Environment found to explain the largest variance in physical and compositional traits in malting barley grain.

BACKGROUND: Starch is the most abundant constituent (dry weight) in the barley endosperm, followed by protein. Variability of compositional and potentially related physical traits due to genotype and environment can have important implications for the malting and brewing industry. This was the first study to assess the effects of genotype, environment, and their interaction (G × E) on endosperm texture, protein content, and starch traits corresponding to granule size, gelatinization, content, and composition, using a multi-environment variety trial in California, USA. RESULTS: Overall, environment explained the largest variance for all traits (ranging from 23.2% to 76.5%), except the endosperm texture traits wherein the G × E term explained the largest variance (45.0-86.5%). Our unique method to quantify the proportion of fine and coarse milled barley particles using laser diffraction showed a binomial distribution of endosperm texture. The number of small starch granules varied significantly (P-value < 0.05) across genotypes and environments. We observed negative correlations between total protein content and each of enthalpy (-0.70), total starch content (-0.54), and difference between offset and onset gelatinization temperature (-0.52). Furthermore, amylose to amylopectin ratio was positively correlated to volume of small starch granules (0.36). CONCLUSION: Our findings indicate that environment played a larger role in influencing the majority of starch-related physical and compositional traits. In contrast, variance in endosperm texture was largely explained by G × E. Maltsters would benefit from accounting for environmental contributions in addition to solely genotype when making sourcing decisions, especially with regards to total protein, total starch, enthalpy, and difference between offset and onset gelatinization temperature. © 2024 Society of Chemical Industry.

Proteomics and Metabolomics Reveal that an Abundant α-Glucosidase Drives Sorghum Fermentability for Beer Brewing.

Sorghum (Sorghum bicolor), a grass native to Africa, is a popular alternative to barley for brewing beer. The importance of sorghum to beer brewing is increasing because it is a naturally gluten-free cereal, and climate change is expected to cause a reduction in the production of barley over the coming decades. However, there are challenges associated with the use of sorghum instead of barley in beer brewing. Here, we used proteomics and metabolomics to gain insights into the sorghum brewing process to advise processes for efficient beer production from sorghum. We found that during malting, sorghum synthesizes the amylases and proteases necessary for brewing. Proteomics revealed that mashing with sorghum malt required higher temperatures than barley malt for efficient protein solubilization. Both α- and ß-amylase were considerably less abundant in sorghum wort than in barley wort, correlating with lower maltose concentrations in sorghum wort. However, metabolomics revealed higher glucose concentrations in sorghum wort than in barley wort, consistent with the presence of an abundant α-glucosidase detected by proteomics in sorghum malt. Our results indicate that sorghum can be a viable grain for industrial fermented beverage production, but that its use requires careful process optimization for efficient production of fermentable wort and high-quality beer.

Large-scale screening of yeast strains that can utilize proline.

Proline contributes to the taste and flavor of foods. The yeast Saccharomyces cerevisiae poorly assimilates proline during fermentation processes, resulting in the accumulation of proline in fermentative products. We performed here a screening of in total 1138 yeasts to obtain strains that better utilize proline. Our results suggest that proline utilization occurs in the genera of Zygoascus, Galactomyces, and Magnusiomyces.

Humulus lupulus and microbes: Exploring biotic causes for hop creep.

BACKGROUND: Hop creep continues to present an unresolved issue for the brewing industry, specifically stemming from those hops added to beer during fermentation. Hops have been found to contain four dextrin-degrading enzymes: alpha amylase, beta amylase, limit dextrinase, and an amyloglucosidase. One recent hypothesis predicts that these dextrin-degrading enzymes could originate from microbes rather than the hop plant itself. SCOPE AND APPROACH: This review begins by describing how hops are processed and used in the brewing industry. It will then discuss hop creep's origins with a new beer style, antimicrobial factors from hops and resistance mechanisms that bacteria use to counter them, and finally microbial communities that inhabit hops, focusing on whether they can produce the starch degrading enzymes which drive hop creep. After initial identification, microbes with possible links to hop creep were then run through several databases to search the genomes (if available) and for those specific enzymes. KEY FINDINGS AND CONCLUSIONS: Several bacteria and fungi contain alpha amylase as well as unspecified glycosyl hydrolases, but only one contains beta amylase. Finally, this paper closes with a short summary of how abundant these organisms typically are in other flowers.

The Cdc25/Ras/cAMP-dependent protein kinase A signaling pathway regulates proline utilization in wine yeast Saccharomyces cerevisiae under a wine fermentation model.

Proline is a predominant amino acid in grape must, but it is poorly utilized by the yeast Saccharomyces cerevisiae in wine-making processes. This sometimes leads to a nitrogen deficiency during fermentation and proline accumulation in wine. In this study, we clarified that a glucose response is involved in an inhibitory mechanism of proline utilization in yeast. Our genetic screen showed that strains with a loss-of-function mutation on the CDC25 gene can utilize proline even under fermentation conditions. Cdc25 is a regulator of the glucose response consisting of the Ras/cAMP-dependent protein kinase A (PKA) pathway. Moreover, we found that activation of the Ras/PKA pathway is necessary for the inhibitory mechanism of proline utilization. The present data revealed that crosstalk exists between the carbon and proline metabolisms. Our study could hold promise for the development of wine yeast strains that can efficiently assimilate proline during the fermentation processes.

The intrinsic and regulated proteomes of barley seeds in response to fungal infection.

Barley is an important cereal grain used for beer brewing, animal feed, and human food consumption. Fungal disease can impact barley production, as it causes substantial yield loss and lowers seed quality. We used sequential window acquisition of all theoretical ions mass spectrometry (SWATH-MS) to measure and quantify the relative abundance of proteins within seeds of different barley varieties under various fungal pathogen burdens (ProteomeXchange Datasets PXD011303 and PXD014093). Fungal burden in the leaves and stems of barley resulted in changes to the seed proteome. However, these changes were minimal and showed substantial variation among barley samples infected with different pathogens. The limited effect of intrinsic disease resistance on the seed proteome is consistent with the main mediators of disease resistance being present in the leaves and stems of the plant. The seeds of barley varieties accredited for use as malt had higher levels of proteins associated with starch synthesis and beer quality. The proteomic workflows developed and implemented here have potential application in quality control, breeding and processing of barley, and other agricultural products.

Curcumin-based photosensitization inactivates Aspergillus flavus and reduces aflatoxin B1 in maize kernels.

Different methods have been applied in controlling contamination of foods and feeds by the carcinogenic fungal toxin, aflatoxin, but nevertheless the problem remains pervasive in developing countries. Curcumin is a natural polyphenolic compound from the spice turmeric (Curcuma longa L.) that has been identified as an efficient photosensitiser for inactivation of Aspergillus flavus conidia. Curcumin mediated photoinactivation of A. flavus has revealed the potential of this technology to be an effective method for reducing population density of the aflatoxin-producing fungus in foods. This study demonstrates the influence of pH and temperature on efficiency of photoinactivation of the fungus and how treating spore-contaminated maize kernels affects aflatoxin production. The results show the efficiency of curcumin mediated photoinactivation of fungal conidia and hyphae were not affected by temperatures between 15 and 35 °C or pH range of 1.5-9.0. The production of aflatoxin B1 was significantly lower (p < 0.05), with an average of 82.4 µg/kg as compared to up to 305.9 µg/kg observed in untreated maize kept under similar conditions. The results of this study indicate that curcumin mediated photosensitization can potentially be applied under simple environmental conditions to achieve significant reduction of post-harvest contamination of aflatoxin B1 in maize.

Process Proteomics of Beer Reveals a Dynamic Proteome with Extensive Modifications.

Modern beer production is a complex industrial process. However, some of its biochemical details remain unclear. Using mass spectrometry proteomics, we have performed a global untargeted analysis of the proteins present across time during nanoscale beer production. Samples included sweet wort produced by a high temperature infusion mash, hopped wort, and bright beer. This analysis identified over 200 unique proteins from barley and yeast, emphasizing the complexity of the process and product. We then used data independent SWATH-MS to quantitatively compare the relative abundance of these proteins throughout the process. This identified large and significant changes in the proteome at each process step. These changes described enrichment of proteins by their biophysical properties, and identified the appearance of dominant yeast proteins during fermentation. Altered levels of malt modification also quantitatively changed the proteomes throughout the process. Detailed inspection of the proteomic data revealed that many proteins were modified by protease digestion, glycation, or oxidation during the processing steps. This work demonstrates the opportunities offered by modern mass spectrometry proteomics in understanding the ancient process of beer production.

Sequencing of Australian wild rice genomes reveals ancestral relationships with domesticated rice.

The related A genome species of the Oryza genus are the effective gene pool for rice. Here, we report draft genomes for two Australian wild A genome taxa: O. rufipogon-like population, referred to as Taxon A, and O. meridionalis-like population, referred to as Taxon B. These two taxa were sequenced and assembled by integration of short- and long-read next-generation sequencing (NGS) data to create a genomic platform for a wider rice gene pool. Here, we report that, despite the distinct chloroplast genome, the nuclear genome of the Australian Taxon A has a sequence that is much closer to that of domesticated rice (O. sativa) than to the other Australian wild populations. Analysis of 4643 genes in the A genome clade showed that the Australian annual, O. meridionalis, and related perennial taxa have the most divergent (around 3 million years) genome sequences relative to domesticated rice. A test for admixture showed possible introgression into the Australian Taxon A (diverged around 1.6 million years ago) especially from the wild indica/O. nivara clade in Asia. These results demonstrate that northern Australia may be the centre of diversity of the A genome Oryza and suggest the possibility that this might also be the centre of origin of this group and represent an important resource for rice improvement.

Applications of single kernel conventional and hyperspectral imaging near infrared spectroscopy in cereals.

Single kernel (SK) near infrared (NIR) reflectance and transmittance technologies have been developed during the last two decades for a range of cereal grain physical quality and chemical traits as well as detecting and predicting levels of toxins produced by fungi. Challenges during the development of single kernel near infrared (SK-NIR) spectroscopy applications are modifications of existing NIR technology to present single kernels for scanning as well as modifying reference methods for the trait of interest. Numerous applications have been developed, and cover almost all cereals although most have been for key traits including moisture, protein, starch and oil in the globally important food grains, i.e. maize, wheat, rice and barley. An additional benefit in developing SK-NIR applications has been to demonstrate the value in sorting grain infected with a fungus or mycotoxins such as deoxynivalenol, fumonisins and aflatoxins. However, there is still a need to develop cost-effective technologies for high-speed sorting which can be used for small grain samples such as those from breeding programmes or commercial sorting; capable of sorting tonnes per hour. Development of SK-NIR technologies also includes standardisation of SK reference methods to analyse single kernels. For protein content, the use of the Dumas method would require minimal standardisation; for starch or oil content, considerable development would be required. SK-NIR, including the use of hyperspectral imaging, will improve our understanding of grain quality and the inherent variation in the range of a trait. In the area of food safety, this technology will benefit farmers, industry and consumers if it enables contaminated grain to be removed from the human food chain.

Non-invasive exploration of malting barley (Hordeum vulgare) in vitro germination and varietal effects using short wave infrared spectral imaging and ANOVA simultaneous component analysis.

ANOVA-simultaneous component analysis (ASCA) was applied to short-wave infrared spectral fingerprints of 5 malting barley varieties collected using a hyperspectral imaging system to determine the effect of germination, the influence of time and the influence of barley by means of a full factorial experimental design. ASCA indicated that there was a significant (p < 0.0001) effect of the germination status, the germination time and interaction on the spectral data for all varieties. The biochemical and physiological modification of the samples were characterised by visualisation of the longitudinal scores obtained from simultaneous component analysis for the germination time factor. This resulted in the visualisation and explanation of biochemical change over the course of barley germination as a factor of time. The relevant loadings indicated a significant change to the proteome, lipid and starch structure as driven by the uptake of water over time. The ASCA model were extrapolated to include the effect of barley variety to the already mentioned germination status and germination time factors, resulting once again in all the effects being significant (p < 0.0001). Here it was shown that all the barley varieties are significantly different from one another pre- and post-modification, based on the molecular vibrations observed in the short wave-infrared (SWIR) spectra, suggesting that the detection of biotic stress factors, such as pre-harvest germination, also differ for each variety, by indicating that the germination profile of each barley variety varies as a function of germination time. Thus, also the malting performance, germinative energy and chemical profile of each barley variety tested will vary before, during and after imbibition and germination - indicating the importance of malting commercial barley malt true to variety. These results indicate that (SWIR) spectral imaging instrumentation can possibly be used to monitor controlled germination of barley grain. Due to the shown ability of SWIR spectral imaging to detect small biochemical changes over time of barley grain during germination.

Analysis of mousy off-flavor compound 2-Acetyl-tetrahydropyridine using Liquid Chromatography Mass Spectrometry with Electrospray Ionization in sour beer.

Mousy off-flavor describes N-heterocycles compounds related to spoilage in the brewing industry. It has also been identified in sour beers through sensory analysis. Therefore, preventing spoilage N-heterocycles development is essential to preserve end-products and obviate economic losses. To this day, no methods or protocols have been reported to identifying mousy off-flavor compounds in a beer matrix. The main objective of this work was to develop a standardized quantification method for 2-acetyl-3,4,5,6-tetrahydropyridine (ATHP) in beer matrix, by Liquid Chromatography Mass Spectrometry with Electrospray Ionization (LC-MS-ESI). Extraction of ATHP in the samples was performed using QuEChERS (quick, easy, cheap, effective, rugged, and safe) technique. Over a dozen different potentially mousy cask-aged sour beers including other spontaneously fermented beverages were provided, based on sensory analysis, to determine the variation in ATHP levels. Results indicated ATHP was found in all the samples, ranging from 1.64 ± 0.06 to 57.96 ± 2.15 µg L-1. Herein, we described our detection method of mousy-off flavor compounds which enables future research to mitigate the occurrence of such defects in fermented beverages matrix.•ATHP content in samples varied from 1.64 ± 0.06 to 57.96 ± 2.15 µg L-1.•The recovery range of ATHP using LC-MS-ESI varied from 71% to 97%.•Basified QuEChERS salting-out procedure is applicable for ATHP extraction from beer and other fermented beverages matrices.

A Review of N-Heterocycles: Mousy Off-Flavor in Sour Beer.

Beer has over 600 flavor compounds and creates a positive tasting experience with acceptable sensory properties, which are essential for the best consumer experience. Spontaneous and mixed-culture fermentation beers, generally classified as sour beers, are gaining popularity compared to typical lager or ale styles, which have dominated in the USA for the last few decades. Unique and acceptable flavor compounds characterize sour beers, but some unfavorable aspects appear in conjunction. One such unfavorable flavor is called "mousy". This description is usually labeled as an unpleasant odor, identifying spoilage of fermented food and beverages. It is related as having the odor of mouse urine, cereal, corn tortilla chips, or freshly baked sour bread. The main compounds responsible for it are N-heterocyclic compounds: 2-acetyltetrahydropyridine, 2-acetyl-1-pyrroline, and 2-ethyltetrahydropyridine. The most common beverages associated with mousy off-flavor are identified in wines, sour beers, other grain-based beverages, and kombucha, which may contain heterofermentative lactic acid bacteria, acetic acid bacteria, and/or yeast/fungus cultures. In particular, the fungal species Brettanomyces bruxellensis are associated with mousy-off flavor occurrence in fermented beverages matrices. However, many factors for N-heterocycle formation are not well-understood. Currently, the research and development of mixed-cultured beer and non/low alcohol beverages (NABLAB) has increased to obtain the highest quality, sensory, functionality, and most notably safety standards, and also to meet consumers' demand for a balanced sourness in these beverages. This paper introduces mousy off-flavor expression in beers and beverages, which occurs in spontaneous or mixed-culture fermentations, with a focus on sour beers due to common inconsistency aspects in fermentation. We discuss and suggest possible pathways of mousy off-flavor development in the beer matrix, which also apply to other fermented beverages, including non/low alcohol drinks, e.g., kombucha and low/nonalcohol beers. Some precautions and modifications may prevent the occurrence of these off-flavor compounds in the beverage matrix: improving raw material quality, adjusting brewing processes, and using specific strains of yeast and bacteria that are less likely to produce the off-flavor. Conceivably, it is clear that spontaneous and mixed culture fermentation is gaining popularity in industrial, craft, and home brewing. The review discusses important elements to identify and understand metabolic pathways, following the prevention of spoilage targeted to off-flavor compounds development in beers and NABLABs.

Variation in quality of grains used in malting and brewing.

Cereal grains have been domesticated largely from food grains to feed and malting grains. Barley (Hordeum vulgare L.) remains unparalleled in its success as a primary brewing grain. However, there is renewed interest in "alternative" grains for brewing (and distilling) due to attention being placed on flavor, quality, and health (i.e., gluten issues) aspects that they may offer. This review covers basic and general information on "alternative grains" for malting and brewing, as well as an in-depth look at several major biochemical aspects of these grains including starch, protein, polyphenols, and lipids. These traits are described in terms of their effects on processing and flavor, as well as the prospects for improvement through breeding. These aspects have been studied extensively in barley, but little is known about the functional properties in other crops for malting and brewing. In addition, the complex nature of malting and brewing produces a large number of brewing targets but requires extensive processing, laboratory analysis, and accompanying sensory analysis. However, if a better understanding of the potential of alternative crops that can be used in malting and brewing is needed, then significantly more research is required.

Isolation of Yeast Strains with Higher Proline Uptake and Their Applications to Beer Fermentation.

Although proline is the most or second most abundant amino acid in wort and grape must, it is not fully consumed by the yeast Saccharomyces cerevisiae during alcoholic fermentation, unlike other amino acids. Our previous studies showed that arginine, the third most abundant amino acid in wort, inhibits the utilization of proline in most strains of S. cerevisiae. Furthermore, we found that some non-Saccharomyces yeasts utilized proline in a specific artificial medium with arginine and proline as the only nitrogen source, but these yeasts were not suitable for beer fermentation due to their low alcohol productivity. For yeasts to be useful for brewing, they need to utilize proline and produce alcohol during fermentation. In this study, 11 S. cerevisiae strains and 10 non-Saccharomyces yeast strains in the Phaff Yeast Culture Collection were identified that utilize proline effectively. Notably, two of these S. cerevisiae strains, UCDFST 40-144 and 68-44, utilize proline and produce sufficient alcohol in the beer fermentation model used. These strains have the potential to create distinctive beer products that are specifically alcoholic but with a reduction in proline in the finished beer.

Effect of processing on the solubility and molecular size of oat ß-glucan and consequences for starch digestibility of oat-fortified noodles.

White wheat salted noodles containing oats have a slower digestion rate those without oats, with potential health benefits. Oat ß-glucan may play an important role in this. Effects of sheeting and shearing during noodle-making and subsequent cooking on ß-glucan concentration, solubility, molecular size and starch digestibility were investigated. The levels of ß-glucan were reduced by 16% after cooking, due to the loss of ß-glucan into the cooking water. Both the noodle-making process and cooking increased the solubility of ß-glucan but did not change its average molecular size. Digestion profiles show that ß-glucan in wholemeal oat flour did not change starch digestion rates compared with isolated starch, but reduced the starch digestion rate of oat-fortified wheat noodles compared to the control (wheat noodles). Confocal laser scanning microscopy suggests that interaction between ß-glucan and protein contributes to the starch-protein matrix and changes noodle microstructure, and thus alters their digestibility.

A review of triticale uses and the effect of growth environment on grain quality.

Triticale (× Triticosecale sp. Wittmack ex A. Camus 1927) is an anthropogenic cereal designed to incorporate the functionality and high yield of wheat (Triticum spp. Linnaeus 1753) and durability of rye (Secale cereale Linnaeus 1753). The potential of triticale has remained largely unrealised, and in the 135 years since A. Stephen Wilson first crossed wheat and rye, triticale has mostly been used as animal feed. Growing demand for food resources has led to an increased interest in triticale development. Efforts to breed cultivars appropriate for baking have met with difficulty, although relatively new approaches to triticale end-use propose greater applicability for human consumption. Further, environmental awareness has generated interest in the use of triticale within biofuel production. We review environmental and genetic effects on triticale yield with a view towards increased demand on a hardy and useful cereal crop. We find triticale could satisfy many of the hopes originally placed upon it, and may be useful in foodstuffs and fuel, but only when growth environment is carefully considered.

Fungal Biovalorization of a Brewing Industry Byproduct, Brewer's Spent Grain: A Review.

The beer industry is a major producer of solid waste globally, primarily in the form of brewer's spent grain (BSG), which due to its low value has historically been diverted to livestock as feed or to landfills. However, its high moisture content and chemical composition positions BSG as an ideal candidate for further processing with microbial fermentation. Recent research has focused on filamentous fungi and the ability of some species therein to degrade the predominant recalcitrant cellulolignin components of BSG to produce valuable compounds. Many species have been investigated to biovalorize this waste stream, including those in the genuses Aspergillus, Penicillium, Rhyzopus, and Trichoderma, which have been used to produce a wide array of highly valuable enzymes and other functional compounds, and to increase the nutritional value of BSG as an animal feed. This review of recent developments in the application of filamentous fungi for the valorization of BSG discusses the biochemical makeup of BSG, the biological mechanisms underlying fungi's primacy to this application, and the current applications of fungi in this realm.

Protein-starch matrix plays a key role in enzymic digestion of high-amylose wheat noodle.

Wheat flour, consisting of a complex matrix of starch and protein, is used as a representative model of whole food here to investigate the binary interaction in relation to amylose level and hydrothermal treatment in noodles as a food exemplar. Noodle made of high-amylose wheat (HAW) flour showed an eight-fold higher resistant starch content, compared to the wild type. Protein removal under simulated intestinal digestion conditions resulted in higher starch digestion rate coefficients in raw and cooked flours. In cooked flours, the substrate becomes similarly accessible to digestive enzymes regardless of protein removal. The results indicate that the increased protein content in native HAW flour and thermal stability of starch in HAW noodles lead to higher food integrity and consequently enhance the resistance against α-amylase digestion. Overall, the study suggests that a diversity of starch-protein interactions in wheat-based food products underlies the nutritional value of natural whole foods.

Late-Maturity Alpha-Amylase in Wheat (Triticum aestivum) and Its Impact on Fresh White Sauce Qualities.

When wheat experiences a cold-temperature 'shock' during the late stage of grain filling, it triggers the abnormal synthesis of late-maturity α-amylase (LMA). This increases the enzyme content in affected grain, which can lead to a drastic reduction in falling number (FN). By commercial standards, a low FN is taken as an indication of inferior quality, deemed unsuitable for end-product usage. Hence, LMA-affected grains are either rejected or downgraded to feed grade at the grain receiving point. However, previous studies have found no substantial correlation between low FN-LMA and bread quality. The present study extends previous investigations to semi-solid food, evaluating the physical quality of fresh white sauce processed from LMA-affected flour. Results show that high-LMA flours had low FNs and exhibited poor pasting characteristics. However, gelation occurred in the presence of other components during fresh white sauce processing. This demonstrates that LMA-affected flours may have new applications in low-viscosity products.