1,720,978 research outputs found
Molecular tools for the protection of agrobiodiversity and promotion of sustainable agriculture
Full text linkThe protection and conservation of grain agrobiodiversity represent milestones in modern and sustainable agriculture and food waste reduction. Molecular tools play a vital role in understanding grain traits, although their integration into agricultural practices remains limited. Our study, part of the CROPDIVA project, focuses on developing biochemical methods to assess grain quality and select high-performing varieties for different applications. Our research aimed at establishing a high-throughput screening pipeline for CROPDIVA varieties, enhancing agricultural diversity and promoting new supply chains. Diversified agricultural products contribute to soil enrichment and dietary improvement.
We conducted comprehensive bromatological and phytochemical characterizations of CROPDIVA samples, evaluating protein, lipid, humidity and fiber content, enzymatic starch assays, and bioactive and nutritional features such as total polyphenols, anti-nutritional factors, and total antioxidant capacity. Furtherly, selected CROPDIVA grain varieties were evaluated by adopting an in vitro digestion models to assess dry matter and protein digestibility, and nutrient bioavailability of selected nutrient compound.
In conclusion, integrating minor crops into feed enhances agricultural sustainability, benefits human nutrition, and supports ecosystem resilience. Our findings underscore the importance of agrobiodiversity in fostering resilient agricultural systems that meet global food security challenges
Nuovi approcci metodologici allo studio delle interazioni tra proteine in matrici complesse: applicazioni al network proteico di cereali
No full textLe proprietà del glutine sono influenzate dalla natura e dal numero di interazioni che avvengono tra le proteine coinvolte. Le caratteristiche intrinseche di gliadine e glutenine rendono tuttavia difficile una chiara e completa identificazione degli aspetti molecolari che sono alla base della formazione del reticolo glutinico. La descrizione delle interazioni proteina-proteina e delle caratteristiche geometriche del reticolo risultante potrebbe consentire di ottenere una migliore comprensione dell’evoluzione del processo di reticolazione, del ruolo dei trattamenti tecnologici, e del rapporto struttura funzione per questa matrice complessa.
Lo scopo del presente lavoro è di sviluppare approcci basati sulla chimica delle proteine per ottenere informazioni in merito ai determinanti molecolari che sono alla base delle interazioni tra le proteine di una matrice alimentare, con particolare riferimento al ruolo dei gruppi tiolici cisteinici nella formazione di legami covalenti e nei processi di “disulfide exchange”.
Un primo approccio prevede di valutare la reattività dei gruppi tiolici delle diverse frazioni proteiche del frumento tramite l’utilizzo di marcatori fluorescenti. La separazione elettroforetica mono- e bi-dimensionale delle proteine marcate permette di individuare nel pattern proteico totale le proteine aventi gruppi tiolici reattivi (non impegnati in ponti disulfuro e accessibili al solvente). L’approccio descritto è stato utilizzato (i) per evidenziare differenze strutturali nel pattern proteico di linee isogeniche con un modificato rapporto amilosio/amilopectina e (ii) per valutare modificazioni nella mappa, indicativa delle natura delle interazioni intra- e inter-molecolari, in seguito a impastamento e cottura.
Un secondo approccio sfrutta la capacità di nanoparticelle d’oro di formare legami covalenti con i gruppi tiolici cisteinici. L’utilizzo di nanoparticelle di diverse dimensioni (comprese tra i 2 e i 200 nm) consente di valutare l’accessibilità dei gruppi tiolici in funzione delle caratteristiche geometriche del reticolo glutinico di materie prime e semilavorati ottenuti da semole e farine. Effettuando la marcatura anche in presenza di agenti dissocianti sarà possibile ottenere informazioni sul ruolo delle interazioni idrofobiche nella stabilizzazione del reticolo glutinico
From proteins to bread: novel tools for the molecular description of protein-protein interactions (and their evolution) in baked products
No full textCereal-based matrices are deeply studied and characterized from a rheological and technological point of view. Although protein-protein interactions at the base of gluten formation are well known (i.e. disulfide exchange reactions and hydrophobic interactions), the peculiar chemistry of the proteins involved in the formation of the network - gliadins and glutenins - is still a source of discussion. The present work aims at developing new molecular approaches suitable to describe the “thiolome” of the system, in order to provide information on gluten protein structure and protein-protein interactions during processing. The thiolomic approach relies on labeling accessible free cysteine thiols with a specific iodoacetamide derivative fluorescent probe, in the presence of different destructuring agents (SDS and urea). Labeled proteins are separated in mono- and/or bi-dimensional electrophoresis. Image analysis is used to compare the fluorescent emission map and the total protein pattern to detect changes in the distribution of free thiols among the various proteins. The comparison of the information obtained from the thiolomic approach with those obtained from other approaches, such as front-face fluorescence and differential solubility, can provide various insights into the evolution of the protein structure of the matrix. Preliminary results on a soft wheat-based baked product (bread), show that changes in the thiols pattern during kneading - i.e., when disulfide exchange and rearrangement of hydrophobic patches occur - are comparatively modest with respect to those observed after cooking, a step characterized by temperature-induced structural rearrangements and by water transfer from proteins to starch
A strategy to implement the circular economy paradigm using okara extracts fractions and biochemical approach in agri-tech innovation
Full text linkThe significant environmental and economic impacts of waste and byproducts from food chains are well-documented. However, these byproducts still contain numerous valuable molecules with potential applications and bioactivities in various fields. Moreover, the environmental impact and economic aspects related to the usage of agri-food by-products and food waste represent a cornerstone in advancing the circular economy [1]. Okara, a byproduct from soy, Glycine max, generated from soymilk production, is gaining attention due to its abundance and the numerous possible applications in the agri-food sector. This work focused on the fractionation of okara proteins through green, sustainable, water-based extraction processes [2] and the enzymatic production of peptides. We tested different peptide fractions for their potential inhibitory capacity against pathogenic fungi (i.e. Fusarium graminearum) and bio-stimulatory activities tested on plant systems. Fractionating waste is crucial for a comprehensive analysis and a deep understanding of the activities of the single peptide obtained through enzymatic digestion. This approach facilitates comprehensive investigations into residual proteins' molecular and biochemical properties and valuable compounds, which can be studied using both in vitro and in vivo methodologies
How sustainable is sustainability? Recovery of bioactives from food processing waste
Full text linkThe modern agri-food industry, crucial for global nutrition, generates significant by-products often considered waste, posing economic and environmental challenges. By-products are secondary products unintentionally obtained from the manufacturing process, while waste refers to inefficient activities that add no value. Waste is generated throughout the entire food lifecycle, from production to distribution. They are frequently incinerated or sent to landfills, causing air, water, and soil pollution. Agri-food waste can alternatively be used as animal feed, yet both waste and by-products contain valuable molecules for scientific research and sustainable solutions in a circular economy. Agri-food by-products contain compounds for biostimulation, green pesticides, and nutrition, requiring chemical-physical treatments to realize their potential. Therefore, revalorization of agri-food waste biomass creates high-value products and addresses waste accumulation, underscoring the importance of economical evaluation in this process.
Despite studies on pure isolated bioactive compounds, significant gaps persist in understanding their interactions with matrices, influenced by their supramolecular characteristics, which affect their behavior during isolation. Matrix effects impede the exploration of biological activities such as biodefense or biostimulation, stemming from molecular changes that occur during processing. Overcoming these challenges requires innovative biochemical strategies and enzymatic treatments.
The recovery of bioactives from food processing waste highlights critical challenges for the agri-food sector. Private and public companies in nutraceuticals, food packaging, and agriculture stand to benefit significantly from these innovations. Crucially, EU policies play a pivotal role in managing food waste and by-products, advancing a sustainable circular economy by reducing waste and optimizing resource efficiency
Combining the reactivity of gold nanoparticles and proteomics approaches to address protein-protein interactions in food proteins
No full textThe thiol-binding capacity of Au nanoparticles (AuNPs, 20 nm) was used to test accessibility of protein thiols in wheat flour, where proteins interact through both hydrophobic interactions and disulfide bonds. At difference with soluble thiol reagents, the AuNPs size limits their penetration in protein networks. Also, the stability of the thiol ligation to AuNPs simplifies removal of unbound proteins and recovery of AuNPs (and of covalently bound proteins) through centrifugation, making it possible to identify AuNPs-bound proteins through MS/MS and limited proteolysis. Low concentrations of detergents allowed controlled breakdown of hydrophobic interactions and allowed to distinguish binding of AuNPs to insoluble network-forming proteins rather than to soluble ones. Both glutenins (having free cysteines and intramolecular disulfides) and gliadins (no free thiols and only intramolecular disulfides) were found to be covalently bound to AuNPs, proving that the two protein classes form intermolecular disulfides already in the flour. These results pave the way to further studies on the time course and the molecular determinants of protein-protein interactions during deposition in the seed or on the role of some proteins or protein classes in gluten formation. Use of AuNPs of different size should also make it possible to assess the geometry and the structure of the protein network in foods, either wheat-based or not
Matrix effect : a challenge to enhance the bioavailability of functional bioactive compounds derived from okara, the byproduct of soymilk production
Full text linkThe rising interest towards sustainable food production has led to the exploitation of byproducts. Despite the fact they are considered wastes, byproducts are still containing a high quantity of nutrients and bioactive compounds. In the last few years, the effects of these molecules on health and food processing have been deeply explored. However, the “matrix effect” exerted by the supramolecular organization hasn’t aroused the same interest. Highly processed byproducts present this structural reorganization that could affect the release of compounds during their digestion, absorption and function on the target.
This work aims to release the protein fraction and other interesting molecules from okara using several extraction methods to overcome the matrix effect. Okara, the soymilk processing byproduct, has a protein content representing about 30% of its dry weight and it is largely produced generating an environmental and economic problem. Different approaches have been performed to increase the bioavailability and bioactivity of functional components. The fractions obtained in this way have been tested on human colorectal adenocarcinoma cells (CaCo2). In particular, cytotoxicity activity has been analyzed by testing different concentrations. Also, the protective effects on cell inflammation have been studied on cells, and the amount of IL1β expressed has been measured. Since the expression levels of NFκB mirror the expression of chemokine IL8, the expression of IL8 was quantified by qRT-PCR to evaluate the anti-inflammatory effect of peptides. The study has shown that each treatment has a different impact on the release of compounds, which also influences their bioactivity.
Acknowlegments: This study was carried out within the Agritech National Research Center and received funding from the European Union NextGenerationEU (PIANO NAZIONALE DI RIPRESA E RESILIENZA (PNRR) – MISSIONE 4 COMPONENTE 2, INVESTIMENTO 1.4 – D.D. 1032 17/06/2022, CN00000022)
Molecular information for addressing climate changes (and consumers’ demands): wheat proteins
No full textClimate change and shifting patterns in the consumer’s demand represent major challenges for the agrifood chain. In the case of grain-based products, consumers would appreciate staple foods (pasta and bread) with traditional sensory features but without their nutritional drawbacks, such as the high glycemic index. Protein networks play a crucial role in defining the sensory characteristics in cereal-based products by forming both covalent and non-covalent interactions. Process-induced structural changes (and their kinetics) affect the organization of proteins in the network, resulting in macroscopic changes. However, other components in the matrix may affect a number of steps in the process, including competition for water at various steps of processing. A detailed knowledge of these effects is also pertinent to the production of food enriched with nutritionally relevant ingredients - including fiber, phenolics, and other bioactives - obtained from agrifood by-products.
This work addresses the pattern of protein-protein interactions in isogenic bread wheat lines, that included a control line (WT), a waxy (i.e. no amylose) line (WX), and a high-amylose line (HA), along with a commercial bread wheat flour. Protein profiles were assessed by mono- and bi-dimensional electrophoresis. The chemical nature of protein-protein interactions was assessed by studying the sensitivity of the water-insoluble proteins to the sequential addition of salts, chaotropes, and disulfide reductants. Properties of hydrophobic patches on the protein surface were assessed by measuring binding of ANS (1,8 aniline naphthalene sulfonate) by front-face fluorescence. A “thiolomics” approach based on labeling of thiols with 5-(iodoacetamide) fluorescein (IAF) was used to assess the accessibility of protein thiols in the original materials and in protein fractions solubilized in suitable solvent systems.
The protein patterns in the isogenic wheats were identical, but different from that in the commercial flour in both gluten components (monomeric gliadins and polymeric glutenins). In spite of the identical protein profile, the sensibility of the insoluble protein aggregates to both chaotropes and disulfide reductants in the isogenic samples increased in the order: WT<
Assessing the geometrical features of protein-protein interactions in gluten
No full textThis study exploits the reactivity of the surface of Au nanoparticles (AuNPs) towards thiols to test accessibility of protein thiols in protein in durum wheat semolina, where proteins interact through both hydrophobic interactions and disulfide bonds. After removal of loosely bound proteins, proteins covalently bound the AuNPs were identified by MS/MS after limited proteolysis.
We found it necessary to loosen hydrophobic interactions in the network to grant accessibility to AuNPs (20 nm average size). This required the use of low molarity urea or detergents, and the use of appropriate controls to verify binding of AuNPs to proteins that may be solubilized under the same mild-denaturing conditions.
Both glutenins (containing free cysteines and intramolecular disulfides) and gliadins (having no free thiols and only intramolecular disulfides) were found to be covalently bound to AuNPs, proving that the two protein classes are interacting in the grain through intermolecular disulfides. Current tests are aimed at verifying how these results can be affected by previous blockage of some of the accessible free thiols on these proteins prior to exposure to AuNPs, and are carried out by using specific fluorescent probes. This combination of approaches should provide further insights into the nature of the involved proteins and the geometry of their interactions
The biochemistry for the valorization of agri food waste and by-products: the case of Okara for sustainable AgriTech Applications in Circular Economy
Full text linkFood chains play a significant environmental and economic role for their consequences regarding the large amounts of waste and byproducts produced. Recently, the monetization of matrices obtained from agrifoods wastes has been a crucial step forward for the circular economy concept. Biochemically, such byproducts are very interesting because of their content in proteins, peptides, and other highvalue molecules like polyphenols. Therefore, these matrices will have high biotechnological potential for different fields and applications. Okara, the byproduct of soy (Glycine max) [1], has garnered agro and biotechnological interest because of its versatility,
sustainability, and abundance [2]. In the present work, we developed different approaches for the recovery and purification of Okara’s peptides, which will be valorized
for their agrotechnological potential (e.g. biodefense activity). Sustainable extraction methodologies and hydrolytic enzymes were used to overcome the matrix effects that hinder extraction yields. Biochemical methodologies including sequencing and spectroscopy were used for the molecular characterizations of the peptides purified in the different fractions. The selected fractions were tested with different in vivo systems to assess their biological activities
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