Gianfranco Mamone

Italian National Research Council | CNR · Institute of Food Sciences ISA

We investigated the digestive peptide profiles of pea and faba bean proteins to evaluate possible bioactive peptides (BAPs), and to test immune-modulating properties of selected peptides using in vitro cell model. More than 400 unique peptides were released upon gastrointestinal digestion of pea and faba bean proteins and identified by high-resolut...

Gluten proteins are the causative agents of celiac disease (CD), a lifelong and worldwide spread food intolerance, characterized by an autoimmune enteropathy. Gluten is a complex mixture of high homologous water-insoluble proteins, characterized by a high content of glutamine and proline amino acids that confers a marked resistance to degradation b...

Despite the physiological importance of the hydrolases from the intestinal brush border membrane (BBM), a step simulating the intestinal digestion has not been included yet in the harmonized protocols of in vitro digestion, due to commercial unavailability of these enzymes and lack of consensus for the conditions of use. The proper utilize of BBM r...

Every year, the coffee supply chain produces tons of waste materials from the processing of the coffee bean. Coffee silverskin (CS) is the only waste product from the coffee bean roasting phases but as roasting is a key step in obtaining the coffee beverage, there are tons of CS collected and thrown away by coffee roasting industries. This study is...

Gluten degrading enzymes, which are commonly referred to as “glutenases,” represent attractive candidates for the development of a pharmacological treatment of gluten related disorders, such as coeliac disease (CeD). Endoprotease-40 (E40), a novel glutenase secreted by the actinomycete Actinoallomurus A8 and recombinantly produced in S. lividans TK...

Alternative sources of edible proteins are required to feed the world’s growing population, such as Moringa oleifera leaves, a protein source with a balanced amino acid composition. Since Moringa leaf proteins is a novel food in the EU and UK, an assessment of their potential allergenicity of is required. Proteins from Moringa leaf powder were char...

Selected food proteins may represent suitable markers for assessing either the presence/absence of specific food ingredients or the type and intensity of food processes. A fundamental step in the quantification of any protein marker is choosing a proper protocol for solubilizing the protein of interest. This step is particularly critical in the cas...

Tritordeum results from the crossbreeding of a wild barley (Hordeum chilense) species with durum wheat (Triticum turgidum spp. turgidum). This hexaploid crop exhibits agronomic and rheological characteristics like soft wheat, resulting in an innovative raw material to produce baked goods. We applied a gel-based proteomic approach on refined flours...

Protein expression from the berry skin of four red grape biotypes with varying hybrid character was compared at a proteome-wide level to identify the metabolic pathways underlying divergent patterns of secondary metabolites. A bottom-up shotgun proteomics approach with label-free quantification and MaxQuant-assisted computational analysis was appli...

Omic sciences such as metabolomic, lipidomic, and proteomic were used for characterizing and identifying chemical compounds from seed germs such as flavonoids, lipids, and proteins of different species of Prosopis. Reversed phase-HPLC-diode array detector and nanoflow-HPLC coupled to tandem mass spectrometry (MS/MS) were used. Fatty acids of Prosop...

Pistachio nut (Pistacia vera) is an important tree nut cultivated widely worldwide and is commonly appreciated based on its organoleptic characteristics and nutritional healthfulness. Similar to other tree nuts, the consumption of pistachio nuts is associated with moderate or severe allergic reactions in sensitized/allergic individuals. In the pres...

The objective of this study was to characterize the insoluble and soluble dietary fiber present in Argentine and Italian artichoke (Cynara scolymus) by-products by comparing chemical analysis and ATR-FTIR spectroscopy. Non-edible bracts of Argentine (AR) and Italian (Benevento (BN) and Sicily (SC)) artichoke cultivars were employed. The soluble and...

Wheat consumption can represent one of the nutritional factors involved in the onset of diabetes. We specifically investigated the potential diabetogenic effects of Hammurabi, a T. monococcum wheat cultivar, in non-obese diabetic (NOD) mice and analysed the levels of resistant starch in pasta manufactured with Hammurabi after in vitro gastroduodena...

Scope: Several studies reported a role of amylase/trypsin-inhibitors (ATIs) of common wheat species in promoting immune reactions. Here, we investigated in celiac disease (CD), the immunogenic properties of ATIs from diploid compared to common hexaploid wheats after an in vitro proteolytic hydrolysis. Methods and Results ATIs purified from two lin...

The aim of this study was to evaluate the dynamics of proteolysis during dough fermentation started with different lactic acid bacteria species, through the identification of intermediate and small-sized peptides generated during fermentation. Single-strain cultures of Levilactobacillus brevis, Fructilactobacillus sanfranciscensis, Companilactobaci...

Infant Milk Formula (IMF) is designed as a breastmilk substitute to satisfy the nutritional requirements during the first months of life. This study investigates the effects of two IMF processing technologies on cow milk protein digestion using an infant static in vitro gastrointestinal model. The degree of protein hydrolysis at the end of the gast...

In this investigation, we reported the production of prototype breads from the processed flours of three specific Triticum turgidum wheat genotypes that were selected in our previous investigation for their potential low toxic/immunogenic activity for celiac disease (CD) patients. The flours were subjected to sourdough fermentation with a mixture o...

The lipid fraction of durum wheat (Triticum turgidum ssp. durum (Desf.) Husn.), cultivar Svevo, was characterized during grain maturation. Growing grain kernels were harvested at five key stages of development between 5 and 30 days post anthesis (dpa). Lipid evolution was assessed during kernel filling by gas chromatography-mass spectrometry (GC-MS...

Celiac disease (CeD) is an autoimmune enteropathy triggered by immunogenic gluten peptides released during the gastrointestinal digestion of wheat. Our aim was to identify T cell epitope-containing peptides after ex vivo digestion of ancestral (einkorn, spelt and emmer) and common (hexaploid) wheat (Fram, Bastian, Børsum and Mirakel) using human ga...

One hundred lactic acid bacteria from Italian sourdoughs were characterized for technological features and selected for their tolerance to stress conditions commonly encountered within the production of leavened fermented bakery products. Specifically, lactic acid bacteria belonging to the family Lactobacillaceae were evaluated for the capability t...

The protein/peptide composition of five beer kinds, including two experimental beer-like products brewed with einkorn (Triticum monococcum), a beer labeled as “gluten-free,” a traditional all-barley malt and a wheat (T. aestivum) containing beer, was characterized with HPLC-ESI MS/MS-based proteomics. To enlarge the characterization of the componen...

Heat treatments induce chemical/physical modifications, which may affect the stability to enzymatic digestion and consequently the allergenicity of food proteins to a varying extent, depending on the time/temperature regimen. Herein, we evaluated the stability to digestion of whole tree nut (walnuts, hazelnuts and almonds) allergens in a food diges...

Recent studies suggested that gliadin proteins from the ancient diploid einkorn wheat Triticum monococcum retained a reduced number of immunogenic peptides for celiac disease patients because of a high in vitro digestibility with respect to hexaploid common wheat. In this study, we compared the immunological properties of gliadins from two Triticum...

Scope Gluten from the diploid wheat Triticum monococcum (TM) has low content of immunostimulatory sequences and a high gastro‐intestinal digestibility. We analysed gluten‐reactive T cells elicited by diploid and hexaploid (Triticum aestivum‐TA) wheat in celiac disease (CD) patients upon a brief oral challenge. Methods and results Seventeen patient...

Prosopis alba and Prosopis nigra powders from pods were evaluated through chemical and mineral composition and other healthy food compounds such as polyphenols and fatty acids exhibiting antioxidant activity. P. nigra presented higher content of lipids and polyphenols leading to a higher antioxidant activity than P. alba. Although P. nigra presente...

Digestomics can be defined as the systematic characterization of the biomolecular complement produced by food digestion. Protein digestomics, that essentially corresponds to tracking the fate of dietary proteins in human body, assumes special relevance because food-derived peptides are virtually capable of inducing positive (health-promoting) or ne...

The wheat varietal selection undertaken by breeders in recent decades has been tailored mainly to improve technological and productivity-related traits; however, the latter has resulted in a considerable impoverishment of the genetic diversity of wheat-based products available on the market. This pitfall has encouraged researchers to revalue the na...

This study investigated the simultaneous impact of food matrix and processing on the food allergy eliciting capacity of peanuts in a physiologically relevant context. Whole raw and roasted peanuts were subjected to in vitro digestion combining the harmonized oral-gastric-duodenal digestion models with brush border membrane enzymes (BBM) to simulate...

Gluten proteins are the causative agent of Celiac Disease (CD), a life-long food intolerance characterized by an autoimmune enteropathy. Inadvertent gluten exposure is frequent even in celiac patients complying with a gluten-free diet, and the supplementation of exogenous gluten-digestive enzymes (glutenases) is indeed a promising approach to reduc...

Proteomics was exploited to assess the nature of possible traces of vomit found on the scene of an alleged sexual assault. In the case in point, a woman reported to the police to be raped five days before by a cousin of hers in his car. The woman declared she had vomited in the car before fainting definitely, due to alcohol or possible drugs covert...

This work aimed to study the opioid peptide β-casomorphin-7 (BCM7) degradation or stability during digestion using human gastrointestinal (GI) juices and porcine jejunal brush border membrane (BBM) peptidases. Synthetic BCM7 was subjected to in vitro digestion by GI fluids obtained from human volunteers for 180 min, and to downstream degradation wi...

Analyzing an in vitro gastroduodenal digest of whey proteins by HPLC coupled to high-resolution/high-sensitivity MS/MS, we sought to evaluate if state-of-art peptidomics provides comprehensive peptide coverage of a food “digestomes”. A multitude of small-sized peptides derived from both -lactalbumin and β-lactoglobulin as well as disulfide cross-l...

This work introduces a novel methodological approach to study both the geometry of complex protein networks and the nature of the interacting proteins. This approach is based on the high reactivity of Au⁺ ions on the surface of gold nanoparticles (AuNPs) towards thiols, that allows fast formation of a covalent bond between accessible protein thiols...

Nanoflow-HPLC-tandem mass spectrometry (MS/MS) was used to analyze the peptide fraction of breast milk samples collected from a single non-atopic donor on different days (10 samples) after receiving an oral load of cow's milk (by drinking 200 mL of bovine milk). In addition, breast milk was sampled from the same lactating mother over a 6-h period a...

Most of the bread on the market contains several additional ingredients compared to those used for traditional baking (flour, water, yeast, and salt). Modern bread is often baked using enzymatic dough improvers, which, as technological aids, can be omitted on the label. Baking mixes also can contain varying percentages of hard fat or their derivati...

Over the last years, great efforts have been devoted to develop effective gluten detoxification strategies with a consequent detrimental alteration of the technological properties as well. Obtaining low-gluten products without affecting the rheological properties of wheat could still be considered a new challenge to face. In this investigation, we...

By virtue of exclusive nutrient composition and nutritional properties, seed germ flours from both European carob (Ceratonia siliqua L.) and South American algarrobo (Prosopis alba and Prosopis nigra) or vinal (Prosopis ruscifolia) have potential as a high nutritional value and health-promoting ingredient for food formulations. In order to define t...

Flour from seed germ of European carob (Ceratonia siliqua) and South American algarrobo (Prosopis spp.) is a potential ingredient for health-promoting baked products. Herein, lipids from germ of three Argentinean Prosopis (P. alba, P. nigra, and P. ruscifolia) and one European carob species were characterized in detail, exploiting an array of up-to...

Hemp (Cannabis sativa L.), traditionally cultivated for industrial use and harvested for fibers and seeds, has raised much interest as a sustainable crop in the last years. Recently, hemp seeds and derived oil have started to be used in a variety of food products. Hemp-based food products are considered less allergenic than those from other edible...

In this work, we explored the “deep” seed peanut proteome by using both two dimensional electrophoresis (2-DE)-based analysis run under reducing and non-reducing condition (protein-centric) and LC-MS/MS gel-free proteomic (peptide-centric). The former approach allowed to identify high molecular weight disulfide-linked Ara h 1 and Ara h 3 heteroligo...

The present study aimed to obtain novel information regarding the pathways of a polyphenol-rich coffee silverskin extract (CSE) for inhibiting the formation of advanced glycation products (AGE). Chlorogenic acid (CGA) is the main phenol present in coffee beans. The contribution of CGA to the antiglycoxidative properties of CSE was also evaluated by...

A long-term psychiatric 40 years-old male patient was found dead at 9:00 a.m. in the clinic where he lived. Death was caused by traumatic injuries, which the sanitary staff imputed to a fall. Nurses declared that the patient refused having breakfast, whereas at autopsy the stomach contained 350 g of whitish semifluid material. Using both shotgun an...

Peanut allergy is one of the most widespread types of food allergies especially affecting developed countries. To reduce the risk of triggering allergic reactions, several technological strategies have been devised to modify or remove allergens from foods. Herein we investigated the combination of high temperature and pressure on the modulation of...

Triticum monococcum L. is one of the oldest ancestors of wheat. There is some evidence that einkorn encloses forms of gliadin-deriving peptides which may potentially exert a reduced toxicity to consumers with gluten-related disorders. Accordingly, ID331 and Monlis lines were comparatively investigated in this study. The biological effects of gastro...

Dietary fiber and whole grain foods may contribute to the regulation of appetite; however, evidence has produced inconclusive findings. The objective was to evaluate the effects of an experimental wholemeal pasta on appetite ratings, plasma concentrations of gastrointestinal hormones involved in appetite control, and postprandial glucose/insulin re...

In this study, we explored possibilities to develop low glycaemic-index foods from barley (Hordeum vulgare – Hv). Barley has a potential to suppress postprandial blood glucose levels, possibly because of its high content of β-glucan (BG). BG content is particularly high in Hordeum vulgare Subsp. spontaneum (Hs), which is the wild ancestor of cultiv...

Proteins determine several key sensory and technological properties of beer. Most of the protein components of beer are barley pathogenesis–related proteins, which endure the harsh brewing treatments. Due to their intrinsic resistance, they can also survive gastrointestinal digestion in humans, evoking immune responses in predisposed individuals. F...

Hazelnut (Corylus avellana) is widely used in food production in raw, roasted, salted, and paste form. Proteins are key components conferring favorable sensory, technological, and functional properties to hazelnut. For these reasons, the protein fraction of hazelnut has been the subject of extensive research over the last years. In addition, the co...

Enzymatic dough improvers (DIs) are increasingly used as baking co-adjuvants. Herein, an array of techniques, including Western blotting, PCR, electrophoresis-based and shotgun proteomics, was addressed to identify the enzymes in six commercial DI preparations. In particular, this work sought to exclude the possible undeclared use of amylolytic enz...

Stability to proteolytic degradation in the digestive tract is considered a general feature shared by most food allergens. Current digestibility models exclusively utilize purified allergen proteins, neglecting the relevant effects of matrix that occur for foodstuff systems. In the present study, we investigated digestion stability of the major pea...

Biological significance: A 2D-DIGE-based comparative analysis of the albumin/globulin fraction from durum wheat caryopses at six developmental stages was performed to describe the dynamic subproteomic changes associated with grain development. Quantitative variations of 217 differentially proteins demonstrated that highly affected are the function...

The time course of biosynthesis and accumulation of storage proteins in developing grain of durum wheat (Triticum turgidum ssp. durum (Desf.) Husn.) pasta-quality reference cv. Svevo was investigated at the protein level for the first time. Seeds were harvested at key kernel developmental stages, namely 3 (seed increase threefold in size), 5 (kerne...

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...

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...

Background: Extensively hydrolyzed casein formula (EHCF) has been proposed for the prevention and is commonly used for the treatment of cow's milk allergy (CMA). The addition of the probiotic Lactobacillus rhamnosus GG (LGG) to EHCF may induce faster acquisition of tolerance to cow's milk. The mechanisms underlying this effect are largely unexplor...

Heat shock 60 kDa protein 1 (HSP60) is a chaperone and stress response protein responsible for protein folding and delivery of endogenous peptides to antigen-presenting cells and also a target of autoimmunity implicated in the pathogenesis of atherosclerosis. By two-dimensional electrophoresis and mass spectrometry, we found that exposure of human...

Figure S1: Print-out of the query to the MASCOT interface (Matrix Science), concerning the mass spectrum (shown in Fig. 3) of the in-gel tryptic digestion products of spot number 3c from a preparative replica (200 μg protein load) of the gel shown in Fig. 2. Figure S2: Print-out of the query to the MASCOT interface (Matrix Science), concerning the...

The ancient diploid Triticum monococcum is of special interest as a candidate low-toxic wheat specie for celiac disease patients. Here, we investigated how an in vitro gastro-intestinal digestion, affected the immune toxic properties of gliadin from diploid compared to hexaploid wheat. Gliadins from Triticum monococcum, and Triticum aestivum cultiv...

Wheat beer is a traditional light coloured top-fermenting beer brewed with at least 50% of malted (e.g. German Weissbier) or unmalted (e.g. Belgian Witbier) wheat (Triticum aestivum) as adjunct to barley (Hordeum vulgare) malt. For the first time we explored the proteome of three Weissbier samples, using both two-dimensional electrophoresis (2DE)-b...

This study aimed to assess the impact of the "often neglected" intestinal brush border membranes (BBM) hydrolases on dietary peptides, exploring the possibility that the disintegration of proteins progressed in the small intestine up to a "core" of intrinsically stable oligopeptides, persisting independently on the up-stream breakdown. Samples of s...

The aim of the present study was to identify and characterize the celiacogenic/immunogenic proteins and peptides released during digestion of pasta (Triticum durum semolina). Cooked pasta was digested using a harmonized in vitro static model of the oral-gastro-intestinal digestion. The course of pasta protein digestion was monitored by SDS-PAGE, an...

In the present chapter, we describe the issue of food allergy and its distribution across the globe. Emphasis is also given to the celiac disease, a non-IgE-mediated food allergy. Then we pass to illustrate the prevalence of food allergy and the most common categories of allergenic foods currently regulated worldwide. In the second section, the foc...

Bresaola is traditionally produced by curing and air-drying entire anatomic cuts of lean bovine hindquarters. Because of its low-fat and high-protein content, Bresaola is increasingly appreciated for the nutritional properties and has become one of the best known and exported Italian meat products. Both sarcoplasmic and myofibrillar protein fractio...

A tryptic fragment (b5TR,NR), encompassing residues 2515-2750, was isolated from a low-iodine (0.26 % by mass) bovine thyroglobulin, by limited proteolysis with trypsin and preparative, continuous-elution SDS-PAGE. The fragment was digested with Asp-N endoproteinase and analyzed by reverse-phase HPLC electrospray ionization quadrupole time-of-fligh...

Biochemical aspects of the main components of wheat flour (proteins, carbohydrates, and lipids) are presented and discussed with a focus on those properties of relevance to the baking industry. As well as considering compositional and structural issues for each of the major macromolecules in flour, particular attention is paid to the interactions a...

Vinyl phenol reductase activity was assayed in extracts from 19 strains of Dekkera bruxellensis isolated from wine. In all strains, vinyl phenol reductase activity was insensitive to the presence/absence of 4-vinyl guaiacol, confirming that expression is not related to the presence of the substrate. D. bruxellensis CBS 4481 showed the highest vinyl...