Sorghum β‐Amylase Production: Relationship With Grain Cultivar,Steep Regime,Steep Liquor Composition and Kilning Temperature

Steep regime, nature of alkaline liquor, and kilning conditions were studied for their effects on sorghum malt β‐amylase development in four Nigerian sorghum cultivars. Malt β‐amylase activity was markedly (p < .001) influenced by all the four factors as well as their various interactions. Overall, malts from KSV 8 variety were the most β‐amylolytic followed in sequence by those from Local Red (LR), SK 5912, and Local white (LW) respectively. The presence or absence of air rests in steep regimes was a significant (p < .001) determinant of sorghum β‐amylase response to final warm steeping, steep liquor and kilning condition. The nature of the alkaline steep liquor was also a major determinant of the pattern of malt β‐amylase response to the kilning condition. Steeping in Ca(OH)₂ enhanced malt β‐amylase activity at the higher temperature of kilning, while KOH produced the opposite effect. Ca(OH)₂ enhancement of β‐amylase development, at a kilning temperature of 50°C, was variety‐dependent suggesting that different sorghum cultivars may employ different biosynthesis models for β‐amylase synthesis. The regime‐dependence of β‐amylase response to kiln temperature suggests that this was an important modulator of sorghum germination physiology.     

A Simulation Model for the Control of beta‐Glucanase Activity and beta‐Glucan Degradation During Germination in Malting

A dynamic model was developed that describes the formation of β‐glucanase and the degradation of β‐glucans at different temperatures and grain moistures during the germination of malting barley. The process was analysed by simulations and by solving an optimal control problem for maximising the β‐glucanase activity. The results demonstrate the effects of controlling dynamically the germination process and improve the understanding of cytolysis in germination.     

A review of malting and malt processing for whisky distillation

This paper encompasses a re‐evaluation of published literature and data regarding wort attenuation in malt distilleries raising questions and discussing how the conventional wisdom has changed over time and what questions still need to be answered. Current knowledge is summarized in the following four points: (a) Under normal malting conditions, starch granules are partially degraded by a combination of α‐amylase and α‐glucosidase. This complex can open up the granule at specific sites on the surface and create characteristic ‘pin‐hole’ lesions, which may be widened by secondary hydrolysis by α‐ and β‐amylase, limit dextrinase and α‐glucosidase (maltase). (b) All of these diastatic enzymes can survive mild kilning, probably by forming heat stable complexes on and within the starch granules and can continue a complete degradation of starch when mashed at ambient temperatures with glucose as the end product. (c) At normal mashing temperatures, starch granules gelatinize and dissolve with a concomitant rapid degradation to glucose, maltose, maltotriose and dextrins ranging from degree of polymerization (DP) 4 to > DP20. If there is immediate wort boiling after run‐off, this is the final composition of starch derived carbohydrates according to the conventional paradigm. (d) All malt worts also contain a small amount of panose, isopanose as well as glucosyl maltodextrins, based on a core of 6₂‐α‐glucosyl maltose (panose) or 6‐α‐maltosyl glucose (isopanose), which are remnants of the α‐amylase/glucosidase degradation of granular starch. These dextrins are resistant to the action of debranching enzymes and their concentration may vary between 4 and 8% of the malt extract, depending on the degree of modification of the host starch granules. They may be created at the active sites of this enzyme complex when the granule is gelatinized. In a conventional mash of unboiled distilling wort, the spectrum of wort dextrins produced from gelatinized starch is reduced to true ‘limit’ dextrins of DP4–8 by continued α‐amylolysis during early fermentation. These dextrins will contain side chains of either maltose or maltotriose residues surrounding the α‐1,6‐glucosidic linkage and can be debranched by limit dextrinase during late fermentation, leaving only the above glucosyl maltodextrins dextrins in the spent wash. Copyright © 2016 The Institute of Brewing & Distilling     

Effect of High Hydrostatic Pressure‐Temperature Combinations on the Activity of β‐Glucanase from Barley Malt

β‐Glucanase from barley malt is known to be thermolabile but important in the mashing process. Therefore, the potential of increasing the thermostability of β‐glucanase in ACES buffer (0.1 M, pH 5.6) by high hydrostatic pressure has been investtigated. Inactivation of the enzyme as well as changes of the conversion rate in response to combined pressure‐temperature treatments in the range of 0.1–900 MPa and 30–75°C were assessed by analyzing the kinetic rate constants. A significant stabilization of β‐glucanase against temperature‐induced inactivation was detected at 400 MPa. With increasing pressure up to 600 MPa the catalytic activity of β‐glucanase was progressively decelerated. However, for the overall depolymerization reaction of β‐glucans in ACES buffer (0.1 M, pH 5.6) a maximum was identified at 215 MPa and 55°C yielding approximately 2/3 higher degradation of β‐glucan after 20 min as compared to the maximum at ambient pressure (45°C).     

Effects of Wheat Protein Content on Endosperm Composites and Malt Quality

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

Properties of Three Sorghum Cultivars Used for the Production of Bili‐Bili Beverage in Northern Cameroon

The malting characteristics of sorghum malts produced locally in Cameroon for Bili‐Bili brewing were compared with those of malts produced in a laboratory. The analytical values of both malts were similar but the brewing potential of the laboratory malts were marginally better than those of the locally produced malts. Of the three cultivars examined, Madjeru had the lowest levels of β‐amylase, maltose levels and fermentability. The worts of the Madjeru filtered the slowest of the three malts. During malting β‐glucanase developed rapidly and development was temperature‐dependent.     

Effects of Pulses of Higher Temperature on the Development of Enzyme Activity During Malting

The increase of temperature at the beginning, in the middle and at the end of malting has been evaluated in terms of quality parameters (malting losses, index of acrospire development, friability, HWE, viscosity, SNR) and enzyme (β‐glucanase and α‐amylase) development, in a good quality malting barley (Otis) and a higher protein‐higher β‐glucan content barley used for feed (Extra). A shift from 15 to 20°C at the beginning of malting was shown to increase acrospire development, friability, HWE and SNR and to reduce viscosity, without significantly affecting malting losses. This effect was related to higher β‐glucanase and α‐amylase activities within each variety. However, the same enzyme activities were not directly related to a better malting quality when the two genotypes were compared. This confirms previous indications that diversity in malting performance between genotypes cannot simply be traced back to differences in enzyme activities; but, indeed, it suggests that, for a defined barley lot, changes in the levels of enzyme activities following different malting procedures may have a direct effect on malt quality.     

Malting of Sorghum: Further Studies on Factors influencing α‐Amylase Activity

The effects of steep regime, nature of alkaline steeping agent, and kilning condition on α‐amylase development were studied for four Nigerian sorghum cultivars. Malt α‐amylase activity was highly significantly (p<.001) influenced by all the four factors as well as their various assortments of interaction. Generally malts from the Local Red (LR) variety produced the highest a‐amylase values, followed by those of SK 5912, Local White and KSV 8 in the above sequence. The presence/absence of air‐rest processes in steep regimes was a significant factor (p<.001) influencing malt α‐amylase response to final warm steeping as well as to the other factors under study. Similarly, the nature of the steeping agent was a very significant determinant of malt α‐amylase response to kilning condition and regime of steeping. Of significant interest was the observation that Ca (OH)₂ steeps enhanced malt α‐amylase activity at the higher temperature of kilning. The significantly lower α‐amylase values given under similar conditions by the other alkaline liquors suggest a possible increase in malt thermostability due to steeping in Ca (OH)₂. Additionally, the fact that the extent of enhancement of malt α‐amylase activity by Ca (OH)₂, at 50°C Kiln temperature, was regime‐dependent, suggests that the latter was an important modulator of sorghum germination physiology.     

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.     

PURIFICATION AND PARTIAL CHARACTERIZATION OF AN ENDO-β1,3-GLUCANASE FROM GREEN MALT

An endo-β1,3-glucanase from a green malt extract was purified by DEAE- and CM-cellulose ion exchange chromatography followed by molecular sieve chromatography on BioGel P-100. A final enzyme preparation had two protein components on disc electrophoresis, one of which was in-active. The enzyme had a pH optimum of 5·0 for activity on laminarin and 5·8 on carboxymethyl pachyman. The activity was stable up to 60°C and was stimulated by NaCl. The isoelectric point of the enzyme was 9·8.     

Malt Performance is More Related to Inhomogeneity of Protein and β‐Glucan Breakdown than to Standard Malt Analyses

Nitrogen analyses of the grains of samples of commercial malts indicate that β‐glucan breakdown and the uniformity of malt modification are influenced by uniformity of distribution of grain protein. It is proposed that for normal malting barley, variations in malt modification are related to the different percentages of grains which contain high levels and different types of proteins which resist enzymic modification during malting. This kind of inhomogeneity of malt modification can cause brewhouse problems but cannot be detected with precision by standard malt analyses.     

AN INTER-LABORATORY STUDY OF STARCH DEGRADING ENZYME ASSAYS

The reliability of specific colorimetric assays for the main starch degrading enzymes, α-amylase, β-amylase and limit dextrinase was investigated by conducting an inter-laboratory collaborative study. Three malt samples were assayed in duplicate in 14 laboratories. The repeatability (r) within laboratories was good for α-amylase and β-amylase assays. Between-laboratory reproducibility (R) was acceptable (CV_R 9.2–14%). Errors for limit dextrinase were higher. Repeatability was good at high enzyme concentrations (CV_r = 2%) but increased at low enzyme concentrations. Reproducibility was relatively poor (CV_R-27–33%).     

Enzyme activities,dietary fibre components and rheological properties of wholemeal flours from rye cultivars grown in Finland

Total and soluble dietary fibre, total and soluble pentosan and β‐glucan contents, activities of α‐amylase (EC 3.2.1.1) and endo‐β‐xylanase (EC 3.2.1.8) and viscous properties of aqueous suspensions of wholemeal flours during heating were determined in nine winter rye cultivars (Secale cereale L) grown in Finland in 1998–2001. There was marked annual and varietal differences in grain quality. In the rainy summer 1998 the yield was low, grains were small and dietary fibre content of the grains was high. Xylanase activity of the grains was high, which corresponded to the high content of soluble pentosans. In the dry summer of 1999, the pentosan content of the grains was low and β‐glucan content high. The effect of weather conditions and cultivar were also apparent in the differences in falling numbers, amylogram and swelling curve results. The two hybrid rye cultivars Esprit and Picasso had consistently highest falling numbers and amylogram peak viscosities. The activities of α‐amylase and xylanase had a moderate positive correlation with total pentosan content and the content of soluble pentosan. Xylanase activity had better correlation with the viscous properties of flour–water suspensions than α‐amylase. Surprisingly, α‐amylase activity had only a moderate negative correlation with falling number. Copyright © 2005 Society of Chemical Industry     

OPTIMIZED McCLEARY METHOD FOR MEASUREMENT OF TOTAL β-AMYLASE IN BARLEY AND ITS APPLICABILITY

The McCleary method for determination of β-amylase in malt has been modified in order to allow determination of total β-amylase in barley as well as malt. A ruggedness test, performed on the modified method, demonstrated that the method is quite robust and highly reproducible. When the variables α-amylase, β-amylase and diastatic power were measured in 90 malt samples, only β-amylase was significantly correlated to diastatic power (r² = 0.85 and p < 0.0001). The same high correlation was found between total β-amylase in 20 barley samples and diastatic power in the corresponding malts. The validity of this relationship was tested by predicting diastatic power in malt from total β-amylase in barley. Predicted values correlated highly to measured values (r² = 0.95). In breeding material a positive relationship was found between total β-amylase in barley and protein content. This relationship must be considered when evaluating new barley lines.     

Molecular Weight Affected Antioxidant,Hypoglycemic and Hypotensive Activities of Cold Water Extract from Pleurotus citrinopileatus

Cold water extract of P. citrinopileatus (CWEPC) was fractioned into 4 fractions, PC‐I (<1 kDa), PC‐II (1‐3.5 kDa), PC‐III (3.5‐10 kDa), and PC‐IV (>10 kDa), by ultrafiltration. The antioxidant activities, the inhibition of pancreatic α‐amylase, intestinal α‐glucosidase, and hypertension‐linked angiotensin converting enzyme (ACE), as well as the contents of polysaccharides, protein, and phenolic compounds of 4 fractions were determined. The results showed that lower MW fractions exerted a higher antioxidant activity, which was correlated to phenolic contents. The high molecular fraction (PC‐IV) exhibited significantly higher inhibitory activity on α‐amylase, α‐glucosidase, and ACE compared to CWEPC and the other 3 lower MW fractions (<10 kDa), which was more related to protein contents. The inhibition capability of CWEPC and PC‐IV on α‐amylase activity was 1/13.4 to 1/2.7 relative to that of acarbose, respectively. Kinetic data revealed that PC‐IV fraction followed a noncompetitive inhibition pattern on α‐glucosidase activity. The study demonstrated that various MW fractions and types of components contribute to different biological functions of P. citrinopileatus and it is protein constituents but not peptides responsible for the hypoglycemic potential of CWEPC.     

Enzymatic Properties of β‐1,3‐Glucanase from Streptomyces sp Mo.

Streptomyces sp Mo endo‐β‐1,3‐glucanase was found to have hydrolyzing activity toward curdlan and released laminarioligosaccharides selectively. The molecular weight was estimated to be 36000 Da and its N‐terminal amino acid sequence was VTPPDISVTN. The optimal pH was 6 and the enzyme was found to be stable from pH 5 to 8. The optimal temperature was 60 °C and the activity was stable below 50 °C. The enzyme hydrolyzed selectively curdlan containing only β‐1,3 linkages. The enzyme had 89% relative activity toward Laminaria digitata laminarin, which contains a small amount of β‐1,6 linkages compared with curdlan, while Eisenia bicyclis laminarin with a higher amount of β‐1,6‐linkages, was not hydrolyzed. Mo enzyme adsorbed completely on curdlan powder. The enzymatic hydrolysis of curdlan powder resulted in the accumulation of laminaribiose (yield 81.7%). Trisaccharide was inevitably released from the hydrolysis of laminarioligosaccharides with 5 to 7 degrees of polymerization (DP). Although the enzyme cleaved off disaccharide (DP 2) from tetrasaccharide (DP 4), the reaction rate was lower than those of DP 5 to 7. The results indicated that the active site of Mo endo‐β‐1,3‐glucanase can efficiently recognize glucosyl residue chain of greater than DP 5 and hydrolyzes the β‐1,3 linkage between the 3rd and 4th glucosyl residue.     

Malting Performance of Normal Huskless and Acid‐Dehusked Barley Samples

Sulphuric acid dehusked barley had a higher germinative energy and lower microbial infection than normal huskless (naked) barley, suggesting that the pericarp layer harboured microbial infection which may have limited the germination rate. Dehusking the normal huskless barley using sulphuric acid resulted in lower microbial infection, and increased germinative energy. The normal huskless barley sample had a higher β‐glucan content than the acid‐dehusked barley and had slower β‐glucan breakdown during malting. This resulted in the release of seven times more β‐glucan during mashing, and the production of wort of higher viscosity. The normal huskless barley sample had a higher total nitrogen content than the acid‐dehusked barley but both samples produced similar levels of amylolytic (α‐ and β‐amylase) activity over the same malting period. No direct correlation was found between barley total nitrogen level and the amylolytic activity of the malt. When barley loses its husk at harvest, the embryo is exposed and may be damaged. This may result in uneven germination which can reduce malting performance and hence malt quality.     

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.