Degradation of a Foam‐Promoting Barley Protein by a Proteinase from Brewing Yeast

The degradation of a major protein component in beer, the lipid transfer protein (LTP1), by the yeast proteinase A was determined. Another major protein fraction in beer, the protein Z‐fraction, was not degraded by this enzyme. Protein preparations from beer and barley containing LTP1 were examined for degradation by proteinase A using SDS‐PAGE, immunoblotting and RP‐HPLC. LTP1 from barley was completely resistant to proteinase A, whereas LTP1 concentrated from beer was cleaved. We conclude that LTP1 was modified during the brewing process, thus rendering it more susceptible to proteinase A degradation.     

Malting Barley Grain Non‐specific Lipid‐Transfer Protein (ns‐LTP): Importance for Grain Protection

A basic protein (pI < 9) was isolated to homogeneity from a domestic cultivar of malting barley grain (Hordeum vulgare). In its unreduced form it exists as a dimer of a 9 kDa protomer with four disulphide bridges. These characteristics together with protein sequence data revealed that the isolated protein belongs to the class of ns‐LTP. The antifungal potency of malting barley grain ns‐LTP was examined on Saccharomyces cerevisiae, Candida albicans and the plant pathogen Fusarium solani growth in vitro. It was found that ns‐LTP inhibits Saccharomyces and Fusarium growth; the concentration required for 50% inhibition after 24 h of incubation (IC₅₀) was 100 and 80 μg/mL, respectively. On the basis of these results, the importance of ns‐LTP for barley grain protection from fungal diseases has been discussed.     

Effects of Lipid‐Transfer Protein from Malting Barley Grain on Brewers Yeast Fermentation

A lipid‐transfer protein (LTP), which belongs to a family of pathogenesis‐related (PR) proteins, was isolated from malting barley grain. This LTP significantly decreased fermentation and respiration of brewers yeast (Saccharomyces cerevisiae) and caused the leakage of cell constituents. These effects were dose dependent tending to saturation at higher concentrations (～ 200 μg/mg yeast dry weight cells). It was found that LTP survives the thermal treatment during the mashing process. Despite yeast fermentation inhibition in vitro, this LTP did not appear to cause impairment of yeast fermentation capability in the brewing process.     

Utilization of the Linear Mode of MALDI‐TOF Mass Spectrometry in the Study of Glycation During the Malting Process

The process of glycation during the malting process was monitored by the linear mode of matrix‐assisted laser desorption/ionization mass spectrometry (MALDI‐TOF MS). Water‐soluble proteins were investigated and two hulled barley varieties, Jersey and Tolar, were compared to the hulless line KM 1910. The crude extracts of the proteins obtained from the grain, the malt, and aliquots collected every 24 h during the malting process, were mixed with the matrix (2,6‐dihydroxyacetophenone) and analyzed by mass spectrometry. The protein composition of the barley changed during the malting process. The protein patterns did not differ significantly between the three varieties of the barley grains. However, significant differences between the malts were evident. Results showed the influence of the malting process on the glycation of certain water‐soluble barley proteins, nonspecific lipid transfer protein 1 (LTP1) and protein Z, of which the glycated forms survived the brewing process. These major barley proteins are very important for the formation and stability of beer foam and glycation may prevent their precipitation. Analysis results indicated that slight glycation of the proteins had occurred on the second day of malting. The linear mode of MALDI‐TOF mass spectrometry was used as a fast and simple method for monitoring the patterns of low‐molecular weight barley proteins with regard to barley variety discrimination. This procedure also enables the selection of barley varieties suitable for the malting industry.     

A Review: The Role of Barley Seed Pathogenesis‐Related Proteins (PRs) in Beer Production

Pathogenesis‐related proteins (PRs) are involved in the protection of dormant and germinating cereal seeds against pathogenic microorganisms and pests. Moreover, barley grain PRs have considerable technological importance in the brewing of beer. The compact structure of PRs allows them to survive the hostile conditions of the extracellular compartments, where they are usually localized, and enables the endurance of certain classes of PRs during the harsh conditions of the technological steps of beer production. Beer proteome maps reveal a dominant presence of PRs and facilitate association between particular proteins and specific beer quality traits. Current knowledge on the influence of PRs on the various aspects of beer quality has been summarized in this review.     

Multivariate analysis for feasibility of Korean six‐row barleys for beer brewing

The feasibility of using six‐row barley, which is produced more often than two‐row barley (malting barley) in Korea, for beer brewing was studied. Beer was brewed from one variety of two‐row barley (Jinyang, malting barley) and four varieties of six‐row barley (Jasujungchal and Hinchalssal which are unhulled; Dahyang and Samgwangchal which are hulled). Using principal component analysis of the material properties in malting, mashing and fermentation, and the sensory properties of beer, the barley was categorized into three groups: group 1 (Jinyang and Dahyang), group 2 (Samgwangchal and Hinchalssal) and group 3 (Jasujungchal). Group 1 was distinctive for extract (dry basis), Brix and carbonation; group 2 was characterized by alcohol, foam stability and sour odour; and group 3 was characterized by malt protein and sour taste. The brewing qualities of group 1 were superior to those of the other groups. Among the Korean six‐row barley varieties, Dahyang was found to be the most suitable for beer production. Copyright © 2014 The Institute of Brewing & Distilling     

125th Anniversary Review: The science of the tropical cereals sorghum,maize and rice in relation to lager beer brewing

Mainstream lager beer brewing using the tropical cereals sorghum, maize and rice, either as malt or as raw grain plus commercial enzymes, is becoming widespread. This review examines the differences in composition between these tropical cereals and barley and their impact on brewing processes and beer quality. All of these cereals have a starch gelatinization temperature some 10 °C higher than barley. The sorghum prolamin proteins are particularly resistant to proteolysis owing to disulphide cross‐linking involving γ‐kafirin. Unlike barley, the major endosperm cell wall components in sorghum and maize are arabinoxylans, which persist during malting. The rice cell walls also seem to contain pectic substances. Notably, certain sorghum varieties, the tannin‐type sorghums, contain considerable levels of condensed tannins (proanthocyanidins), which can substantially inhibit amylases, and probably also other brewing enzymes. Tropical cereal malts exhibit a similar complement of enzymic activities to barley malt, with the notable exception of β‐amylase, which is much lower and essentially is absent in their raw grain. Concerning beer flavour, it is probable that condensed tannins, where present in sorghum, could contribute to bitterness and astringency. The compound 2‐acetyl‐1‐pyrroline, responsible for the popcorn aroma of maize and also the major aroma compound in rice, presumably affects beer flavour. However, much more research is needed into tropical cereals and beer flavour. Other future directions should include improving hydrolysis of prolamins into free amino nitrogen, possibly using prolyl carboxypeptidases and investigating tropical cereal lines with useful novel traits such as high amylopectin, high protein digestibility and low phytate. Copyright © 2013 The Institute of Brewing & Distilling     

Breeding of lipoxygenase‐1‐less malting barley variety ‘Satuiku 2 go’

The lipoxygenase‐1‐less (LOX‐less) trait has positive effects on beer quality, in particular, improvement of flavour stability related to the reduction of beer‐deteriorating substances such as trans‐2‐nonenal. ‘Ryohfu’ is the only spring‐sown malting barley variety grown in Hokkaido, located in the northern part of Japan, and has been used in the Japanese brewing industry for over 20 years. ‘Satuiku 2 go’ was developed as the first LOX‐less malting barley variety in Japan by successive back‐crossing with molecular marker‐assisted selection to introduce the LOX‐less trait into the recurrent parent ‘Ryohfu’. The agronomic performance and general malt quality of ‘Satuiku 2 go’ were almost equivalent to those of ‘Ryohfu’. Wort and beer analyses at the pilot‐scale brewing trial indicated that the LOX‐less trait had little effect on the general characteristics. In contrast, the beers made from ‘Satuiku 2 go’ malt exhibited reduced levels of trans‐2‐nonenal and trihydroxyoctadecenoic acid. The sensory evaluation demonstrated the superiority of ‘Satuiku 2 go’ beers stored under differing conditions in terms of staleness. It can be concluded that the LOX‐less trait was effective in different genetic backgrounds of the recurrent parents used for the development of LOX‐less malting barley varieties. Copyright © 2018 The Institute of Brewing & Distilling     

啤酒酿造过程中脂转移蛋白糖基化形式的检出

利用MALDI TOF生物质谱检测出了啤酒中LTP的糖基化形式,肯定了啤酒中糖蛋白的存在。LTP糖基化形式存在于从麦芽到啤酒泡沫的整个啤酒酿造过程。酿造过程中蛋白质糖基化的发生增加了蛋白的亲水性,从侧面解释了LTP等强疏水性蛋白稳定存在于啤酒中的原因。     

The influence of LOX‐less barley malt on the flavour stability of wort and beer

The aim of this study was to investigate the influence of lipoxygenase‐less (LOX‐less) barley malt on the quality of wort and beer, with the main focus on beer flavour stability. In the current study, pilot‐scale (1000 L) brewing trials were conducted with a control barley malt AC Metcalfe and a LOX‐less barley malt, PolarStar. The results clearly indicated that the LOX‐less barley malt showed less nonenal potential than the control, although LOX activities in both barley malts were relatively low. The beer brewed from the LOX‐less barley malt contained much lower concentrations of trans‐2‐nonenal (T2N) and gamma‐nonalactone, especially after the (forced or natural) aging of the beer, compared with the beer brewed under the same conditions using the control malt. The sensory panel evaluation indicated similar results in the general flavour profile. The freshness scores of beer brewed from the LOX‐less malt were higher than those from the control malt, and this was more pronounced after forced aging. In addition, the beer brewed from LOX‐less malt had a much better foam stability, almost 30 s (NIBEM test). These results confirm that the use of the LOX‐less barley malt was beneficial to beer flavour stability and foam stability. Copyright © 2014 The Institute of Brewing & Distilling     

PROANTHOCYANIDIN-FREE BARLEY—MALTING AND BREWING

Colloidal haze in beer is due to the precipitation of proteins by proanthocyanidins. Carlsberg Research Center reported in 1977 that the use of barley mutants which block the biosynthesis of catechins and proanthocyanidins in the grain prevents the formation of haze in beer. The results from ten years of malting and brewing research with proanthocyanidin-free barley and malt are reviewed.     

Construction of recombinant industrial <Emphasis Type="Italic">S. cerevisiae</Emphasis> strain with barley lipid-transfer protein 1 secretion capability and lower PrA activity

Beer barley LTP1 in beer is an important component of beer foam, and it participates in the formation of beer foam. The digestion of beer barley LTP1 by proteinase A from brewing yeast leads to the decline of beer foam stability, especially for the unpasteurized beer. The objective of this study was to construct an industrial brewing yeast strain to secrete recombinant barley LTP1 into fermenting wort during beer fermentation for the foam stability improvement. We constructed barley LTP1 expression cassette and transformed into the host industrial yeast cells to replace partial PEP4 alleles using homologous recombination method. The expression of b-LTP1 was under control of the constitutive yeast ADH1 promoter, and the concentration of recombinant barley LTP1 secreted by recombinants reached 26.23 mg/L after incubation in YEPD medium for 120 h. The PrA activity of the recombinant strain declined compared with the host strain. The head retention of beer brewed with the recombinant industrial strain (326 ± 12 s) was improved when the host strain WZ65 (238 ± 7 s) and the constructed strain S.c-P-1 (273 ± 10 s) with partial PEP4 gene deficiency were used as control. The present study may provide reference for brewing industries and researches on beer foam stability.     

Barley for Brewing: Characteristic Changes during Malting,Brewing and Applications of its By‐Products

ABSTRACT: Barley is the basic raw material for brewing. Its chemical composition, brewing, and technological indices are highly determinative for the beer quality and the economical efficiency of the brewing process. Barley is rich in protein, carbohydrates, dietary fibers, minerals, and vitamins. The presence of nonstarch polysaccharides as mixed linkage (1‐3),(1‐4)‐β‐d ‐glucans and arabinoxylans together with the enzymes are responsible for barley modification. Malting is a complex process that involves many enzymes; important ones are α‐amylase, β‐amylase, α‐glucosidase, and limit dextrinase. During the process of malting and brewing, the by‐products left after separation of the wort are rich in protein, fibers, arabinoxylans, and β‐glucan. This review summarizes and integrates barley grain with respect to nutritional, functional, and compositional changes that take place during malting and brewing. It also explores in‐depth the several by‐products obtained after brewing and their potential for various food applications. Barley brewing by‐products offer an opportunity for cereal‐based baked and extruded products with acceptable sensory and nutritional characteristics.     

啤酒酿造过程中泡沫蛋白质的变化研究

泡沫蛋白质是啤酒泡沫的骨架,其含量和性质很大程度上决定了啤酒泡沫的质量。蛋白质Z和脂转移蛋白(LTP)是啤酒泡沫蛋白质中的关键组分,实验究考察了啤酒酿造过程中泡沫蛋白质的变化过程。结果显示:在糖化过程中,分子量为53ku及20~40ku的蛋白质被分解,蛋白质Z及LTP的含量没有太大变化;麦汁煮沸过程中蛋白质含量逐渐减少,蛋白质Z、LTP1和LTP2分别减少了11%、32%及26%;主酵过程中,蛋白质Z、LTP1和LTP2的降幅较大,分别为12%、59%和31%;后酵过程中,蛋白质Z和LTP1的含量基本不变,LTP2的含量逐渐降低,到后酵结束,LTP2的降低幅度达22%。      

Characteristics of beer produced from Korean six‐row barley with the addition of adjuncts

In this study, the effect of the addition of various amounts of adjuncts (rice, wheat, corn and potato at 10 and 20%) on the fermentation period of beer produced from Korean six‐row barley was examined. Korean six‐row barley is not suitable for brewing because of its relatively high protein/starch ratio. However, this can be offset by partially replacing the barley with adjuncts. Adjunct‐added samples were analysed and compared with the control sample made from 100% six‐row barley. All adjunct‐added samples showed changes in final beer properties (e.g. lower specific gravity, higher alcohol content, beer colour), because the initial free amino nitrogen (FAN) content was decreased and the reducing sugar content was increased. However, potato‐added samples showed a higher initial FAN level, resulting in the highest alcohol content. In addition, the colour of the potato‐added samples was darker, while the colours of the other samples were lighter. Consequently, the addition of adjuncts at a level of up to 20% had an impact on some of the quality properties of the samples in terms of fermentation period, suggesting the possibility of using Korean six‐row barley with adjuncts in brewing. Copyright © 2016 The Institute of Brewing & Distilling     

High‐Throughput Isolation of Bacteriocin‐Producing Lactic Acid Bacteria,with Potential Application in the Brewing Industry

Lactic acid bacteria (LAB) were isolated from malted cereals by means of a high‐throughput screening approach and investigated for antimicrobial activity against a range of beer‐spoiling bacteria. Putative bacteriocin‐producing strains were identified by 16S rRNA analysis and the inhibitory compounds were partially characterized. Following determination of the inhibitory spectra of the strains, an unspeciated Lactobacillus sp. UCC128, with inhibitory activity against a range of beer‐spoiling strains was subjected to further characterization. A bacteriocin was purified from this strain and analyzed by mass spectrometry to determine the weight of the protein. The result indicated that the bacteriocin was highly similar to pediocin AcH/PA‐1 from Pediococcus acidilactici. The bacteriocin‐producers identified in this study have the potential to be used in the brewing industry to enhance the microbiological stability of beer in conjunction with hurdles already in place in the brewing process.     

Part III: the influence of serial repitching of Saccharomyces pastorianus on the production dynamics of some important aroma compounds during the fermentation of barley and gluten‐free buckwheat and quinoa wort

The present paper is the last report of a comprehensive study regarding the influence of the serial repitching of Saccharomyces pastorianus TUM 34/70 on the composition of a barley, buckwheat or quinoa fermentation medium. In particular, it focuses on the production dynamics of important volatile compounds typically associated with the aroma of beer. Samples were taken every 24 h after 11 serial repitchings of a single starter culture, analysed for the particular aroma compound content by distillation followed by gas chromatography with flame ionization detection. The term ‘serial repitching factor’ is used for the first time to support the visual evaluation of the influence of serial repitching. Results showed that the levels of methanol in the quinoa wort fermentation were only slightly higher than in barley and in practical terms independent of successive fermentation. The behaviour of acetaldehyde in quinoa was similar to that in barley. However, there was a final 2‐fold lower production of some important aroma compounds compared with barley and buckwheat and for this reason quinoa cannot be recommended as a gluten‐free substitute to produce a bottom‐fermented beer. Regarding the buckwheat wort fermentation, a 2‐ to 3‐times lower final acetaldehyde content than in barley is desirable, whereas a relatively high methanol content is not desirable. Barley and buckwheat showed comparable sum concentrations and similar overall profiles of some important aroma compounds. From this perspective, buckwheat appears to be a promising substitute for barley as a brewing raw material. The overall conclusions of our comprehensive study (Parts I–III) are that buckwheat shows adequate brewing properties to substitute for barley in the commercial preparation of a bottom‐fermented gluten‐free beer‐like beverage, and yeast can be repitched at least 11 times. In contrast, quinoa in practical terms shows no substitutional potential for barley in beer; however, it has many nutritious advantages, thus the commercial preparation of a unique, bottom‐fermented gluten‐free ‘non‐beer‐like’ beverage – where the yeast could be repitched six times at most – appears feasible. Copyright © 2015 The Institute of Brewing & Distilling     

The Effect of Proteinase A on Foam‐Active Polypeptides During High and Low Gravity Fermentation

The ability of beer to produce good foam is influenced by the level of foam‐active polypeptides. Specific polypeptides with hydrophobic domains, such as Lipid Transfer Protein (LTP1), are important components of beer foam. Although, high gravity brewing is a commercially viable technique, it has the disadvantage of producing beer with less foam stability compared to lower gravity brewed counterparts. It is thought that proteinase A plays a key role in the degradation of these hydrophobic polypeptides responsible the beer foam stability. The object of this study was to compare and quantify the loss of hydrophobic polypeptides and specifically foam‐LTP1 during high gravity (20°Plato) and low gravity (12°Plato) wort fermentations and to evaluate the effect of proteinase A on these polypeptides. The losses of hydrophobic polypeptides and foam‐LTP1 were generally greater in high gravity brews. Furthermore, the results obtained suggest that proteinase A alters the hydrophobicity of these polypeptides rather than their molecular size. Approximately 20% of hydrophobic polypeptides and approximately 57% of foam‐LTP1 appeared to be proteinase A resistant. These differential losses of hydrophobic polypeptide and foam‐LTP1 could have implications for the foam stability of the finished product.     

The Impact of Malt Derived Proteins on Beer Foam Quality. Part II: The Influence of Malt Foam-positive Proteins and Non-starch Polysaccharides on Beer Foam Quality

Pilot (50 litre) and small scale (700 mL) brewing trials conducted using, similar brewing protocols with 25 different malts, indicated that differences in malt quality influenced foam stability (Rudin head retention value) by up to 24%. In addition to conventional measures of malt quality, enzyme-linked immunosorbent assays (ELISA) were used to measure the level of the putative foam-positive proteins, BSZ4 (protein Z4), BSZ7 (protein Z7), BSZ7b and lipid transfer protein 1 (LTP1). Regression analysis performed on the combined pilot and small scale data sets identified that malt BSZ4, wort β-glucan and wort viscosity, and beer protein, β-glucan and arabinoxylan were positively correlated with foam stability, while malt Kolbach index (KI), and beer FAN were negatively correlated with foam stability. Potentially foam-positive proteins such as BSZ7 and LTP1 were not correlated with foam stability. The negative correlation of BSZ4 level with KI suggested an additional role for BSZ4 in influencing protein modification. Step-wise multiple regression indicated that up to 82% of the variation in foam stability could be predicted from the malt and beer characteristics measured, demonstrating that there are a number of inter related malt derived factors that influence beer foam stability.     

Processing of bottom‐fermented gluten‐free beer‐like beverages based on buckwheat and quinoa malt with chemical and sensory characterization

Typical beer contains significant amount of gluten, and being the third most popular beverage worldwide, the commercial production of its gluten‐free form is of rising interest. This research aimed to prepare bottom‐fermented beverages from buckwheat and quinoa and to explore their physical, chemical and sensory properties. Compared with barley, the analysis of brewing attributes of buckwheat and quinoa showed a lower malt extracts, longer saccharification times, higher total protein and fermentable amino nitrogen content and higher values of the iodine test and colour. Fermentability values, the wort pH and the soluble protein content were similar for barley and buckwheat, but different for quinoa, whereas only values of viscosity and beverage pH were similar between barley and quinoa. Both beverages, especially the quinoa beverage, contained a superior level of metal cations. The fermentable carbohydrate content in the buckwheat wort was comparable to barley but lower in quinoa; however, worts derived from both pseudocereals contained predominantly glucose. The amino acid content of the buckwheat wort was similar to barley, whereas the content in the quinoa beverage was almost twice as high. The content of volatile compounds commonly associated with beer aroma was comparable between the barley and buckwheat beverage but significantly lower in the quinoa; however, the latter contained some distinctive volatile substances not found in the other beverages. The organoleptic perception of the buckwheat beverage was better than that of the quinoa, although both showed a good general acceptance. In general, buckwheat appears quite similar to barley, whereas quinoa shows many unique properties. Copyright © 2014 The Institute of Brewing & Distilling