In this study, 40% of unmalted gluten free (GF) grains (sorghum, millet, buckwheat, quinoa and amaranth) was used in brewing process, in gelatinized and ungelatinized form, in order to produce GF beer and to extend current knowledge about their suitability as brewing adjuncts. Partial replacement of barley malt with GF grains led to a significant decrease of extract (°P) and alcohol (%v/v) content compared to control beer (p < 0.05), except for quinoa beer (QB). Results from Principal Component Analysis (PCA) highlighted a satisfactory classification of experimental beers according to the two different forms of GF grains (gelatinized and ungelatinized). However, beers brewed with ungelatinized grains (mainly sorghum and quinoa) showed acceptable technological and sensory properties, thus suggesting that the pre-gelatinization step could be bypassed with a view to more environmentally and economically sustainable time-saving process. In addition, all beer samples showed a gluten content higher than 20 ppm.

There is a growing demand in reusing agro-industrial waste to enrich food proteins and develop new protein-related products. Brewer's spent grain (BSG), the byproducts generated from the processing of beer-brewing industry, is rich in abundant protein (26 to 30%) with good physicochemical properties and nutritional benefits. BSG is mainly composed of four proteins including hordein, gluten, globulin, and albumin. The methods for extracting protein from BSG mainly include alkali extraction, ultrasonic-assisted extraction, and organic solvent extraction, among others. However, little researches have described the functional properties of Brewer's spent grain protein (BSGP) and how it can be improved by enzymatic modification. Additionally, BSG protein hydrolysates (BSGPHs) have good bioactivities, mainly including antioxidant, antiinflammatory, and angiotensin-I-converting enzyme inhibitory activities, among others. Based on the above situation, this review provides a comprehensive overview about the isolation, physicochemical features, application of the BSGP. In addition, the functional properties and biological activities of BSGPHs are also emphasized. The purpose of this review is to provide an up-to-date summary of BSGP research, and to broaden the market potential of this emerging protein in food industry. Besides, this review provides a reference to study the use of BSGP or hydrolysates for a variety of purposes in-depth, including pharmaceutical, food, and industrial applications.

Brewer’s spent grain (BSG) is the major side-stream from beer production but remains highly underutilized. While the direct use of BSG as a food ingredient is limited due to subpar techno- functionality, the vast amounts and fairly high protein content of up to 30% makes it a high potential source for production of protein-based ingredient by valorization through e.g. enzymatic hydrolysis. However, little attention has been put towards the protein-level composition of BSG, which is essential for developing hydrolysis strategies for improving functionality in a targeted manner. Here, we present an in-depth characterization of the BSG proteome and investigate dynamic proteome changes from malting and mashing in the initial phases of beer production. We show dynamic and selective changes in the proteome across the different process steps, where 29% of reproducibly identified proteins display differential abundance. BSG represents a significantly higher proportion of intracellular protein compared to both barley and malt and has a nutritionally favorable amino acid composition. The major constituent of the BSG proteome is B3-Hordein, constituting more than 30% of the BSG protein. Moreover, we find that a large proportion (> 45%) of the BSG protein is associated with potential food safety concerns, being classified as potential allergens and antinutritional factors. Our analysis emphasizes the need for downstream processing of BSG to produce safe and functional food ingredients, while also providing protein-level insights for development of targeted hydrolysis strategies to achieve this. Highlights An optimized sample preparation for proteomics analysis has been developed 29% of barley proteins are differentially abundant across malting and mashing B3-Hordein is the major protein in BSG with an abundance over 30% BSG contains a high content of potential allergenic and antinutritional proteins A protein-level basis for targeted downstream processing of BSG is presented

Grain proteins in cereal crops play a crucial role in determining both the nutritional value and end-use quality of food products. This systematic review comprehensively examines the biosynthetic pathways, regulatory mechanisms, and functional impacts of storage proteins in rice (<i>Oryza sativa</i>) and barley (<i>Hordeum vulgare</i>), two of the world's most important staple crops. Rice and barley are significant sources of bioactive proteins, which are fundamental to their nutritional value and health promoting properties. The major storage proteins, including glutelins, prolamins (with hordeins being the major type in barley), and globulins, are synthesized under the regulation of key transcription factors like <i>RISBZ</i> and <i>RPBF</i> in rice and <i>BLZ1/2</i> in barley. These proteins provide essential amino acids and are a source of bioactive peptides with demonstrated anti-hypertensive, immunomodulatory, and cholesterol-lowering activities. Furthermore, the review considers the influence of genetic and environmental factors on protein profiles. The health implications of rice and barley proteins are discussed, underscoring their potential in functional foods and nutraceuticals. Future perspectives highlight the promise of metabolic engineering and precision breeding for the biofortification and nutritional enhancement of these vital cereals.

Recent reports have highlighted that beer labelled "gluten-free", crafted with enzymatic treatments to remove gluten, may contain polypeptides that could be immunotoxic to individuals with coeliac disease. As strict adherence to a gluten-free diet is the only way to manage this condition, accurate labelling is crucial to those with coeliac disease. This paper aims to discuss the presence, levels and immunogenicity of gluten peptides found in gluten-reduced barley beers. While advances have been made in the detection and quantification of gluten peptides in beer, there are still challenges to the interpretation of gluten measurements as well as to assess whether peptides are immunotoxic in vivo. To make progress, future efforts should involve a combination of in vivo toxicity assessment of the degraded proteins, development of standardised gluten-free production strategies to minimise variability in gluten fragment presence, guidance on how to control the outcome as well as to develop appropriate reference materials and calibrators.

Celiac disease (CD), a chronic enteropathy of the small intestine caused by ingestion of gluten, is one of the most prevalent food hypersensitivities worldwide. The essential treatment is a strict lifelong gluten-free diet based on the avoidance of gluten-containing products from wheat, rye, barley and, in rare cases, oats. Products made from naturally gluten-free raw materials often have inferior nutritional, textural and sensory properties compared to the corresponding gluten-containing products. Therefore, the incorporation of wheat, rye and barley flours after efficient removal of the harmful component gluten into gluten-free products would be beneficial. Gluten modification resulting in decreased CD-immunoreactivity may be achieved via the formation of crosslinks using microbial transglutaminase. To effectively eliminate CD-immunoreactivity, plant, fungal, bacterial, animal or engineered peptidases are capable of degrading gluten proteins and peptides into harmless fragments. The application of peptidases from germinated cereal grains, fungal peptidases and/or lactic acid bacteria during food processing yielded high-quality sourdough wheat breads, pasta, wheat starch and bran, rye products and beer, all with gluten contents below the Codex Alimentarius threshold of 20mg/kg for gluten-free products. As with all gluten-free products, the legislative compliance of such treated materials needs to be monitored closely. Provided that all safety requirements are met, gluten-containing raw materials treated in an adequate way to remove CD-active gluten fragments may be used together with naturally gluten-free ingredients to create an extended choice of high-quality gluten-free products.

Gluten-free regulations require analytical systems that can reliably measure gluten in wheat, rye, and barley, and, in some jurisdictions, in oats across diverse food ingredients and processed foods. Examples include baked goods, fermented products such as beer, and extruded snack foods, where processing can alter gluten extractability and antibody recognition. This review examines the scientific, analytical, and regulatory factors that determine whether gluten quantification can support robust enforcement of the 20 mg gluten/kg threshold used in most jurisdictions, with a focus on method performance criteria (MPCs). The molecular complexity of gluten and the structural changes induced by baking, fermentation, and other processing steps influence extractability and epitope availability. These effects contribute to variable results across enzyme-linked immunosorbent assays that use different antibodies, extraction chemistries, and calibration standards. The widespread use of PWG gliadin (gliadin reference material) as a calibrant supports harmonized calibration for wheat-based analyses. Still, it can introduce bias because it does not reflect the gluten composition in processed or mixed-cereal foods. Improved incurred reference materials offer advantages but are not yet widely accessible. Proficiency testing data from DLA and FAPAS show that interlaboratory agreement is strongest for unprocessed matrices and decreases in baked, fat-rich, or fermented foods. These findings highlight the need for harmonized validation and for statistical criteria that define acceptable assay performance around the regulatory threshold. Advanced techniques such as liquid chromatography-tandem mass spectrometry provide detailed peptide-level information and can support confirmation in matrices where immunoassays lose sensitivity. A coordinated, performance-based framework aligned with Codex guidance would enable analytical methods to be evaluated against common criteria for recovery, precision, and detection capability. Codex alignment also introduces method dependency, as the current Type I Codex approach for gluten relies on the R5 Méndez ELISA as the defining method. Because Type I methods are empirical defining methods, results are comparable to the definition established by that method rather than necessarily being traceable to an independent reference value for gluten. The use of a single Type I method can therefore limit comparability when laboratories use assays based on other antibodies or extraction and calibration systems, particularly in processed matrices. Integrating such MPCs into regulatory practice, proficiency testing, and private certification systems would improve the comparability of gluten testing results and improve the reliability of gluten-free labeling in international markets.

Background and Aims: Celiac disease is a permanent intolerance to gluten prolamins from wheat, barley, rye and, in some patients, oats. Partially digested gluten peptides produced in the digestive tract cause inflammation of the small intestine. High throughput, immune-based assays using monoclonal antibodies specific for these immunotoxic peptides would facilitate their detection in food and enable monitoring of their enzymatic detoxification. Two monoclonal antibodies, G12 and A1, were developed against a highly immunotoxic 33-mer peptide. The potential of each antibody for quantifying food

Gluten-free beer is beer made from ingredients that do not contain gluten, such as millet, rice, sorghum, buckwheat or corn (maize). People who have gluten Gluten-free beer is beer made from ingredients that do not contain gluten, such as millet, rice, sorghum, buckwheat or corn (maize). People who have gluten intolerance (including celiac disease and dermatitis herpetiformis sufferers) have a reaction to certain proteins in the grains commonly used to make beer, barley and wheat. The hordein found in barley and the gliadin found in wheat are types o Gluten-free beer is beer made from ingredients that do not contain gluten, such as millet, rice, sorghum, buckwheat or corn (maize). People who have gluten intolerance (including celiac disease and dermatitis herpetiformis sufferers) have a reaction to certain proteins in the grains commonly used to make beer, barley and wheat. The hordein found in barley and the gliadin found in wheat are types of gluten that can trigger symptoms in sufferers of these diseases. Gluten-free beer is part of a gluten-free diet. Beers brewed mainly from cereals such as millet, rice, sorghum, buckwheat and corn (maize), which do not contain gluten, do not trigger an autoimmune response in celiacs. Some brewers brew with barley or rye, and reduce the level of gluten to below 20 ppm. This may be achieved by using enzymes such as Clarex, which break down gluten proteins in beer brewed with barley, as well as helping to filter the brew. In most countries this technically classifies them as gluten-free beers, but in the United States, they are classified as gluten-reduced beers. These brewers believe they are safe to drink. The brewers argue that the proteins from barley are converted into non-harmful amino acids. Statements from brewers show that their scientists feel confident that their product is non-harmful to those who are gluten intolerant. Some celiacs report problems drinking these beers. However, there is some concern and evidence that the claim is not true.(for example: Sheehan, Evans & Skerritt, 2001). Brewers who produce low gluten beers are required to test every batch for gluten, and record gluten levels in "parts per million" ('ppm'). Although the barley hordein in such tests may not be detected, smaller pieces of these proteins, known as peptides, may remain and be toxic for celiacs. Those involved in gluten-free brewing, and others representing celiacs or those with other conditions that require a gluten-free diet, tend to be concerned that beer brewed using wheat or barley are not appropriate for those with celiacs or dermatitis herpetiformis, although the carefully controlled gluten levels of particular malt brews of England and Finland may be low enough to be…