Brewers’ spent grain (BSG) is the main by-product of the brewing industry, corresponding to ~85% of its solid residues. The attention of food technologists towards BSG is due to its content in nutraceutical compounds and its suitability to be dried, ground, and used for bakery products. This work was aimed to investigate the use of BSG as a functional ingredient in bread-making. BSGs were characterised for formulation (three mixtures of malted barley and unmalted durum (Da), soft (Ri), or emmer (Em) wheats) and origin (two cereal cultivation places). The breads enriched with two different percentages of each BSG flour and gluten were analysed to evaluate the effects of replacements on their overall quality and functional characteristics. Principal Component Analysis homogeneously grouped BSGs by type and origin and breads into three sets: the control bread, with high values of crumb development, a specific volume, a minimum and maximum height, and cohesiveness; Em breads, with high values of IDF, TPC, crispiness, porosity, fibrousness, and wheat smell; and the group of Ri and Da breads, which have high values of overall smell intensity, toasty smell, pore size, crust thickness, overall quality, a darker crumb colour, and intermediate TPC. Based on these results, Em breads had the highest concentrations of nutraceuticals but the lowest overall quality. Ri and Da breads were the best choice (intermediate phenolic and fibre contents and overall quality comparable to that of control bread). Practical applications: the transformation of breweries into biorefineries capable of turning BSG into high-value, low-perishable ingredients; the extensive use of BSGs to increase the production of food commodities; and the study of food formulations marketable with health claims.

The guarantee of the origin and quality of raw material is essential for the protection and valorization of Campana buffalo mozzarella cheese. The risk of utilization of semifinished products and stored milk in substitution for fresh milk is increasing, due to the continuous desire to reduce production costs. A proteomics approach and electrophoresis survey of retail mozzarella cheeses indicated different rates of proteolysis in the production of dairy industries. The use of fresh milk and correct cheesemaking protocol yielded only γ-caseins, which are derived from β-casein by plasmin, and para-κ-casein, which is derived from κ-casein by chymosin. The detection of abnormal hydrolysis resulting in β- and αS1-casein fragments, identified by mass spectrometry, indicates the use of stored milk or stored and pressed curd, or the reuse of unsold mozzarella cheese, to produce mozzarella. The formation of γ-caseins and other fragments during a long storage of raw materials at room or refrigeration temperature was ascribed to plasmin (endogenous milk enzyme), whereas formation of αS1-casein fragments, mainly αS1-I(6P)- and αS1-I(7P)-casein during the storage of curd was ascribed to the action of chymosin (exogenous enzyme) from rennet. Sodium dodecyl sulfate-PAGE and alkaline urea-PAGE permitted us to evaluate the freshness of the raw materials used in the manufacturing of buffalo mozzarella cheese and to reveal possible inappropriate preservation.