ISOLATION AND PARTIAL CHARACTERIZATION OF THE DIMETHYL SULPHIDE PRECURSOR IN GREEN MALT,AND ITS EFFECT ON BEER DIMETHYL SULPHIDE LEVELS

The component from which dimethyl sulphide (DMS) is produced when green malt worts are heated has been purified and its effects on wort and beer DMS levels studied. The green malt precursor can be split into S-methylmethionine and an as-yet uncharacterized ninhydrin-positive component. When the purified precursor is added to worts derived from kilned malts, it is rapidly taken up by yeast, but there is no resulting increase in DMS production. DMS production during fermentation occurs with worts made from kilned malts but not with worts from green malt; however, this difference is not due to an inhibitor being present in green malt worts. The evidence suggests that malt kilning affects the nature as well as the amount of DMS precursor in malts.     

THE MEASUREMENT OF DIMETHYL SULPHIDE PRECURSOR IN MALTS,ADJUNCTS, WORTS AND BEERS

Dimethyl sulphide in beer originates from a precursor in the malt. A method is described for the measurement of this precursor in brewing raw materials, wort and beer. The precursor levels in various green malts, kilned malts, and adjuncts are given. Excess methionine inhibits precursor uptake by yeast during fermentation. Residual precursor is still present in some commercial bottled beers.     

THE RELATIONSHIP OF DIMETHYL SULPHIDE LEVELS IN MALT,WORT AND BEER

The half-life for the conversion of malt dimethyl sulphide (DMS) precursor to free DMS has been determined at various temperatures and pH values. At pH 5·2 the half-life of the precursor in wort (S.G. 1·060) at its boiling point is 38 min, and is doubled for each 6°C fall in temperature. At pH 5·5 the half-life at the boiling point is 32·5 min. Knowing the stability of the precursor at the various temperatures in the brewing process, the extent of conversion to free DMS in wort at pitching can be predicted for malt of a given precursor content and for a given set of process conditions. The results of DMS analyses of samples taken during brewery trials are in reasonable agreement with the predicted values. This work involved infusion mashing only, but the same principles apply to decoction mashing. The fate of precursor and free DMS during fermentation and conditioning has been followed on a production scale. With some brews, where high levels of free DMS were present at pitching, much free DMS was lost during fermentation. Also, precursor DMS reappeared in the beer after a few days and there was some increase in the level of free DMS. The DMS precursor in green malt has been isolated by ion-exchange and gel-filtration chromatography. A preparation has been obtained which has 0·6 mol potential DMS per mol amino nitrogen. Thin layer chromatography showed that the preparation and its hydrolysis product had the same R_f values as S-methylmethionine and homoserine respectively.     

IDENTIFICATION OF THE DIMETHYL SULPHIDE PRECURSOR IN MALT

The heat-labile dimethyl sulphide (DMS) precursor in green malt has been isolated using ion exchange chromatography and gel-filtration under mild conditions. The final preparation had ～0.7 mol potential DMS per mol amino groups. Thin layer chromatography of this preparation, and the gel-filtration behaviour of both crude and purified preparations, showed that the precursor had identical properties to S-methylmethionine (SMM). The DMS precursor in a continental lager malt was also examined. The malt had been kilned under conditions which were expected to lead to a significant content of ‘active’ DMS precursor (reported to be metabolized to free DMS by suitable yeast strains, and contrasting with the ‘inactive’ green malt precursor). The DMS precursor in an extract of this kilned malt had the same elution volume on Sephadex G15 as SMM. There was no evidence for any difference from the DMS precursor in green malt. It is concluded that SMM is the only heat-labile DMS precursor in malt.     

Impact of Dark Specialty Malts on Extract Composition and Wort Fermentation

Dark specialty malts are important ingredients for the production of several beer styles. These malts not only impart colour, flavour and antioxidative activity to wort and beer, they also affect the course of wort fermentations and the production of flavour‐active yeast metabolites. The application of considerable levels of dark malt was found to lower the attenuation, mainly as a result of lower levels of fermentable sugars and amino acids in dark wort samples. In fact, from the darkest caramel malts and from roasted malts, practically no fermentable material can be hydrolysed by pilsner malt enzymes during mashing. Compared to wort brewed with 50% pilsner malt and 50% dark caramel malt or roasted malt, wort brewed with 100% pilsner malt contained nearly twice as much fermentable sugars and amino acids. Reduced levels of yeast nutrients also lowered the fermentation rate, ranging from 1.7°P/day for the reference pilsner wort of 9 EBC to 1.1°P/day for the darkest wort (890 EBC units), brewed with 50% roasted malt. This additionally indicates that lower attenuation values for dark wort are partially due to the inhibitory effects of Maillard compounds on yeast metabolism. The application of dark caramel or roasted malts further led to elevated levels of the vicinal diketones diacetyl and 2,3‐pentanedione. Only large levels of roasted malt gave rise to two significant diacetyl peaks during fermentation. The level of ethyl acetate in beer was inversely related to colour, whereas the level of isoamyl acetate appeared to be affected by the use of roasted malt. With large levels of this malt type, negligible isoamyl acetate was generated during fermentation.     

Brewing with 100% green malt – process development and key quality indicators

Brewing with undried, germinated (green) malt has the potential to lower energy and water usage in the malting and brewing chain. However, doing so introduces technical and biochemical (flavour) challenges. Beers were brewed using 100% green malt (n = 3) or kilned pilsner malt (n = 3), prepared from the same batch in each case, utilising the pilot brewery at KU Leuven (2.5 hL). Three further pairs of beers were brewed whereby the green malt was pre-steeped under deaerated water for 1 hour; this procedure was previously shown to lower LOX activity in green malt. Six green malt beers were brewed with acceptable specifications in terms of pH, alcohol content, foam stability and colour. No significant taints or obvious defects were detected in green malt beers. Increased S-methyl methionine levels were measured in worts and beers made from green malt, however DMS concentrations in the finished beers did not differ significantly from the reference beers. Furthermore, the results demonstrated promising indicators for flavour stability, such as reduced TBI, lower residual FAN and trihydroxy fatty acid (THFA) levels in brews using untreated green malt. Using re-steep water in green malt brewing (for reasons of water economy), however, increased THFA levels, possibly because oxygen uptake was not adequately controlled at this step. Whilst further process optimisations are undoubtedly required, it is shown that an acceptable lager style beer could be brewed to a specification not dissimilar to that of a kilned malt control beer, using 100% green malt with intact rootlets. © 2020 The Authors. Journal of the Institute of Brewing published by John Wiley & Sons Ltd on behalf of The Institute of Brewing & Distilling     

EFFECT OF KILNING ON MALT STARCH AND ON THE DEXTRIN CONTENT OF RESULTING WORTS AND BEERS

Examination of starches separated from green malt and from kilned malts of colour ranging between 3 and 23 shows that only small amounts of “new” carbohydrate linkages are formed in the starches during kilning, though the colour changes from white to buff and the amount of contaminating protein increases. Nevertheless, standard enzymic treatments show that the higher the colour value of the malt, the less susceptible it is to attack by amylases, and the larger is the content of non-fermentable, non-dialysable dextrin. Although the amount of non-dialysable dextrin rises as the colour value of the malt increases, dextrin content of the derived wort is not directly related to the enzymic activity of the malt. Thus, the non-fermentable dextrin of wort from malt of colour 3 is little higher than that of wort from green malt, despite the considerably higher diastatic power of the latter. Head retention values of draught and bottled beers from the green malt and kilned malts of colours 3, 6 and 13 show no correlation with dextrin content.     

Differences in protein content and foaming properties of cloudy beers based on wheat malt content

To investigate differences in protein content, all barley malt beer, wheat/barley malt beer and all wheat malt beer were brewed, and the protein during mashing, wort, fermentation and beer determined. It was shown that protein was mainly extracted during mashing and the protein rest phase, decreased in the early stages of fermentation and remained almost steady during wort boiling and cooling, in the middle and late stages of fermentation. By separating beer foam from beer, similar protein bands of 51.7, 40.0, 27.3, 14.8, 6.5 and < 6.5 kDa appeared in the three beers, defoamed beers and beer foams using the sodium dodecyl sulphate polyacrylamide gel electrophoresis. Quantitatively, protein bands of 6.5–14.8 and <6.5 kDa had the highest contents in the three beers. Unique bands at 34, 29.2, 23.0, 19.7 and 17.7 kDa were found in beer, defoamed beer and beer foam from wheat beer and all‐wheat malt beer, respectively. Wheat beer foam showed the best foam stability and the protein in all barley malt beer showed the best migration to the foam. The beer foam properties were influenced by not only protein content but also protein characteristics and/or origin. It is suggested that the barley malt contributed the beer foam ‘skeleton protein’ while protein components from wheat malt kept the foam stable. © 2018 The Institute of Brewing & Distilling     

The Impact of a Xanthohumol‐Enriched Hop Product on the Behavior of Xanthohumol and Isoxanthohumol in Pale and Dark Beers: A Pilot Scale Approach

Based on the health‐promoting properties of xanthohumol (XN), the production of an enriched beer in this substance would be of interest to the brewing industry, from the perspective of pointing out the benefits that beer could bring consumer health. For that purpose, in this work efforts were applied to produce a beer enriched in XN. Also investigated was the influence of a XN‐enriched hop product on the content of XN and isoxanthohumol (IXN) in pale and dark beers. It was verified that XN was largely converted into IXN during wort boiling. However, the use of dark malts revealed a positive effect on the thermal isomerisation of XN. These results are indicative of the isomerisation‐inhibiting effect of the stout production process, which resulted in high levels of XN in the beer. Further losses of XN were due to incomplete extraction from the hops into the wort, adsorption to insoluble malt proteins and adsorption to yeast cells during fermentation. It was possible to produce a dark beer enriched in XN (3.5 mg/L) by using coloured malt (caramel malt, roasted malt and roasted malt extract) and a special XN hop extract combined with late hop usage during wort boiling.     

TRACE VOLATILE CONSTITUENTS OF BEER DIMETHYL SULPHIDE FORMATION IN MALT

Dimethyl sulphide formation in malt occurs primarily during the kilning process, and is significantly dependent upon the maximum kilning temperature. During mashing a considerable fraction of the dimethyl sulphide of malt is transferred to the wort. Since this is not always completely removed by the wort boiling process, the remaining dimethyl sulphide, although decreased during fermentation, may be sufficient to impart a noticeable dimethyl sulphide aroma to the finished beer.     

CONTROL OF THE DIMETHYL SULPHIDE CONTENT OF BEER BY REGULATION OF THE COPPER BOIL

In pilot brewing trials where wort boil time was varied from 15 min up to 2 hours the DMS contents of the resultant beers were greatly altered without substantial effect on many other aspects of beer quality. The worts were separately fermented by two yeast strains and a consistent difference between DMS levels of the pairs of beers was found which was almost independent of the boil time. The breakdown of the malt-derived DMS precursor in boiling wort was found to be a first order reaction having a rate constant of about 0.02 min^?1, implying a half life of about 35 min. The breakdown rate of S-methylmethionine in a boiling sucrose solution at pH 5.3 was similar. Free DMS was found to leave this wort substitute rapidly, again as a first order process, but with a rate constant of 0.18 min^?1 (t_1/2 ～ 4 min) when the evaporation rate was 17%/h.     

啤酒酿造中二甲基硫、甲基蛋氨酸、二甲基亚砜含量的变化与控制

为控制啤酒风味物质二甲基硫（dimethyl sulfide,DMS）的含量,实验运用气相色谱-氢火焰离子化检测器测定了15 种国内市售啤酒样品DMS的含量,并研究了啤酒酿造中DMS、甲基蛋氨酸（S-methylmethionine,SMM）、二甲基亚砜（dimethyl sulfoxide,DMSO）的含量变化。结果表明,抽取的国内市售啤酒样品中有47%的样品DMS含量超出60 μg/kg,特别是爱尔啤酒（Ale）中71%的样品DMS含量超出60 μg/kg,最高值达到105 μg/kg;实验抽检10 种国内市售麦芽DMS、SMM、DMSO含量,不同麦芽间DMS与二甲基硫前驱体（dimethyl sulfide precursor,DMSP）差异较大,差异最高达160%,且多数小麦芽的DMS与DMSP含量低于澳麦芽。实验研究了麦汁煮沸、发酵过程中的DMS及DMSP含量的变化,在100 ℃、pH 5.6时,麦汁煮沸40 min后DMS含量比初始含量降低92%,SMM的半衰期为31 min。发酵过程中DMS含量在发酵旺盛期时有明显的下降,最高下降41%。     

THE MEASUREMENT OF DIMETHYL SULPHOXIDE IN BARLEY AND MALT AND ITS REDUCTION TO DIMETHYL SULPHIDE BY YEAST

Dimethyl sulphoxide (DMSO) is a normal component of malt and barley. A method is described for its extraction and estimation. DMSO is produced by the oxidation of dimethyl sulphide (DMS), particularly during kilning of malt, and higher levels are found in malts subjected to ale kilning schedules. DMS may also be oxidized during wort preparation. DMSO can be reduced to DMS by yeast in glucose/salts medium, by yeast cell suspensions and by a cell-free extract. Reduction of DMSO is inhibited by methionine sulphoxide. The results suggest that reduction of DMSO may account for the DMS produced during fermentation of ale and lager worts.     

MALTING AND BREWING WITH BARLEYS HAVING BLUE ALEURONES

Blue aleurones in barley are associated with elevated levels of polyphenolic materials such as anthocyanins and anthocyanidins. A rapid method has been developed for assessing the anthocyanin content of barleys, malts and worts. Malts were prepared from a range of barleys, some of which had intensely blue aleurones, while others were only slightly blue or showed no visible pigmentation. The malting quality of barley was not affected by aleurone colour and ales and lagers of sound flavour as well as acceptable analytical parameters were brewed from malts having pronounced blue aleurones. In some cases sweet worts prepared from blue aleurone malts had a slight pinkish tinge, but this disappeared during wort boiling, and beer colours were normal. Levels of anthocyanins in barley correlated with those in malt and in wort. However, the concentration of anthocyanins was unrelated to the amount of anthocyanogens or total polyphenols. High anthocyanin levels in either barley or beer had no deleterious effects on beer flavour or the rate of haze development.     

Roasting of malt and xanthohumol enrichment in beer

The use of commercial roasted malts increases the content of the hop polyphenol xanthohumol (XN) in beer. This carrier effect is caused by high molecular melanoidins from roasted malt. Three roasting trials with different malts were performed in order to study the development of XN enrichment of wort and beer in laboratory and brewing trials. Different colour measurements, malt and flavour analysis, radical formation and antioxidative activity of selected samples were carried out. Furthermore, sensory tests of beers were conducted. During roasting the XN and isoxanthohumol enrichment in wort and beer rose with the roasting intensity of malt until it reached a maximum. The XN content in wort increased linearly with the colour of wort made from the malt samples. In PVPP-treated worts and in filtered beers, the XN content increased exponentially with the colour. After passing through a maximum, both the colour value and XN content in wort and beer decreased. Interestingly, the colour losses were more intensive than the losses of XN in worts and beers. The development of radical formation and the reducing power was linked during roasting. That means reducing groups of melanoidins are responsible for reducing power and prooxidative properties of malts. These functional groups of melanoidins are involved in the development of XN enrichment properties, because a linear correlation between these parameters was found. In conclusion, the roasting regimes showed potential for the development of special malt for the XN enrichment in beer- or malt-based beverages in late roasting stages. The use of this special malt brings more XN with less coloured malt in beer.     

100%大麦啤酒酿造过程中老化Strecker醛的研究

分别采用上面发酵工艺与下面发酵工艺进行100%大麦啤酒及100%麦芽啤酒的酿制,并对其麦汁的氨基酸含量、老化Strecker醛、自由基以及新鲜啤酒中老化Strecker醛的含量等进行了对比分析。研究发现,就麦汁而言,100%大麦麦汁中老化Strecker醛的含量都明显低于100%麦芽麦汁;同样的麦汁,上面发酵方式还原Strecker醛的能力明显优于下面发酵方式。就啤酒而言,经酵母还原后,新鲜啤酒中的老化Strecker醛含量较麦汁含量低,且100%大麦啤酒中老化Strecker醛的含量低于100%麦芽啤酒中的含量。100%麦芽麦汁的自由基含量是100%大麦麦汁的近3倍。这都预示着100%大麦啤酒的风味稳定性（新鲜度）明显好于100%麦芽啤酒。     

Corn grist adjunct – application and influence on the brewing process and beer quality

In the brewing industry, barley malt is often partially replaced with adjuncts (unmalted barley, wheat, rice, sorghum and corn in different forms). It is crucial, however, to preserve constant quality in the beer to meet the expectations of consumers. In this work, how the addition of corn grist (10 and 20%) influences the quality of wort and beer was examined. The following parameters were analysed: wort colour, dimethyl sulphide (DMS) and protein content, non‐fermentable extract, extract drop during fermentation, alcohol content and the attenuation level of the beer, together with filtration performance. The samples (all‐malt, and adjunct at 10 and 20% corn grist) were industrial worts and the beers produced in a commercial brewery (3000 hL fermentation tanks). The application of 10 and 20% corn grist had an effect on the wort colour, making it slightly lighter (11.1 and 10.5°EBC, respectively) than the reference barley malt wort (12.2°EBC). The free amino nitrogen level, DMS and non‐fermentable extract were significantly lower in the worts produced with the adjunct; the alcohol content and attenuation levels were higher in the beers produced with adjunct. The use of corn grist, at the level of up to 20% of total load, appears to affect some of the technological aspects of wort and beer production, but it does not significantly influence the final product characteristics. Copyright © 2014 The Institute of Brewing & Distilling     

Yeast Proteolytic Activity During High and Low Gravity Wort Fermentations and its Effect on Head Retention

The aim was to discover the effect of high gravity brewing on yeast protease activity during fermentation, on the loss of hydrophobic polypeptides from wort during fermentation, and on the foam stability of stored beer. The hydrophobic polypeptide content of low (10° Plato) gravity worts showed a steady decline throughout fermentation, but for the 20° Plato wort there was a rapid decline over the first 8 days of fermentation, followed by little change over the remaining period. The decrease in hydrophobic polypeptides was greater in the high gravity fermentation. Proteinase A increased during fermentations with the highest levels being present at the end of fermentations. High gravity fermentations exhibited levels of yeast protease that from the 3rd to 11th day of fermentation were at least twice the values of the low gravity fermentations. The high gravity brewed beer contained significantly higher levels of proteinase A activity than the low gravity brewed beer. The inclusion of FERMCAP™, an antifoam, in high gravity wort did not affect either the hydrophobic polypeptide levels or foam stability of the resultant beer. This suggests that proteinase A, rather than fermenter foaming, must be the major contributor to the lack of foam stability of high gravity brewed beer. Head retention measurements conducted on the high and low gravity brewed bottled beers, over a five month period, demonstrated a steady decline in foam stability for both beers. The declines in head retention did not occur in high and low gravity beers that had been pasteurised.     

FLAVOUR AND HAZE STABILITY DIFFERENCES DUE TO HOP TANNINS IN ALL-MALT PILSNER BEERS BREWED WITH PROANTHOCYANIDIN-FREE MALT

All-malt Pilsner beers were brewed with proanthocyanidin-free malt (ant 13 × 13 × Rupal). Hopping was with n-hexane tannin-free hop extract with or without n-hexane extracted hop residue or with whole leaf hops. The different beers were analysed chemically and presented to an expert panel to detect possible preferences and differences in bitterness and astringency. The impact of hop proantho-cyanidins on haze formation is comparable to that of malt proanthocyanidins. No differences between the beers were found in triangular tests. Statistical analysis of the paired comparison tests showed that hop proanthocyanidins do not contribute to the bitterness of beer since beer brewed with tannin-free hop extract is slightly more bitter than beer brewed with whole leaf hops. No differences in astringency were noted between beers brewed with or without hop proanthocyanidins and it was found that the panel expressed a very slight preference for beer brewed with n-hexane hop extract over beer brewed with both n-hexane hop extract and extracted hop residue.     

Enrichment of xanthohumol in the brewing process

Xanthohumol (XN), a component of hops, is lost in significant quantities in the conventional brewing process. In commercial beers less than 0.2 mg XN/L are found. In order to increase the yield of XN in the brewing process, the parameters of XN recovery were studied. During wort boiling, XN is largely isomerised to isoxanthohumol. Further losses are owing to the precipitation and absorption of XN to yeast cells and haze particles and by filtration. The use of XN-enriched hop products combined with a late hop dosage during wort boiling proved to be effective in increasing the XN content in beer. The yield was further raised by a low-pitching rate and the abnegation of beer stabilisation. The use of dark malts had a positive effect on the XN recovery. Investigations of roasted malt extracts revealed several high-molecular substances that are able to form complexes with XN. These complexes proved to be stable in the brewing process. Depending on the addition of roasted malt or special XN-enriched roasted malt extracts, dark beers with more than 10 mg XN/L were achieved. Results obtained led to a brewing technology that produced on an industrial scale pale wheat beer with more than 1 mg XN/L.