Optimisation of High Gravity and Diet Beer Production in a German Brewery by Flow Cytometry

Increasing the quantity of beer production without diminishing the quality of the product is a key concern of the beer producing industry. Modifications to the brewery's equipment and settings are the most commonly used methods to improve the brewing process, while the supreme importance of the physiological state of the beer producing organisms, the yeast cells, for the productivity of the brewing process is often poorly recognised. The work described here was designed to optimise two processes: the inoculation regime used to produce high gravity bottom-fermenting beer, and the production of high quality diet beer. To achieve these aims, flow cytometry was used to follow changes in the distribution of DNA, neutral lipid and 3β-hydroxsterol contents in Saccharomyces carlsbergensis strains during inoculation, fermentation and storage. This allowed potential time-saving alterations in the process to be identified. Double staining techniques proved that vigorous fermentative activity and long-term survival capacity during main and secondary fermentation requires intense multiplication of the yeast cells during inoculation. The production of high gravity beer was then enhanced by altering the schedule of the wort additions, and thus increasing the yeast's activities related to multiplication. To produce diet beer, oligosaccharides that remain after the standard brewing process are degraded by adding small amounts of wort, usually during secondary fermentation. However, during this period of fermentation the physiological activity of the yeast cells is hampered by low carbon and high ethanol concentrations. Adding small batches of wort at carefully defined time points and in optimised amounts, even during the main fermentation, improves the physiological state of the yeast cells and rapidly decreases the carbon concentration within the fermentation tank. Both of these factors help to promote quick fermentation to a high quality diet beer. Thus, the flow cytometric investigations provided a reliable basis for identifying effective means of improving the process regime for brewing both of these products.     

基于人工神经网络模拟啤酒酿造过程中糖度及乙醇浓度的变化

建立BP神经网络模型模拟啤酒酿造过程中糖度变化和乙醇浓度变化.将啤酒酿造过程中的发酵温度、麦汁浓度、接种量及发酵时间作为输入数据,将糖度变化和乙醇浓度的变化作为输出数据,运用BP神经网络建立啤酒酿造过程的模型.使用此模型模拟了主酵温度8℃、麦汁浓度11°P、接种量为2×107个/mL时糖度变化和乙醇浓度变化,结果糖度预测的均方根误差为2.66％,乙醇浓度预测的均方根误差为14.60％.结果表明,使用此模型能够准确预测啤酒酿造过程糖度变化和乙醇浓度的变化.     

SOME FACTORS AFFECTING THE CONCENTRATION OF DIACETYL IN BEER

Investigations of factors affecting the production of diacetyl during fermentation have shown large differences in the abilities of different strains of brewer's yeast to produce diacetyl. Studies on pilot scale and on small laboratory fermentations have established a connection between the concentration of amino acids in the wort and that of diacetyl in the fully fermented beer. Control of the ratio of amino acids to sugars in wort is thus an important factor in maintaining concentrations of diacetyl in beer at an acceptable level. The effects of increased pitching rate and of high-temperature fermentation in enhancing the production of diacetyl are demonstrated.     

HORACE BROWN MEMORIAL LECTURE THE ROLE OF NITROGEN IN BREWING

The development of chromatographic methods of analysis in the 1940's and 1950's allowed detailed investigations to be made into the role of nitrogenous compounds in malting and brewing. The development of individual amino acids during malting, their extraction in the brewhouse and assimilation by culture yeasts during fermentation are reviewed as are their effect on the quality of the final beer.     

The Horace Brown Medal Lecture: Forty Years of Brewing Research

Horace Brown spent fifty years conducting brewing research in Burton‐on‐Trent, Dublin and London. His contributions were remarkable and his focus was to solve practical brewing problems by employing and developing fundamental scientific principles. He studied all aspects of the brewing process including raw materials, wort preparation, fermentation, yeast and beer stability. As a number of previous presenters of the Horace Brown Lecture have discussed Brown's achievements in detail, the focus of this paper is a review of the brewing research that has been conducted by the author and his colleagues during the past forty years. Similar to Horace Brown, fundamental research has been employed to solve brewing problems. Research studies that are discussed in this review paper include reasons for premature flocculation of ale strains resulting in wort underattenuation including mechanisms of co‐flocculation and pure strain flocculation, storage procedures for yeast cultures prior to propagation, studies on the genetic manipulation of brewer's yeast strains with an emphasis on the FLO1 gene, spheroplast fusion and the respiratory deficient (petite) mutation, the uptake and metabolism of wort sugars and amino acids, the influence of wort density on fermentation characteristics and beer flavour and stability, and finally, the contribution that high gravity brewing has on brewing capacity, fermentation efficiency and beer quality and stability.     

Production of Beer with a Genetically Engineered Strain of S. cerevisiae with Modified Beta Glucanase Expression

This study used a recombinant Saccharomyces cerevisiae strain, which expressed both β‐glucanase enzyme and reduced Pro‐teinase A expression during wort fermentations. The genetic stability and fermentation features of the strain were examined. The recombinant strain's proteinase A activity was reduced compared to the parent strain; β‐glucanase was produced throughout the fermentation. The fermentation with the recombinant S. cerevisiae strain exhibited a larger reduction in β‐glucan content than what was observed with the control strain, with β‐glucan degradation above 80%. The foam stability period was reduced when the beer produced by the recombinant S. cerevisiae was stored for 3 months. SDS‐PAGE analysis of the beer proteins indicated that lipid transfer protein 1 had disappeared. Fermentation studies indicated that based on the parameters examined, this recombinant strain was suitable for industrial beer production.     

Impact of the Long‐Term Maintenance Method of Brewer's Yeast on Fermentation Course,Yeast Vitality and Beer Characteristics

The effect of the long‐term maintenance method used with a brewer's yeast on its technological properties was determined in laboratory fermentation trials with a 12°P all‐malt wort. The trials were performed at a constant temperature and under conditions of constant substrate concentration. Two cultures of a bottom fermenting yeast, Saccharomyces pastorianus RIBM 95, were tested — one culture was maintained by subculturing on wort agar slopes at 4°C and the other culture underwent a three year storage in liquid nitrogen at minus 196°C. Parameters under investigation included yeast vitality measured as acidification power (AP), fermentation time needed to reach an alcohol level of 4%, the yeast cell count, sedimentation of the yeast during the fermentation, and the production of beer flavour compounds in green beer. The yeast culture stored for three years in liquid nitrogen displayed a higher count of suspended cells, required a shorter time to attenuate the wort to produce 4% alcohol and produced a 1.5 to 2.5‐fold higher concentration of a number of flavour compounds. The long‐term storage method did not affect the sedimentation ability and vitality of the yeast strain tested.     

HORACE BROWN MEMORIAL LECTURE A REALISTIC VIEW ON THE ROLE OF RESEARCH IN THE BREWING INDUSTRY TODAY

Present-day economic pressures stimulate technological innovations toward more rapid processing and to minimise both capital and production costs. Fermentation and maturation practices in most production plants are essentially unchanged from the low productivity batch processes. In terms of process economics two alternatives exist—moving to greater volume accelerated-batch processing and applying fully continuous high rate fermentation systems. Central to these issues are the growth controlling aspects of yeast performance in brewery fermentations. The importance of dissolved oxygen on lipid metabolism, yeast growth and beer flavour, the precise role of storage carbohydrates and the critical nature of the glycolytic/gluconeogenic junction in controlling fermentation performance are reviewed. The objectives of process optimisation are to build up sufficient yeast concentrations to gain optimal rate and extent of attenuation and desired flavour development whilst balancing but not over-expending wort nutrients for growth and maintenance. In this regard great promise may lie with new technologies including yeast oxygenation and immobilised cell systems. Particular emphasis is placed upon the ways in which this may be technically achieved and attention is drawn to the complexity of innovation management. Practical implications of factors benefiting and adversely affecting fermentation efficiency and beer flavour in both traditional and continuous immobilised cell systems are discussed.     

Predicting acetic acid content in the final beer using neural networks and support vector machine

Most acetic acid found in beer is produced by yeast during fermentation. It contributes significantly to beer taste, especially when its content is higher than the taste threshold in beer. Therefore, the control of its content is very important to maintain consistent beer quality. In this study, artificial neural networks and support vector machine (SVM) were applied to predict acetic acid content at the end of a commercial‐scale beer fermentation. Relationships between beer fermentation process parameters and the acetic acid level in the fermented wort (beer) were modelled by partial least squares (PLS) regression, back‐propagation neural network (BP‐NN), radial basis function neural network (RBF‐NN) and least squares‐support vector machine (LS‐SVM). The data used in this study were collected from 146 production batches of the same beer brand. For predicting acetic acid content, LS‐SVM and RBF‐NN were found to be better than BP‐NN and PLS. For the comparison of RBF‐NN and LS‐SVM, RBF‐NN had a better reliability of model, but lower reliability of prediction. SVM had better generalization, but lower reliability of model. In summary, LS‐SVM was better than RBF‐NN modelling for the prediction of acetic acid content during the commercial beer fermentation in this study. Copyright © 2013 The Institute of Brewing & Distilling     

啤酒酵母在红薯淀粉麦芽汁中的发酵特性研究

本实验研究了酵母在红薯淀粉做辅料的麦汁中的发酵特性,并与其在大米做辅料的麦汁中的发酵特性作了对比,结果表明其完成主发酵所用的时间、可发酵浸出物的变化、还原糖含量的变化、α-氨基氮的利用情况、双乙酰的生成量等均无明显的区别,而且酵母的生长繁殖正常。红薯淀粉作辅料的啤酒色度较大。     

薏米啤酒的加工工艺研究与优化

以薏米为试验材料,采用响应面法对薏米啤酒的工艺参数进行研究和优化。在单因素试验基础上,研究料水比、薏麦比、发酵温度对薏米啤酒风味的影响,以薏米啤酒感官评分为响应值,利用Box-Behnken中心组合试验设计建立数学模型。研究结果表明,优化后的工艺参数为:料水比1︰4.76(g/mL)、薏麦比0.2︰1(g/g)、发酵温度10.64℃。通过黄金分割法,确定卡拉胶添加量为31.12 mg/L时澄清效果最佳。     

THE INFLUENCE OF 2-ACETOHYDROXY ACIDS ON THE DETERMINATION OF VICINAL DIKETONES IN BEER AND DURING FERMENTATION

During sampling and determination of diacetyl, 2-acetohydroxy acids are easily converted to vicinal diketones. A simple procedure for gas chromatographic determination of diacetyl, 2-acetolactate, acetoin and the homologous compounds is given. By careful sampling, less than 0·01 ppm of diacetyl was detected during the main fermentation in one brewery, whereas another strain of brewer's yeast yielded a maximum of 1·7 ppm of diacetyl. When samples of fermenting liquids are exposed to air at 60°C, complete conversion of 2-acetohydroxy acids takes place in less than one hour. The possibility that part of 2-acetolactate may be converted to acetoin, however, cannot be excluded. In finished beer 2-acetolactate levels of 0·2–0·5 ppm were observed. During the main fermentation the values ranged from 0·5–2·5 ppm.     

A RAPID PROCEDURE FOR REDUCING THE DIACETYL CONTENT OF BEER*

Beer produced by rapid fermentation procedures can contain a substantial quantity of precursor substances which give rise to diacetyl and 2,3-pentanedione when the beer is stored. A conditioning unit has been developed which rapidly converts these precursors to the free diketones, which are then removed by yeast. Maturation of the beer is thus achieved at a rate similar to that of the initial fermentation.     

THE CAMBRIDGE PRIZE LECTURE 1996 FROM FIELD TO FIRKIN: AN INTEGRATED APPROACH TO BEER CLARIFICATION AND QUALITY

A concerted and integrated research programme has considerably enhanced the current understanding of fining agents and their effect on beer clarification and quality. In addition to previously published data, much new information has been presented to help develop this understanding. The numerous complex and often inter-related factors that affect both isinglass and copper fining performance have been discussed both qualitatively and quantitatively. This information has been used to identify the following critical control parameters which will facilitate the optimisation of beer quality and brewing process efficiency : Pre- and post-fining beer temperature beer particle levels and NMP charge wort pH cold wort clarity beer pH pre-filtration beer clarity . The discrete unit operations of the brewing process have been integrated into a single continuous process, where the efficiency of each operation depends on the efficiency of all of the previous operations, and impacts directly on that of all subsequent operations. Using this approach, beer clarity problems can in certain circumstances be traced back to variations in the microflora present on barley in the farmer's field. Thus to truly control beer quality, the brewing process must be considered to start not with mashing or malting, but rather with barley growing, and the process must be managed right through “from field to firkin” .     

THE CONTROL OF VOLATILE ESTER SYNTHESIS DURING THE FERMENTATION OF WORT OF HIGH SPECIFIC GRAVITY

Some effects have been studied of fermenting wort of unusually high specific gravity, followed by dilution with water to give beer of normal original gravity. This procedure permits increased overall rates of beer production, but matching of flavours requires control of the level of flavour determinants. If not controlled, the concentration of volatile esters may be disproportionately increased so that, after dilution, beer flavour is markedly different. Higher alcohol production is not affected in this way. The concentration of esters can be adjusted to appropriate levels by increasing the production of yeast mass during fermentation. Unsaturated fatty acids, which increase yeast dry matter production without altering the rate of fermentation, are particularly effective in reducing the extent of ester synthesis.     

Improving acetic acid production of Acetobacter pasteurianus AC2005 in hawthorn vinegar fermentation by using beer for seed culture

Fruit vinegar can be obtained biologically from the fermentation of fruit wine using species of the genera Acetobacter. However, the productivity of vinegar is generally low because of the inhibition by substrate ethanol and product acetic acid. In this research, the yield of acetic acid, as well as the cell growth, in hawthorn vinegar fermentation was enhanced using beer for seed preparation. The higher yield of acetic acid of AC2005 was because of the higher alcohol dehydrogenase and aldehyde dehydrogenase activities caused by using beer for seed preparation. Furthermore, Acetobacter pasteurianus AC2005 showed acid stability during the fed‐batch fermentation of hawthorn vinegar.     

HORACE BROWN MEMORIAL LECTURE A CONSPECTUS OF BREWING PROGRESS

Horace Brown, who started as a brewer just over a hundred years ago, was an outstanding pioneer in the introduction of science to explain and guide brewing practice. His own and Pasteur's work saved brewing from repeated crises due to spoilage infections and so made it safe to brew thoughout the year. Attempts to control infection gave rise to Brown's great interest in the nitrogen compounds and led him to notable discoveries, apart from his outstanding work in many other fields. The writer followed with researches on the nitrogen compounds of barley, malt, wort and beer. These are summarized here and the potentialities for further developments today in the study of proteins are outlined. Against the background of the changing scene in brewing, a summary is given of the writer's further work on the agricultural aspects of barley growing, on barley germination and malting and on “sediment action” and yeast nutrition in fermentation. In addition a summary is given of his contributions in biological engineering, to continuous fermentation and to a scientifically-based conditioning process, using lupulin extracted from hops and incoporating a greatly accelerated haze-prevention method, which is economical. While drastically shortening the process, this and the other stages have been beneficial rather than otherwise to beer flavour.     

THE EFFECT OF CARBOHYDRATE ADJUNCTS ON BREWER'S WORT FERMENTATION BY SACCHAROMYCES UVARUM (CARLSBERGENSIS)*

The use of carbohydrate adjuncts such as sucrose, fructose and glucose in brewer's wort significantly modifies the initial wort sugar spectrum and also the pattern of sugar uptake during fermentation by a strain of Saccharomyces uvarum (carlsbergensis). Under these conditions, the concentration of glucose and fructose in the wort was observed to increase when compared to worts in which corn starch was employed as an adjunct and glucose was taken up at a faster rate than fructose. The increase in glucose concentration in the wort also resulted in severe repression of maltose and maltotriose utilization with significant levels of these sugars remaining in the beer produced.     

Current Developments in Malting and Brewing Trials with Sorghum in Nigeria: A Review

Initially, large‐scale lager beer brewing with sorghum malts proved highly intractable due to a number of biochemical problems including: high malting losses estimated at 10–30% as against 8–10% for barley; high gelatinisation temperatures which limited starch solubilisation/ hydrolysis by the amylolytic enzymes during mashing; low extract yield/low diastatic power (DP) due to inadequate hydrolytic enzyme activities especially β‐amylase; low free α‐amino nitrogen (FAN) due to inadequate proteolysis limiting yeast growth during fermentation; high wort viscosities/beer filtration problems due to low endo‐β‐1,3; 1–4‐glucanase activities on the endosperm cell walls causing the release of some β‐glucans. Strident research efforts using improved Nigerian sorghum malt varieties (SK5912, KSV8 and ICSV400) have reported some encouraging results. The knowledge of the biochemical integrity of the endo‐β‐glucanases of the sorghum malt is helping to elucidate their mode of activity in the depolymerisation of the β‐glucans. This is bound to ensure process efficiency in sorghum beer brewing, reduce beer production costs and ultimately, produce a Pilsner‐type of lager beer with 100% sorghum malt.