RECOMMENDED METHODS OF ANALYSIS OF BARLEY,MALT AND ADJUNCTS ALTERNATIVE TITRATION PROCEDURE FOR THE DETERMINATION OF THE DIASTATIC ACTIVITY OF MALT

The Sub-Committee was unable to recommend the determination of wort fermentability as an alternative to the determination of diastatic activity in the evaluation of malts. Potassium ferricyanide solution is recommended as an alternative to Fehling's solution in the determination of the diastatic power of malt because the titration is simpler and the reproducibility of the determination is less dependent on the volume of cold water extract taken for diastasis.     

FACTORS CONTROLLING THE FERMENTABLE EXTRACT IN DISTILLERS MALT

The relationships between the alcohol yield obtainable from different malts and some readily measurable parameters have been investigated. Gel filtration studies clearly show that melanoidin-type condensation products, which were previously suggested as a cause of low fermentability, are not produced in significant amounts in highly modified malts. These studies also show important details relating to the hydrolysis of dextrins in distillers' worts and washes. Measurement of the limit dextrinase activity of a range of distillers' malts has indicated that this enzyme is not a limiting factor in determining wort fermentability and its concentration does not correlate with the overall spirit yield. There may be a relationship between the whole amylase complex, as expressed by the diastatic power and wort fermentability. From a study of the reducing sugar contents of worts, it was confirmed that the major factor controlling fermentable extract in malt is the relative amounts of starch-derived sugars in the wort. It now appears possible to determine the fermentability of distilling wort by the standard extract and the reducing power of the wort obtained by the standard I.O.B. extraction.     

THE DETERMINATION OF FERMENTABLE EXTRACT IN BREWING WORTS

A method for determining the real fermentability of a standard Institute of Brewing wort is proposed for use in the Brewing Industry. The method is based on a system currently operating in the Distilling Industry using standard Distillers Co. Ltd (D.C.L.) ‘M’ type yeast. To provide a safeguard against variability in the performance of the yeast used, it is recommended that a standard malt (e.g. Institute of Brewing check malt A) of known fermentable extract should be used to check the condition of the yeast.     

THE EFFECTS OF MASHING TEMPERATURE AND MASH THICKNESS ON WORT CARBOHYDRATE COMPOSITION

Temperature and mash thickness are shown to affect both mash performance and enzyme activity. Alpha amylase was found to be considerably more resistant to heat inactivation than was beta amylase. This difference was reflected by changes in wort fermentability that were manifest at temperatures below those which affected levels of extract. Increasing the mashing temperature from 65°C to 80°C had only a slight effect on extract but reduced wort fermentability from over 70% to less than 30%. At 85°C and over, when temperature had a significant effect on alpha amylase, as well as on beta-amylase, extract was lost and starch was present in the wort. Diluting the mash with liquor had a similar effect to that of increasing temperature on both the amylolytic enzymes and on the mash performance. Thin mashes contained more starch and fewer fermentable sugars than did thick mashes at the same temperature. These changes can be related to the stability of the amylolytic enzymes.     

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.     

Importance of Endosperm Modification for Malt Wort Fermentability1

Malt wort fermentability is dependent on an adequate supply of the essential nutrients required by yeast. The barley and malt factors affecting this supply of nutrients are not well understood. This study used two doubled haploid populations (Arapiles × Franklin, TR251 × HB345), the latter with a hulless barley parent, to investigate effects of barley and malt quality on fermentability. Populations were grown and malted at different locations resulting in a broad range in malt quality with significant differences in extract, modification and enzyme levels. Fermentability, as indicated by apparent attenuation limit and ethanol levels, also showed significant differences among samples. Modification was the most important factor for good fermentability. There appeared to be several different mechanisms by which modification affected fermentability. High viscosities, slow starch release during mashing, increased glucose supply from better β‐glucan breakdown and increased free amino nitrogen levels all affected fermentability. Effects of starch‐degrading enzymes on fermentability became more significant in better modified malts with α‐amylase showing stronger effects than diastatic power. The poorer fermentability of hulless barley malt was predominately due to low levels of α‐amylase, although, free amino nitrogen also appeared to be an important factor.     

Enhanced NIR Calibration for Wort Fermentability Using Orthogonal Signal Correction

An enhanced method for the calibration of Near Infra Red (NIR) reflectance spectra to wort fermentability is proposed using a signal pre‐processing algorithm called orthogonal signal correction (OSC). Pre‐processing NIR spectra prior to partial least squares Project to Latent Structures (PLS) regression modelling is becoming commonplace in multivariate calibration. A set of twenty wort samples subjected to a replicated 2² factorial design with a centre point and nine production samples were used to construct multivariate prediction models. The experimental design factors were the mash tun saccharification temperature and time used to purposely provide a sample set with significant leverage in the fermentability responses. Calibration PLS models for both wort apparent degree of fermentation (ADF) and final attenuation apparent extract (Final AE) values with and without OSC corrected spectra were compared demonstrating significant improvements in prediction capability with the prior (Q² = 0.90 versus Q² = 0.28). The OSC algorithm removed almost 60% of the variance in the NIR spectra, which was independent or orthogonal to the fermentability measures. By cleaning up the spectra, the standard errors of prediction (SEP) for ADF and Final AE were improved by 50 and 90%, respectively, illustrating not only the enhancement in calibration but also the aptness for process control applications. Various model validation tests, including an external validation example and random response permutation, verify the validity of the models using OSC. Furthermore, interpretation of the important wavelengths related to wort fermentability is provided and demonstrates that some key wavelengths are related to both carbohydrate overtones as well as nitrogen functional groups. The application of OSC prior to developing calibration models with NIR demonstrates promising results for brewers interested in real time control of wort fermentability.     

Use of Response Surface Methodology to Investigate the Effectiveness of Commercial Enzymes on Buckwheat Malt for Brewing Purposes

The influence of two factors, total concentration and fraction of three pairs of commercial enzymes, which showed statistical significance (Biocellulase W with Hitempase 2XL, Biocellulase W with Amylo 300 and Amylo 300 with Hitempase 2XL), were studied for their overall effect on buckwheat wort quality using response surface methodology (RSM). This study revealed that the addition of increasing levels of Hitempase 2XL to the buckwheat mash increased colour, extract levels, wort filtration, fermentability and total fermentable extract (TFE), along with decreasing viscosity values. Results also determined a high level of fermentability when an enzyme combination of 30% Biocellulase and 70% Hitempase was added to the mash. The addition of increasing levels of Amylo 300 to buckwheat mashes resulted in increases in fermentability and total fermentable extract (TFE), along with increases in total soluble nitrogen (TSN), free amino nitrogen (FAN) and Kolbach index (KI). With regard to the proposed optimal regime, although no synergistic effect was found when the three enzymes were used together, the optimum conditions for the production of buckwheat wort with lowest viscosity, highest extract and optimal fermentability were achieved using a joint model. Overall, the findings of this study demonstrate the feasibility of producing wort suitable for the brewing of gluten‐free beer from 100% malted buckwheat with careful optimisation of enzyme types and dosage levels.     

Validation and application of osmolyte concentration as an indicator to evaluate fermentability of wort and malt

The objective was to develop a new simple and quick approach to predict fermentability, based on osmolyte concentration (OC). Eight malts were assayed for diastatic power, starch‐degrading enzymes [α ‐amylase, β ‐amylase and limit dextrinase (LD)] and malt OC (MOC). All malts were mashed to determine wort OC (WOC), real degree of fermentation (RDF) and sugar contents in a small‐scale mashing protocol. The results showed that MOC was correlated with malt α ‐amylase, LD, the resultant WOC, RDF and fermentable sugar (r  = 0.813, 0.762, 0.795, 0.867, 0.744, respectively), suggesting that MOC was discriminating in predicting levels of malt amylolytic enzymes, wort sugar and RDF without the mashing and fermentation process. Moreover, WOC showed stronger correlations with malt α ‐amylase, LD, RDF and fermentable sugars (r  = 0.796, 0.841, 0.884, 0.982, respectively), suggesting that WOC can be used to quickly predict wort sugar contents and RDF without a fermentation step. Furthermore, the effects of mashing temperature and duration on WOC, RDF and sugar contents are discussed. Adjusting mash temperature to 65°C or extending the mash duration dramatically increased RDF and WOC, whereas malt extract was relatively stable. Similarly, WOC showed significant correlations with RDF and fermentable sugars (r  = 0.912 and 0.942, respectively), suggesting that WOC provides a simple and reliable tool to assist brewers to optimize mash parameters towards the production of ideal wort fermentability. In conclusion, the ability of OC to predict malt fermentability and sugar content allows brewers to keep better control of fermentability in the face of variation of malt quality, and to quickly adjust mashing conditions for the consistency of wort fermentability. Copyright © 2017 The Institute of Brewing & Distilling     

Changes in malt quality during production in two commercial malt houses

This investigation presents a holistic and comprehensive assessment of the stepwise changes in barley quality during the malting process for multiple batches of two Australian malting varieties (Buloke and Gairdner), in two modern, commercial scale pneumatic malthouses. The study sought to analyse and compare malting plant and variety with respect to basic changes in malt quality for protein (total protein and free amino nitrogen), fermentability (apparent attenuation limit and diastatic power), extract yield, along with filtration indicators (lautering efficiency, viscosity and β-glucan). Overall, comparing the two malt plants, it was observed that although malt batches and varieties followed different malting pathways, the finished and kilned malt  was of satisfactory quality in terms of FAN, viscosity, friability, fermentability and extract. © 2020 The Institute of Brewing & Distilling     

DETERMINATION OF FERMENTABILITY OF UNBOILED MALT WORTS A METHOD FOR USE IN THE DISTILLING INDUSTRY

A method for determining the real fermentability of unboiled Institute of Brewing Hot Water Extract (wort) has been assessed by the Analysis Committee and is recommended for use in the Distilling Industry.     

EFFECT OF MALTING TEMPERATURE AND TIME ON ENZYME DEVELOPMENT AND SORGHUM WORT PROPERTIES

The effect of malting temperature and time on enzyme development and wort properties of an improved Nigerian sorghum cultivar (Ex-Kwara) were investigated. Malting was carried out at two temperature regimes, 20°C and 25°C for eight days. Parameters evaluated included α- and β;-amylase development, hot water extract (HWE), soluble extract, fermentability, fermentable extract, viscosity, filtration rate, reducing sugars, α;-amino nitrogen and total soluble nitrogen (TSN). For virtually all the parameters studied, germination at 25°C produced higher values on the 4th day after which temperature appeared to have little influence. α;-Amylase development continued throughout the germination period while β;-amylase peaked on the 6th day. Optimal values of total soluble nitrogen (TSN) were recorded at both 25°C and 20°C at the 6th and 8th day of germination respectively .     

Comparison of the impact on the performance of small‐scale mashing with different proportions of unmalted barley,Ondea Pro®, malt and rice

The impact of using different combinations of unmalted barley, Ondea Pro® and barley malt in conjunction with a 35% rice adjunct on mashing performance was examined in a series of small scale mashing trials. The objective was to identify the potential optimal levels and boundaries for the mashing combinations of barley, Ondea Pro®, malt and 35% rice (BOMR) that might apply in commercial brewing. Barley and malt samples used for the trials were selected from a range of Australian commercial barley and malt samples following evaluation by small‐scale mashing. This investigation builds on previous studies in order to adapt the technology to brewing styles common in Asia, where the use of high levels of rice adjunct is common. Mashing with the rice adjunct, combined with differing proportions of barley, Ondea Pro® and malt, resulted in higher extract levels than were observed for reference mashing, using either 100% malt reference or 100% barley reference and Ondea Pro® enzymes. Synergistic mashing effects between barley, Ondea Pro® and malt were observed for mash quality and efficiency parameters, particularly wort fermentability. The optimum levels of barley in the grist (with the relative level of Ondea Pro®) were assessed to be in the range 45–55% when paired with 10–20% malt and 35% rice. When the proportion of malt was reduced below 10% of the grist, substantial reductions in wort quality were observed for wort quality parameters including extract, lautering, fermentability, free amino nitrogen and haze. Extension of this new approach to brewing with rice adjuncts will benefit from further research into barley varietal selection in order to better meet brewer's quality requirements for the finished beer. Copyright © 2016 The Institute of Brewing & Distilling     

Effect of different analysis conditions on Rapid Visco Analyser malt viscograms in relation to malt of varying fermentability

Malt fermentability is a difficult and time consuming trait to measure. The Rapid Visco Analyser (RVA) was assessed as an alternative rapid method to indicate potential fermentability. This study evaluated changes in rheological profiles under different operational conditions and compared these changes with reference to malt fermentability from a limited number of samples. Viscosity measurements of samples were also made using different RVA (models 3D+ and 4) analysis conditions including a brewhouse time–temperature profile, heating/cooling rate, particle size and enzyme activity. Rheological measurements using the RVA‐3D + gave similar results compared with the RVA‐4, indicating adequate sensitivity of the RVA 3D+ for discriminatory purposes. Use of a time–temperature profile of a commercial brewery mashing process was unsuitable. When malt enzymes were inactivated with silver nitrate, differences in viscosities were observed. However, this eliminated the ability to discriminate on fermentability. Increasing or decreasing the heating rate influenced the time available for enzyme action, which affected the degree of discrimination. This also provided some insight into physical and biochemical processes affected by differences in particle size. RVA has the potential to be used as a tool to discriminate between poor and good fermentability barley malts. RVA conditions when using the ‘Kilned Malt’ method with an appropriate mashing malt–water ratio provided a fast and reliable indication of malt performance prior to conducting lengthy fermentability tests. Copyright © 2014 The Institute of Brewing & Distilling     

EFFECTIVE TRANSPORTATION OF BREWER'S YEAST SLURRY

A yeast shipment method suitable (depending on vessel size) for 1 to 20 litres of slurried yeast is described. The method was investigated primarily assuming transit (storage) temperature at approximately 21°C. Under these conditions, the contained yeast was stable over 116 h at temperatures well under 16°C, and maintained its viability, fermentability and vigour to reinitiate new fermentations. Under more vigorous climatic conditions, shipment time would have to be reduced, but adequate protection time was afforded for all but the most extreme sub-zero situations.     

A New Approach to Limit Dextrinase and its Role in Mashing*

Limit dextrinase (EC 3.1.2.41) is a debranching enzyme catalyzing the hydrolysis of α-1,6-glucosidic linkages in starch. The role of this debranching enzyme in beer brewing has been questioned due to its assumed heat lability. In the present work the effectiveness of limit dextrinase was studied under conditions mimicking brewery practice rather than in a buffer solution. It was demonstrated that typical conversion temperatures of 63–65 °C and a mash pH of 5.4–5.5 favour the action of malt limit dextrinase. The temperature optimum for the limit dextrinase of a malt extract was 60–62.5 °C, as opposed to 50 °C for purified limit dextrinase. Lowering the mash pH from 5.8 to 5.4 increased wort fermentability due to increased limit dextrinase activity. Wort fermentability was more strongly correlated to the free limit dextrinase activity of malt than to the α- and β-amylase activities.     

Refining the Prediction of Potential Malt Fermentability by Including an Assessment of Limit Dextrinase Thermostability and Additional Measures of Malt Modification,Using Two Different Methods for Multivariate Model Development

Prediction of malt fermentability (apparent attenuation limit — AAL) by measurement of the diastatic power enzymes (DPE), α‐amylase, total limit dextrinase, total β‐amylase, β‐amylase thermostability, and the Kolbach index (KI or free amino nitrogen — FAN) is superior to the conventional use of diastatic power (DP) alone. The thermostability of β‐amylase is known to be an important factor in determining fermentability, thus the thermostability of the other relatively thermolabile enzyme, limit dextrinase, was investigated to determine if it was also useful in predicting fermentability. To facilitate this aim, methods were developed for a rapid and cost efficient assay of both β‐amylase and limit dextrinase thermostability. Internationally important Australian and international malting varieties were compared for their total limit dextrinase and β‐amylase activity and thermostability. Interestingly, the level of limit dextrinase thermostability was observed to be inversely correlated with total limit dextrinase activity. The prediction of malt fermentability was achieved by both forward step‐wise multi‐linear regression (MLR) and the partial least squares (PLS) multivariate model development methods. Both methods produced similar identifications of the parameters predicting wort fermentability at similar levels of predictive power. Both models were substantially better at predicting fermentability than the traditional use of DP on its own. The emphasis of this study was on the identification of predictive factors that can be consistently used in models to predict fermentability, because the model parameter estimates will subtly vary depending on mashing conditions, yeast strain/fermentation conditions and malt source. The application of these multivariate model development methods (PLS and MLR) enabled the identification of further potential fermentability predicting factors. The analyses divided the predictive parameters into those defined by DP enzymes and those associated with modification (KI, FAN, fine/coarse difference, wort β‐glucan and friability). Surprisingly, limit dextrinase thermostability was not a substantial predictor of fermentability, presumably due to its negative correlation with total limit dextrinase activity. The application of these insights in the malting and brewing industries is expected to result in substantial improvements in brewing consistency and enable more specific quality targets for barley breeder's progeny selection cut‐off limits to be more precisely defined.     

THE USE OF FORMALDEHYDE IN RESTEEPING

Small-scale malting trials showed that resteeping in a solution of formaldehyde was more uniformly effective than resteeping in water in reducing rootlet yield and the overall malting loss. The hot water extract, the fermentability of the wort, and the formol nitrogen were the same in worts from malts prepared by resteeping in water or formaldehyde solutions. However, cold water extract was reduced, as was the total soluble nitrogen, by the presence of formaldehyde. The anthocyanogen level in the wort was reduced by about 73% when formaldehyde was used in the resteep liquor. The residue of formaldehyde in the unboiled wort was about I p.p.m. It is considered that, in view of the improved resteeping performance and the expected increase in beer stability resulting from the reduction in anthocyanogens and wort nitrogen, the use of formaldehyde in resteeping is worth evaluating on a larger scale. In addition the use of formaldehyde in the resteep liquor prevented fouling by micro-organisms and resulted in a cleaner malt.     

THE APPLICATION OF TRANSMISSION NEAR INFRARED SPECTROSCOPY TO THE ANALYSIS OF WORTS

Transmission near infrared spectroscopy has been evaluated for the analysis of ‘laboratory’ worts. Good agreement between manual and near infrared methods has been obtained for Hot Water Extract and total carbohydrates. Satisfactory calibrations have been developed for fermentable sugars, fermentability and free alpha-amino nitrogen. No sample preparation was required. Hence, this technique provides a rapid and non-destructive method for measuring all these parameters simultaneously.     

Assessment of Spirit Yield in Barley Breeding Lines

A number of random inbred lines of barley, grown at two sites, were assessed for high gravity spirit yield. A range of values was obtained and there was good agreement, for nearly all samples, between high gravity spirit yield and spirit yield predicted from extracts and fermentabilities measured by Institute of Brewing recommended methods. One sample, which gave a low predicted spirit yield but good high gravity spirit yield, was shown to have modified poorly and unevenly. Over both sites, high gravity spirit yield appeared to correlate negatively with grain nitrogen content, but this reflected large differences between sites in grain nitrogen. High gravity spirit yield did not correlate with soluble nitrogen, but did with fermentability, so samples with low levels of fermentability had either low or moderate levels of high gravity spirit yield. It was concluded that identification of genetic factors, influencing extract and fermentability, would enhance breeding of barley cultivars suitable for malt whisky distilling.