Effect of malting time,mashing temperature and added commercial enzymes on extract recovery from a Nigerian malted yellow sorghum variety

Preliminary microbiological studies carried out on sorghum grains showed that the major microorganisms found were mainly bacteria and that aflatoxin‐producing fungi were not found. The effect of added commercial enzyme preparations and different infusion mashing temperatures on extract yield, from sorghum malted at 30 °C, was studied. The infusion mashing method (65 °C) developed for mashing well‐modified barley malt produces poor extract yields with sorghum malt. The extract yield from the sorghum malt in this study was very low with infusion mashing at 65 °C, without the addition of commercial enzyme preparations. A higher extract yield was obtained from the sorghum malt, without the commercial enzyme addition, when using infusion mashing at 85 °C. Both infusion mashing temperatures (65 and 85 °C) showed an improved extract yield over the control malt when commercial enzyme preparations were used during mashing of the sorghum malt. The added enzyme preparations resulted in a higher extract yield from the germinated sorghum when infusion mashing was performed at 65 °C over mashing at 85 °C. The use of individual commercial enzymes (α‐amylase, β‐glucanase, protease, xylanase, saccharifying enzyme and combinations of some hydrolytic enzyme) increased extract yields, when complemented with the enzymes that had developed in the sorghum malt. Copyright © 2016 The Institute of Brewing & Distilling     

DEVELOPMENT OF A MASHING PROFILE FOR THE USE OF MICROBIAL ENZYMES IN BREWING WITH RAW SORGHUM (80%) AND MALTED BARLEY OR SORGHUM MALT (20%)

Different time and temperature programmes were used to evaluate the production of hot water extract (HWE) and free amino nitrogen (FAN) from mashes containing raw sorghum and either malted sorghum or malted barley in the presence of microbial enzymes. Two malted varieties of sorghum (SK 5912 and Zaria) were used. The former gave higher HWE but lower FAN than the latter. Sorghum malts were unable to provide enzyme activity for starch extraction and exogenous enzymes were always needed. Seventeen commercially available enzyme preparations were assessed. A double-mash process was developed. Inclusion of calcium ions (200 ppm) was beneficial but adjustment of mash pH had little effect. Raw sorghum was gelatinised at 100°C for 30–40 min in the presence of a heat-stable α-amylase followed by mixing with a malt mash (started at B0°C) to give a temperature of 65°C with a total mash time of 167 min (127 min from mixing the mashes). The inclusion of a single commercial enzyme preparation (containing both proteolytic and amylolytic activities) was sufficient to achieve satisfactory HWE and FAN. Addition of different activities or combinations of activities gave no significant advantages. To obtain levels of FAN of 100–140 mg/l however excessive amounts of enzymes were required.     

α-GLUCOSIDASE ACTIVITY OF SORGHUM AND BARLEY MALTS

Sorghum malt α-glucosidase activity was highest at pH 3.75 while that of barley malt was highest at pH 4.6. At pH 5.4 employed in mashing sorghum malt α-glucosidase was more active than the corresponding enzyme of barley malt. α-Glucosidase was partly extracted in water but was readily extracted when L-cysteine was included in the extraction buffer, pH 8. Sorghum malt made at 30°C had higher α-glucosidase activities than the corresponding malts made at 20°C and 25°C. Nevertheless, the sorghum malts made at 20°C and 25°C produced worts which contained more glucose than worts of malt made at 30°C. Although barley malts contained more α-glucosidase activity than sorghum malts, the worts of barley had the lowest levels of glucose. The limitation to maltose production in sorghum worts, produced at 65°C, is due to inadequate gelatinization of starch and not to limitation to β-amylase and α-amylase activities. Gelatinization of the starch granules of sorghum malt in the decantation mashing procedure resulted in the production of sorghum worts which contained high levels of maltose, especially when sorghum malt was produced at 30°C. Although the β-amylase and α-amylase levels of barley malt was significantly higher than those of sorghum malted optimally at 30°C, sorghum worts contained higher levels of glucose and equivalent levels of maltose to those of barley malt. It would appear that the individual activities of α-glucosidase, α-amylase and β-amylase of sorghum malts or barley malts do not correlate with the sugar profile of the corresponding worts. In consequence, specifications for enzymes such as α-amylase and β-amylase in malt is best set at a range of values rather than as single values.     

Effect of malting regimen on diastatic power,cold and hot water extracts of malts from sorghum

In this study, sorghum grains were malted using different malting regimens and the malt qualities were analysed for cold water extract (CWE), hot water extract (HWE) and diastatic power (DP). Results showed that malting regimen produced sorghum malts with peak CWE, HWE and DP within the pre-determined limits of malting regimen. From this, predictive models of sorghum malt quality were developed and tested. Whilst steeping duration interacts significantly with the germination period to cause variations in CWE and HWE, only the germination period significantly influenced the DP of sorghum malts. However, the kilning temperature showed no significant impact on the parameters studied. Besides CWE, HWE and DP correlating positively with each other, the developed predictive models were significant (P < 0.05) and satisfy the quality of fit (R² > 0.70) of the proposed models. The result demonstrated that wort for different purposes can be obtained by varying the malting operational conditions in the predictive model, thus saving time, resources, preliminary studies and research designs. Besides the promotion of a cheaper local alternative material for industrial wort production and brewing purpose, this research provides valuable insight on the brilliant prospect of variable malting regimen in predicting and adjusting the quality characteristics of sorghum malt and wort. Future studies are required to develop models for wort produced from peak CWE, HWE and DP, for predicting wort quality characteristics.     

ENZYMIC BREAKDOWN OF ENDOSPERM PROTEINS OF SORGHUM AT DIFFERENT MALTING TEMPERATURES

Enzymic breakdown of endosperm proteins of sorghum was more effective at 20°C than at 25°C and 30°C, as regards total protein solubilization, α-amino nitrogen and peptide production. Although the embryos (axes and scutella), of the three temperature treatments contained similar quantities of protein, it appeared that less proteins, in terms of amino acids and peptides, were transferred to the roots during malting at 30°C than at 25°C and 20°C. During mashing, higher levels of peptides but lower levels of α-amino nitrogen and total soluble nitrogen were released in an infusion mash at 65°C than in a decantation mash where enzymically active wort was decanted and used to mash gelatinized sorghum starch at 65°C. Although more of the maltose-producing enzyme—β—amylase was found in sorghum malts made at 25°C and 30°C than at 20°C, it would seem that, for sorghum, malting temperature of 20°C to 25°C were optimal as regards protein breakdown during malting. The protein breakdown produced when sorghum is malted at 20°C is comparable to that found in barley malt and should support similar levels of adjuncts and yeast growth during brewing.     

CHANGES IN ACTIVITY OF SORGHUM LIPASE DURING MALTING AND MASHING

Lipase activity was monitored during malting and mashing of sorghum grains. All three sorghum varieties contained detectable lipase activity in the ungerminated form. Lipase activity changed only slightly during steeping for 24 hours but increased several fold in the course of germination. Between 24% and 60% of the lipase activity of the green malt was retained after kilning at 48°C but no activity was detected in the wort after mashing at 65°C. About 68% of the lipase activity of 72 hours old malt was detected in the plumule, while 29% and 3% were detected in the endosperm and radicle, respectively. Optimum activity was observed at pH 7.0.     

Mashing Studies with Unmalted Sorghum and Malted Barley

The effects on wort quality when mashing with unmalted sorghum (0–100%) and malted barley (100–0%) in combination with industrial enzymes were evaluated. A mashing program with temperature stands at 50°C, 95°C and 60°C was used. Different combinations of commercial enzymes were evaluated. A heat stable α‐amylase was found to be essential for efficient saccharification. The inclusion of a fungal α‐amylase in mashes with a high sorghum content improved filtration rates to that of 100% malted barley mashes. Addition of a bacterial protease increased the amount of nitrogen solubilisation and peptide degradation. An increase of the relative proportion of sorghum in the grist resulted in decreases in wort filtration, colour, viscosity, attenuation limit, free amino nitrogen, high molecular weight nitrogen, and a corresponding increase in pH (p < 0.01). Overall the addition of malted barley in small proportions to unmalted sorghum mashes together with commercial enzymes was found to improve the potential for brewing a high quality lager beer from unmalted sorghum.     

HEAT SENSITIVITY,OPTIMUM pH AND CHANGES IN ACTIVITY OF SORGHUM PEROXIDASE DURING MALTING AND MASHING

Peroxidase activity was demonstrated in dry and germinating sorghum seeds. The specific activity increased about 14-fold during malting for a 96-hour period. On the average about 41% of peroxidase activity was located in the endosperm, and the remaining 56% in the acrospire and rootlet of sorghum malt. The crude enzyme extract retained 77%, 17.5% and 7.6% of activity after heating at 60°, 70° and 80°C, respectively. More than 50% of the peroxidase activity in the finished malt survived mashing at 65°C. Optimum activity was recorded at pH 5.5 which falls within the observed pH range of sorghum worts. The level of residual peroxidase activity in the wort differed with sorghum species.     

Influence of malted barley and exogenous enzymes on the glucose/maltose balance of worts with sorghum or barley as an adjunct

The sugar profile of wort from laboratory malted barley, malted sorghum, unmalted barley and unmalted sorghum grains mashed with commercial enzyme preparations were studied. Similar levels of glucose to maltose (1:7) were observed in wort of malted barley and malted sorghum. Mashing barley or sorghum grains with commercial enzymes changed the glucose to maltose ratio in both worts, with a greater change in wort from sorghum grains. Although hydrolysis with commercial enzymes released more glucose from maltose in sorghum wort, the same treatment retained more maltose in barley wort. Adding malted barley to sorghum grains mashed with commercial enzymes restored the glucose to maltose ratio in sorghum mash. Fermentation of wort produced from all barley malt (ABM) mash and commercial enzyme/barley malt/sorghum adjunct (CEBMSA) mash of similar wort gravity was also studied. ABM and CEBMSA worts exhibited similar glucose to maltose ratios and similar amino acid spectra. However, ABM released more individual amino acids and five times more proline than wort from commercial enzyme/barley malt/sorghum adjunct. ABM produced 27% more glucose and 7% more maltose than CEBMSA. After fermentation, ABM mash produced 9.45% ABV whilst commercial enzyme/barley malt/sorghum adjunct mash produced 9.06% ABV. Restoration of the glucose/maltose ratio in the CEBMSA mash produced wort with a sugar balance required for high gravity brewing. © 2020 The Institute of Brewing & Distilling     

The effect of germination time on malt quality of six sorghum (Sorghum bicolor) varieties grown at Melkassa,Ethiopia

The effect of germination time (48, 96 and 144 h) on malt quality of six sorghum varieties was investigated to determine the potential of grain sorghum cultivars in the local brewery industry. Six sorghum varieties (Gambella 1107, Macia, Meko, Red‐Swazi, Teshale and 76 T1#23) were evaluated in a randomized complete block design with three replications. The results showed that both germination time and variety had a significant effect (p ≤ 0.05) on sorghum malt quality. The hectolitre weight (kg hL^?1 at 12.5% moisture), germination energy (%), crude protein (%) and flour starch amylose content (%) were: 75.8–82.9, 96–99, 7.0 ? 11.9 and 16.0 ? 23.0, respectively. The sorghum malt diastatic power (DP, °L), free amino nitrogen (FAN, mg L^?1), hot water extract (HWE, %) and malting loss (%) were: 18.96 ? 31.39, 185.67 ? 343.29, 41.85 ? 85.08 and 8.68 ? 27.56, respectively. Malting loss, HWE and FAN increased with germination time. The DP increased as the germination duration increased from 48 to 96 h, but the difference between 96 and 144 h was not significant. Considering the excessive malting loss and marginal increase in HWE beyond 96 h, this study suggests that the optimum malting duration would be around 96 h. Among the varieties tested, Teshale and Gambella 1107 produced the better malt quality. Copyright © 2012 The Institute of Brewing & Distilling     

Pilot Scale Production of a Lager Beer from a Grist Containing 50% Unmalted Sorghum

Pilot scale (1000 L) brews were carried out with a grist comprising of unmalted sorghum (50% of total wet weight of grain) (South African variety) and malted barley (50% of total wet weight of grain) grist using a mashing program with rests at 50°C, 95°C and 60°C. Mashes were supplemented with a high heat stable bacterial α‐amylase, a bacterial neutral protease and a fungal α‐amylase. A control brew containing 100% malted barley was also carried out. Saccharification difficulties were encountered during mashing, and extraction of the grist was lower for the sorghum mashes. The sorghum mashes showed comparable lautering behaviour to that of the control mash. At mashing off the sorghum worts were starch positive. Apparent degree of fermentation of the sorghum gyles were less than the control gyles. Green beer filtration proved unproblematic. The sorghum beers compared quite closely with the control beer with regard to colour, pH and colloidal stability. Foam stability deficiencies were apparent with the sorghum beer. However, the fermentability of the sorghum worts were lower. Hence the sorghum beers were lower in total alcohol. Sensory analysis indicated that no significant differences existed between the sorghum beer and both the control beer and a commercial malted barley beer with regard to aroma, mouth‐feel, after‐taste and clarity. However, the sorghum beer was found to be significantly different to both of the other beers with regard to colour, initial taste and foam stability.     

Effect of varying starch properties and mashing conditions on wort sugar profiles

The aim of this research was to investigate the relationship between starch composition in barley and its malted counterpart alongside malt enzyme activity and determine how these factors contribute to the fermentable sugar profile of wort. Two Australian malting barley varieties, Commander and Gairdner, were sourced from eight growing locations alongside a commercial sample of each. For barley and malt, total starch and gelatinisation temperature were taken, and for malt, α‐ and β‐amylase activities were measured. Samples were mashed using two mashing profiles (infusion and Congress) and the subsequent wort sugar composition and other quality measures (colour, original gravity, soluble nitrogen) were tested. Variety had no significant (<0.05) effect on any barley, malt, enzyme or wort characteristics. However, growing location impacted gelatinisation temperature, colour, malt protein content and original gravity. The gelatinisation temperature in malt samples was higher, by ~0.8°C, than in the equivalent barley sample. Several malt samples, even with protein contents <12.0%, had gelatinisation temperature >65°C. The fermentable sugars measured in the malt prior to mashing showed a higher proportion of maltose than glucose or maltotriose. In addition, there were significant differences in the amount of sugar produced by each mashing method with the high temperature infusion producing a higher amount of sugar and proportionally more maltose. There is scope for further research on the effect of genetics and growing environment on gelatinisation temperature, mash performance and fermentable sugar development. Routinely measuring gelatinisation temperature and providing this information on malt specification sheets could help brewers optimise performance. © 2019 The Institute of Brewing & Distilling     

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 PROPERTIES,COMPOSITION AND FERMENTABILITIES OF WORTS MADE FROM 100% RAW SORGHUM AND COMMERCIAL ENZYMES

A mashing regime was developed using 100% raw sorghum which enabled commercially acceptable hot water extracts to be obtained in 85 minutes with minimal use of a heat stable α-amylase and proteolytic enzymes. This gave worts of HWE 295 1°/kg, with FAN levels of about 40 mg/l and ammonium ion concentration of about 60 mg/l. Higher, but commercially unacceptable, levels of proteolytic enzymes gave worts with FAN from 84.5 to 95 (mg/l). Addition of an amyloglucosidase as the commercial preparation Amylo300L, was required to convert the HWE to fermentable extract. The addition of Amylo300L, increased the DP1, DP2 and DP3 carbohydrate fractions of the worts from 22% to more than 90% of the total, compared to about 80% for a wort made from malted barley without the use of enzymes. Two different proteolytic enzymes gave different extracts and FAN contents presumably reflecting either differences in susceptibilities of the sorghum to the two enzymes or the presence of different additional enzyme activities in the different preparations. The level of ammonium ions in malted barley worts was 86 mg/l and up to 88 mg/l in worts produced from sorghum and enzymes. Enzyme addition produced increased levels of ammonia. The content of Group A (the most readily assimilated) amino acids was proportionally higher in sorghum worts compared to malted barley wort. Worts made from raw sorghum and enzymes, containing as little as 40 mg/l FAN, were fully attenuated. The yeast consumed about 35 mg/l FAN and 45 mg/l ammonium ions. Under identical fermentation conditions, the same yeast, fermenting a malted barley wort of comparable extract consumed 104 mg/l FAN and 37 mg/l ammonium ions.     

Studies on the mashing conditions of teff (Eragrostis tef) malt as a raw material for lactic acid‐fermented gluten‐free beverage

Formation of extracts and fermentable sugars during mashing can be limited by incomplete starch gelatinisation. The aim of this research was to develop mashing programme for 100% teff malt as a potential raw material for gluten‐free lactic acid‐fermented beverage. Isothermal mashing at temperatures ranging between 60 and 84 °C was conducted, and the highest extract (85%) was observed for the wort samples produced at temperatures higher than 76 °C. Sixty‐minute rest at 71 °C resulted in higher fermentable sugars than other tested conversion rest temperatures. Inclusion of lower mashing‐in temperature in the mashing programme also substantially improved the concentrations of free amino nitrogen (128 mg L^?1) and fermentable sugar (58 g L^?1) in the final wort. Therefore, 30‐min rest at 40 °C followed by 60‐min rest at 71 °C and 10‐min rest at 78 °C was found to be a suitable mashing programme for teff malt.     

Influence of High Protein Digestibility Sorghums on Free Amino Nitrogen (FAN) Production during Malting and Mashing

In sorghum brewing, obtaining sufficient Free Amino Nitrogen (FAN) for rapid and complete fermentation remains a problem due to the high proportions of unmalted sorghum used and the poor digestibility of wet‐heat treated sorghum protein. Sorghum mutant lines with high protein digestibility have been developed through breeding. These high protein digestibility sorghums (HPDS) have protein bodies with villi‐like borders that apparently facilitate protease access. This work investigated FAN production from HPDS when malted and mashed, to assess their potential for use in sorghum brewing to improve wort FAN levels. When malted, HPDS contained substantially higher levels of FAN than normal protein digestibility sorghums (NPDS), 32 mg/100 g malt more. However, when the HPDS were mashed either as malt, or as grain or malt plus exogenous proteases, FAN production during mashing was not substantially higher than with NPDS subjected to the same treatments, only 6, 6–18 and 9–13 mg/100 g grain or malt, respectively. This is probably due to wet‐heat induced cross‐linking of the kafirin proteins reducing their susceptibility to proteolysis. Notwithstanding this, HPDS could be very useful for improving FAN levels in sorghum brewing if they are malted.     

Optimization of the sorghum malting process for pito production in Ghana

Pito is an alcoholic beverage obtained through a yeast (Saccharomyces cerevisiae) fermentation of wort extracted from sorghum (Sorghum bicolor L. Moench) malt. The malting conditions of sorghum are thought to influence the quality characteristics of the malt, and subsequently the quality of the pito obtained from it. Studies were carried out on a local sorghum cultivar grown in Ghana – chireh, to optimize the conditions for malting conditions for pito production in Ghana. A 3³ full factorial experimental design was replicated with steeping times of 12, 16 and 22 h, germination times of 3, 4 and 5 days, and malt drying temperatures of 30, 40 and 50 °C as factors. Diastatic power, extract yield, attenuation limit and free amino nitrogen were determined. Germination duration significantly affected diastatic power and free amino nitrogen (p < 0.001). Extract yield was also significantly influenced by germination duration (p = 0.001). The germination time, steeping time and drying temperature had no significant effect on the attenuation limit. The optimal conditions for malting this specific cultivar grown in Ghana to obtain critical malt quality indices are 12.0–12.5 h steeping, 5 days of germination at 30 °C and drying at 40 °C. Free amino nitrogen levels in all treatments were higher than the minimum requirement for good yeast nutrition and fermentation. Copyright © 2015 The Institute of Brewing & Distilling     

Changes in Germination and Malting Quality During Storage of Barley

To increase brewing yield and efficiency, malts with high extract values, high enzymic activities and good modification are essential. To produce malt that meets these requirements, the barley employed must have minimal post‐harvest dormancy and be able to germinate vigorously. The aims of this study were to determine the extent to which some Australian barley varieties changed during post‐harvest storage, how these changes influenced germination characteristics, enzyme production and malt quality, and, of the germination tests examined, which gave the best indication of a barley's malting potential. Four commercially grown barley samples were obtained, one from Tasmania and three from Victoria. Each sample was stored at room temperature for one year. At monthly intervals, samples were taken and placed at ?18°C. The germinative energy (GE) and germinative index (GI) of these samples were measured. Samples were also micro‐malted and the quality of the malt was assessed using standard EBC methodology. Storage at room temperature positively influenced the germination characteristics of all samples, with concomitant improvements in hydrolytic enzyme production during malting and in a number of malt quality parameters. It was found that, of the germination tests examined, the GI consistently correlated with enzyme activities during malting and with various malt quality parameters thus indicating that the GI is a good indicator of malting potential.     

Quality Assessment of Different Sorghum Varieties for Their Brewing Potential

Four sorghum varieties (SK 5912, KSV 4, KSV 8, ICSV 400) were malted and extracted under similar conditions to assess their quality for brewing. The results showed that, in general, the sorghum varieties had high malting loss which was attributed to the high germination temperature used. The sorghum varieties also developed low levels of amylolytic activity (α‐amylase and β‐amylase), and with similar ratios. When the sorghum malts were mashed at different temperatures with the aid of commercial enzyme preparations, it was observed that mashing temperatures were more important in sugar release than additions of commercial enzymes. This was because at the lower mashing temperature, sorghum starch was not adequately gelatinised. However, when commercial enzyme preparations were added, low levels of enzymes were very effective in reducing wort viscosity and producing free amino nitrogen (FAN). Although, both commercial enzyme preparation and mashing temperature influenced sugar production, the malts produced glucose and maltose at similar ratios. Therefore good quality malts can be produced from sorghum, however mashing will employ commercial enzymes and mashing regimes are not yet optimised.     

STUDIES ON THE EFFECT OF GERMINATION TIME AND TEMPERATURE ON MALTING OF RICE

The effect of germination conditions on rice malt quality was studied by germinating rice for different periods of time up to 7 days over a range of temperatures (22° to 32°C). Germination time and temperature had a significant effect on rice malt diastatic power, cold and hot water extracts, total reducing sugars, and free alpha amino nitrogen. In general, diastatic power, cold water extract, hot water extract, total reducing sugars and free alpha amino nitrogen, all increased with germination time and temperature. Germination temperatures of 28°C and 30°C were good for the development of diastatic power, cold water extract, hot water extract, total reducing sugars and free alpha amino nitrogen. However germinating at 30°C gave maximum result. The mashing trials with rice malted for 6 days showed a slow and incomplete saccharification, slow filtration rate, and high total available extract in spent grains.