings, normalized as meter reading from treatment plot divided by that from a plot receiving a full N treatment at the Haun 4 to 5 growth stage, were correlated with grain yield (r²=0.67). Stands having normalized chlorophyll meter readings below 95% responded to N additions with yields equivalent to the fully fertilized stand and grain protein concentrations acceptable for malting. A N management strategy is proposed whereby 40 to 50% of the N calculated for the yield goal is applied at planting and a fully fertilized reference strip is included for each variety or soil type. At the Haun 4 to 5 growth stage, chlorophyll meter readings are taken in the reference strip and in the field. Normalized chlorophyll meter readings below 95% of the reference strip indicate a need for additional N fertilizer. This strategy will provide producers with additional time (up to a month) to evaluate growing season conditions before investing in additional crop inputs and will improve the likelihood that a barley crop acceptable for malting will be produced.     

Impact of low hydration of barley grain on β-glucan degradation and lipid transfer protein (LTP1) modifications during the malting process

One of the objectives of the malting industry is to reduce the energy cost during kilning without major effect on malt quality. In this study, the impact of a low hydration steeping process on lipid transfer protein (LTP1) modifications and β-glucan breakdown was evaluated in low (LH) and high (HH) hydrated malts. LTP1 modifications analyzed by MS/MS revealed acylation, glycation, and disulfide bond breakage in both LH and HH malts. LTP1 free amine content measurement and fluorescence of Maillard protein adducts revealed no significant difference between LH and HH malts. Immunolabeling of LTP1 during malting highlighted the diffusion of the protein from the aleurone layer to the endosperm at the end of steeping in both LH and HH malts. By contrast, a significant higher amount of β-glucans was measured in LH malts after five days of germination, whereas no significant difference between LH and HH malts was revealed through immunostaining of β-glucans or evaluation of the endosperm integrity after seven days of germination. The possibility to reduce the effects of a low hydration steeping process on β-glucan hydrolysis by increasing germination time was discussed.     

Malting Oats: Effects on Chemical Composition of Hull-less and Hulled Genotypes

Samples of hull-less oat genotypes from the Cooperative Naked Oat Test grown in Ottawa, ON, and Aberdeen, ID, were analyzed for their potential as a food malt. Malted oats had a lower concentration of petroleum ether-extractable lipid, but a much higher percentage of the lipid was in the form of free fatty acids. About 5% less starch and slightly more N was found in malted oats than in unmalted. Malted oats contained ≈8% soluble carbohydrate. During the germination phase of malting, nearly all the β-glucan was degraded. α-Amylase activity of malted oats approached that of malting barleys, but diastatic power was much lower. Groats of hulled cultivars grown at Madison, WI, were malted and analyzed with similar results. Because the increased levels of free fatty acids in the malted grains may lead to the development of rancid flavors, a method to curtail their increase or selections of genotypes with a minimum increase during malting may be necessary to produce a useful malted food product.     

Quantitative study of the formation of endoproteolytic activities during malting and their stabilities to kilning

The proteinases of germinating barley (Hordeum vulgare L.) hydrolyze storage proteins into amino acids and small peptides that can be used by the growing plant or, during brewing, by yeast. They are critical for the malting and brewing processes because several aspects of brewing are affected by the amounts of protein, peptide, and amino acids that are in the wort. This study was carried out to quantitatively measure when endoproteinases form in green malt and whether they are inactivated at the high temperatures that occur during malt kilning. Little endoproteolytic activity was present in ungerminated barley, but the activities began forming 1 day into the "germination" phase of malting, and they were nearly maximal by the third germination day. Quantitative studies with azogelatin "in solution" assays showed that the green malt endoproteolytic activities were not inactivated under commercial kilning conditions that use temperatures as high as 85 degrees C but that some actually increased during the final kilning step. Qualitative (2-D, IEF x PAGE) analyses, which allow the study of individual proteases, showed that some of the enzymes were affected by heating at 68 and 85 degrees C, during the final stages of kilning. These changes obviously did not, however, decrease the overall proteolytic activity.     

Estimating nitrogen requirements for irrigated malting barley

Abstract

The production of marketable malting barley requires careful N management to meet the quality standards set by the malting industry. Nine field trials were conducted over an eight‐year period at four locations to develop N fertilization guidelines for irrigated malting barley. Residual soil NO₃‐N (0 to 60 cm) ranged from 15 to 103 kg N/ha. Nitrogen fertilizer was applied preplant as either urea or NH₄NO₃ at rates ranging from 0 to 269 kg N/ha. Maximum yields were obtained when the sum of residual plus applied N (available N) was above 110 kg N/ha. However, the percentage of plump kernels generally fell below acceptable levels (85%) when available N exceeded 135 kg N/ha. Grain protein exceeded acceptable levels (12%) when available N was above 210 kg N/ha. Stem NO₃‐N sufficiency levels were determined from high‐yielding barley with acceptable quality parameters. At the three‐leaf stage, the barley stem NO₃‐N sufficiency level was approximately 6,000 μg/g and decreased to about 1,000 μg/g at the eight‐leaf stage.     

Mutation analysis of barley malt protein Z4 and protein Z7 on beer foam stability

Beer foam stability is an important characteristic. It has been suggested that isoforms of protein Z, that is, protein Z4 and protein Z7, contribute to beer foam stability. We investigated the relationship between beer foam stability and protein Z4 and protein Z7 using their deficient mutants. As a protein Z4-deficient mutant, cv. Pirkka was used. Protein Z7 deficiency was screened in 1564 barley accessions in the world collection of Okayama University, Japan. The barley samples from normal, protein Z4-deficient, protein Z7-deficient, and double-deficient were genotyped in F(2) populations and then pooled based on the DNA marker genotypes of protein Z4 and protein Z7. For a brewing trial, F(5) pooled subpopulations were used. After malting and brewing, the foam stability was determined, and the results showed that the levels of foam stability in the four samples were comparable. Two-dimensional gel electrophoresis was used to investigate the proteome in these beer samples. The results showed that low molecular weight proteins, including lipid transfer protein (LTP2), in the deficient mutants were higher than those in the normal sample. Our results suggest that the contribution of protein Z4 and protein Z7 to beer foam stability was not greater than that of other beer proteins.