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This work deals with the study on the protein extractability of biscuits incurring different percentages of roasted peanut flour. The presence of two different flours influenced the rate of protein aggregation and protein extractability, according to the percentage of roasted peanut flour added to the formulation and assessing these features by testing the use of two buffers. Results showed that gluten network arrangement of biscuits was influenced by the flours mixture besides the baking, with possible different protein organizations. Protein extractability was affected, underlining a higher content of protein aggregates at high molecular weight especially with the addition of 20% of peanut flour, characterized by hydrophobic and reducible covalent bonds, as suggested by the higher extractability obtained with the buffer with chaotropic and reducing agents. These results suggested a possible induced supramolecular protein organization in these products, which could affect the immunoreactivity of the main allergens occurred in the formulation., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. The following are the supplementary data related to this article. Supplementary data to this article can be found online at https://doi.org/10.1016/j.foodchem.2024.140568., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Honey is a valuable reservoir of lactic acid bacteria (LAB) and, particularly, of fructophilic LAB (FLAB), a relatively novel subgroup of LAB whose functional potential for human and food application has yet to be explored. In this study, FLAB and LAB strains have been isolated from honeys of different floral origins and selected for their broad antimicrobial activity against typical foodborne pathogenic bacteria and spoilage filamentous fungi. The best candidates, two strains belonging to the species Lactiplantibacillus plantarum and Fructobacillus fructosus, were submitted to partial characterisation of their cell free supernatants (CFS) in order to identify the secreted metabolites with antimicrobial activity. Besides, these strains were examined to assess some major functional features, including in vitro tolerance to the oro-gastrointestinal conditions, potential cytotoxicity against HT-29 cells, adhesion to human enterocyte-like cells and capability to stimulate macrophages. Moreover, when the tested strains were applied on table grapes artificially contaminated with pathogenic bacteria or filamentous fungi, they showed a good ability to antagonise the growth of undesired microbes, as well as to survive on the fruit surface at a concentration that is recommended to develop a probiotic effect. In conclusion, both LAB and FLAB honey-isolated strains characterised in this work exhibit functional properties that validate their potential use as biocontrol agents and for the design of novel functional foods. We reported antimicrobial activity, cytotoxic evaluation, probiotic properties and direct food application of a F. fructosus strain, improving the knowledge of this species, in particular, and on FLAB, more generally., (© 2022. The Author(s).)

The incorporation of 5 and 10% freeze-dried grape pomace powder (GPP) in fresh tagliatelle pasta preparation was evaluated for its effect on chemical composition, gluten protein structure, and sensory properties. The addition of the freeze-dried GPP led to a significant increase ( p < 0.05) in polyphenol content in the raw and cooked fortified pasta samples with respect to 100% semolina pasta, although the phenolic content decreased after the cooking process. The opposite phenomenon was observed with the antioxidant activity, which increased significantly ( p < 0.05) when switching from raw to cooked pasta samples fortified with GPP. The formation of a proper gluten structure was found in the fortified raw pasta, even if a change in the protein arrangement was shown in the fortified cooked samples, confirmed by a significant reduction ( p < 0.05) in both the unextractable polymeric protein percentage (% UPP) and disulfide bond (S-S) formation. These results suggest a possible interaction between the protein sulfhydryl groups (-Cys) and polyphenols of grape pomace during cooking through non-disulfide covalent bonds, which was confirmed by the significant ( p < 0.05) decrease in the -SH groups when comparing 100% semolina pasta with fortified pasta sample. Finally, a sensory analysis showed that the highest significant score ( p < 0.05) was achieved by the 5% GP-fresh pasta sample.

Coppa Piacentina is considered a peculiar dry cured salami, since it is manufactured by the entire neck muscles stuffed and matured in natural casings, the same as dry cured ham and fermented dry cured sausages. In this work the proteolysis of external and internal portions was investigated by a proteomic approach and by amino acids analysis. Samples of "Coppa Piacentina" were analyzed at 0 days and after 5 and 8 months of ripening through mono- and two-dimensional gel electrophoresis. Image analysis of 2D electrophoretic maps indicated a more intense enzyme activity on the external part, mainly due to endogenous enzymes. They favored, respectively, myofibrillar or sarcoplasmic proteins at 5 or 8 months of ripening. Free amino acids determination proved that lysine and glutamic acid were the most represented ones, followed by a free amino acids sequence like that of dry cured ham. The peculiarities of "Coppa Piacentina" were characterized by a slow proteolysis, due to sacking and binding of the whole cut of the pork neck., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

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 molecular weight distribution of protein aggregates from raw meat and cooked pork products was assessed by size exclusion-high performance liquid chromatography (SE-HPLC). Electrophoretic analysis under reducing conditions showed that the high molecular weight SE-HPLC peak (peak 1) of the cooked products contained protein aggregates in addition to high molecular weight muscle proteins, while the second peak (peak 2) still contained aggregates and <50 kDa proteins. The protein aggregates composition was investigated by HPLC-tandem mass spectrometry. Different classes of proteins were identified and the cooked products showed a more complex composition and organization, according to the muscle structure and the technological procedures, respectively. The key role of actin in the building of the protein networks was also confirmed. The different multi-protein systems found in the cooked products suggest protein re-organization in heat-induced supramolecular structures, which might be responsible for the texture and the structural properties of the final products., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The influence of microwave treatment of hydrated durum wheat kernels of two different cultivars (cv Aureo and Sfinge), on wholemeal flour and pasta quality was addressed. Size exclusion-HPLC and electrophoresis analysis were used to investigate changes in the gluten network arrangement as affected by the microwave treatment. Rheological properties of dough, cooking quality and sensory properties of pasta were also assessed. Results suggested that the microwave treatment on hydrated durum wheat kernels blocks gluten protein conformation through SS bonds formation and the free -SH are no longer able to create a strong network during pasta processing, due to the conformational changes. Rheological study of dough confirmed that the modifications induced by microwave treatment greatly affected pasta making characteristics of wheat flour, with significant negative consequences on product quality, especially for pasta cooking quality. Pasta from treated durum wheat showed low sensory quality, mainly due to high bulkiness and adhesiveness., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Microwave based treatment (MWT) of wet wheat kernels induced a striking reduction of gluten, up to <20 ppm as determined by R5-antibodybased ELISA, so that wheat could be labeled as gluten-free. In contrast, analysis of gluten peptides by G12 antibody-based ELISA, mass spectrometry-based proteomics and in vitro assay with T cells of celiac subjects, indicated no difference of antigenicity before and after MWT. SDS-PAGE analysis and Raman spectroscopy demonstrated that MWT simply induced conformational modifications, reducing alcohol solubility of gliadins and altering the access of R5-antibody to the gluten epitopes. Thus, MWT neither destroys gluten nor modifies chemically the toxic epitopes, contradicting the preliminary claims that MWT of wheat kernels detoxifies gluten. This study provides evidence that R5-antibody ELISA alone is not effective to determine gluten in thermally treated wheat products. Gluten epitopes in processed wheat should be monitored using strategies based on combined immunoassays with T cells from celiacs, G12-antibody ELISA after proteolysis and proper molecular characterization., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

The experiment was carried out to evaluate the effects of a moderate level of flaxseed administration on milk coagulation properties and fatty acid profile of milk from two different breeds. The experiment was performed on 20 Italian Friesian cows and 20 Jersey cows divided into 2 groups of 10 animals each. The experimental diets were (1) a traditional diet (CON) administrated as unifeed and no supplemental fat and (2) a diet supplemented with 0·5 kg/d of whole flaxseed (FS). Cows were milked twice daily and milk yield was recorded. Milk samples were analysed at 1, 15, and 30 d of the experiment for composition, pH, and milk coagulation properties. To verify the effects of flaxseed administration on the coagulation properties of milk from Friesian and Jersey cows, an electrophoresis study on casein fractions was performed. Milk fatty acid profile can be improved by administrating a moderate level of flaxseed in the diet, however, milk fatty acid profile from Friesian and Jersey cows showed different contents of C18 : 1 trans-11, SFA and MUFA. The results demonstrated that milk coagulating ability can be increased by flaxseed administration in both breeds as a result of different aggregation of casein micelles.

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., (Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

To evaluate process-induced protein modifications in cooked ham and emulsion sausages, the proteomes of whole-cut (Parma and "Praga" cooked hams) and comminuted pork (mortadella and würstel) products were compared to raw pork using two-dimensional gel electrophoresis (2-DE) coupled to image analysis and mass spectrometry (MS). Other than heat-induced breakdown of part of the myosin heavy chains, the 2-DE pattern of cooked ham was substantially similar to that of raw pork. However, the MS-based analysis showed minor modifications, including the extensive oxidation of methionines. In contrast, likely due to emulsification, comminuted sausages were characterized by an abundant insoluble protein fraction (IPF). Interestingly, tropomyosin and myosin light chains in comminuted sausages were exclusively found in the IPF. Our results indicate that the protein aggregation systems of cooked hams and emulsion sausages reflect the processing conditions and are definitely different, the former being characterized mainly by disulphide bridges and the latter by additional covalent inter-protein links., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

The main challenge to probiotics, during their passage through the gastrointestinal tract, are the acidic gastric secretions of the stomach, and the bile salts released into the duodenum. The survival of the strains, in this phase, is strongly influenced by the food used for their delivery. This work is part of a project studying the development of novel food processes, based on the use of chestnuts from cultivar "Castagna di Montella". In detail, the effect of indigestible chestnut fiber and of chestnut extract on the viability of selected lactic acid bacteria strains was evaluated. Among 28 cultures, twelve strains were selected, on the basis of tolerance to low pH values and bile salts, and submitted to exposition to simulated gastric or bile juice in presence of chestnut extract with or without immobilization in chestnut fiber. The presence of chestnut extract proved to play a significant role on the gastric tolerance improvement of lactobacilli. The recorded protective effect could not be simply related to the starch or reducing sugars content. RP-HPLC demonstrated that in the chestnut flour, there are one or more hydrophobic peptides or oligopeptides, which specifically offer a marked resistance to simulated gastric juice, albeit present at low concentration. These beneficial effects proved to be dependent by the cultivar used to produce the flour., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Genetic manipulation of durum wheats by tobacco rab-1 genes influence the trafficking of gluten proteins through the secretory system by up- or down-regulating the transport step from the ER to the Golgi apparatus which may in turn modify functional performance of the grain. Gluten proteins were extracted from two genetically manipulated lines - Svevo B730 1-1 and Ofanto B688 1-2 - and their control lines and were analyzed by two dimensional gel electrophoresis. When the two-dimensional maps were compared by image analysis no significant differences between the GM line with an up-regulated trafficking containing the wild type tobacco rab1 (Svevo B730 1-1) and its control (Svevo control). By contrast, significant differences were found between the GM line with a down-regulated trafficking due to the tobacco rab1 mutant form (Ofanto B688 1-2) and its control (Ofanto control). Of the new protein spots detected in the down-regulated Ofanto B688 1-2 map, only a beta-amylase was identified. The remaining spots were susceptible to chymotripsin action but not to trypsin one, as in the case of the gluten protein. Rheological measurements showed that gluten quality was enhanced in the down-regulated Ofanto B688 1-2 without an increase in the amount of gluten. Proteomics is a useful and powerful tool for investigating protein changes in GMOs and in understanding events in food science and technology.