Drivers of the In-Mouth Interaction between Lupin Protein Isolate and Selected Aroma Compounds: A Proton Transfer Reaction-Mass Spectrometry and Dynamic Time Intensity Analysis.

Plant proteins often carry off-notes, necessitating customized aroma addition. In vitro studies revealed protein-aroma binding, limiting release during consumption. This study employs in vivo nose space proton transfer reaction-time-of-flight-mass spectrometry and dynamic sensory evaluation (time intensity) to explore in-mouth interactions. In a lupin protein-based aqueous system, a sensory evaluation of a trained "green" attribute was conducted simultaneously with aroma release of hexanal, nonanal, and 2-nonanone during consumption. Results demonstrated that enlarging aldehyde chains and relocating the keto group reduced maximum perceived intensity (Imax_R) by 71.92 and 72.25%. Protein addition decreased Imax_R by 30.91, 36.84, and 72.41%, indicating protein-aroma interactions. Sensory findings revealed a perceived intensity that was lower upon protein addition. Aroma lingering correlated with aroma compounds' volatility and hydrophobicity, with nonanal exhibiting the longest persistence. In vitro mucin addition increased aroma binding four to 12-fold. Combining PTR-ToF-MS and time intensity elucidated crucial food behavior, i.e., protein-aroma interactions, that are pivotal for food design.

Unraveling the Role of Flavor Structure and Physicochemical Properties in the Binding Phenomenon with Commercial Food Protein Isolates.

Food protein-flavor binding influences flavor release and perception. The complexity of the binding phenomenon lies in the flavor and protein properties. Thus, molecular interactions between commercial whey- or plant-based protein isolates (PI) such as pea, soy, and lupin, with carbonyl and alcohol flavor compounds were assessed by static headspace (HS) GC-MS. HS results showed that not only the displacement of the carbonyl group from the inner part of the flavor structure toward the edge promoted binding up to 52.76% ± 4.65 but also the flavor's degree of unsaturation. Similarly, thermal treatment led to a slight increase in hexanal-protein binding because of possible protein conformational changes. Protein's residual fat (<1%) seemed insufficient to promote significant flavor binding to PI. Despite the complexity of commercial food protein isolates, the results displayed that binding is predominantly influenced by the flavor structure and physicochemical properties, with the protein source and residual fat playing a secondary role.

Lentil Protein and Tannic Acid Interaction Limits in Vitro Peptic Hydrolysis and Alters Peptidomic Profiles of the Proteins.

In this study, the nature of lentil protein-tannic acid (LPTA) interaction and its effect on in vitro pepsin digestion were investigated. LPTA mixtures containing 1% w/v LP and 0.001-0.5% TA were prepared and characterized in terms of particle size, thermal properties, and secondary and tertiary structures. A 20-fold increase in particle size was observed in LPTA0.5% compared to LP control (without TA), indicating aggregation. Static quenching of tryptophan residues within the protein hydrophobic folds was observed. Increasing TA levels also enhanced protein thermal stability. Over 50% reduction in free amino groups of LPTA 0.5%, relative to LP, was observed after pepsin digestion. Cleavage specificity of pepsin and peptidomic profile of LP were modified by the presence of TA in LPTA 0.5%. This study showed that 0.5% w/v TA induced protein aggregation and reduced LP digestibility by hindering the accessibility of pepsin to the protein network, thus modifying the profile of released peptides.

ß-Glucan Interaction with Lentil (Lens culinaris) and Yellow Pea (Pisum sativum) Proteins Suppresses Their In Vitro Digestibility.

In this study, ß-glucan interaction with lentil and yellow pea proteins and the effect on in vitro protein digestibility were investigated. Proteins were mixed with ß-glucan at mass ratios of 1:0.5, 1:1, and 1:2. The interaction between ß-glucan and the proteins was demonstrated by the decrease in transmittance and surface charge and the increase in particle size of the complexes. Bright-field microscopy showed the formation of aggregates between the biopolymers, although increased molecular size was not observed by discontinuous native polyacrylamide gel electrophoresis. Fluorescence microscopy indicated that ß-glucan formed aggregates with lentil proteins, while the interaction with yellow pea proteins appeared as distinct phases of protein within the ß-glucan network. The in vitro protein digestibility of lentil and pea protein decreased by 27.3 and 34.5%, respectively, in the presence of a ß-glucan mass ratio of 1:2. The findings confirm the possibility to modulate protein digestibility by changing the physical characteristics of a food matrix.

Inhibition of α-glucosidases by tea polyphenols in rat intestinal extract and Caco-2 cells grown on Transwell.

Inhibition of maltase, sucrase, isomaltase and glucoamylase activity by acarbose, epigallocatechin gallate, epicatechin gallate and four polyphenol-rich tea extract from white, green, oolong, black tea, were investigated by using rat intestinal enzymes and human Caco-2 cells. Regarding rat intestinal enzyme mixture, all four tea extracts were very effective in inhibiting maltase and glucoamylase activity, but only white tea extract inhibited sucrase and isomaltase activity and the inhibition was limited. Mixed-type inhibition on rat maltase activity was observed. Tea extracts in combination with acarbose, produced a synergistic inhibitory effect on rat maltase activity. Caco-2 cells experiments were conducted in Transwells. Green tea extract and epigallocatechin gallate show dose-dependent inhibition on human sucrase activity, but no inhibition on rat sucrase activity. The opposite was observed on maltase activity. The results highlighted the different response in the two investigated model systems and show that tea polyphenols are good inhibitors for α-glucosidase activity.

Tea polyphenols as a strategy to control starch digestion in bread: the effects of polyphenol type and gluten.

The inhibitory effect of tea polyphenols on starch digestibility can contribute to the control of the glycaemic index of starchy food. In this study, wheat bread and gluten-free bread were co-digested in vitro with different amounts of tea polyphenols. The kinetics of starch digestion and polyphenol bio-accessibility during in vitro digestion were monitored. The results showed that co-digestion of bread with tea polyphenols dose-dependently slowed the starch digestion kinetics and this effect is influenced by the types of polyphenols and the presence of gluten. The presence of gluten lowered the inhibitory efficacy of tannins on starch digestibility to 7.4% and 47.5% when 25 mg of tannins were co-digested with wheat bread and gluten-free bread, respectively. In contrast, the presence of gluten had little impact on the inhibitory efficacy of monomeric polyphenols. This study shows that the release of tea polyphenols in the digestive environment is a promising strategy for controlling the glycaemic index of starchy food and that monomeric and polymeric tea polyphenols differently affect starch digestion according to the presence of gluten.

Gastrointestinal Bioaccessibility and Colonic Fermentation of Fucoxanthin from the Extract of the Microalga Nitzschia laevis.

The extract of microalga Nitzschia laevis (NLE) is considered a source of dietary fucoxanthin, a carotenoid possessing a variety of health benefits. In the present study, the bioaccessibility and deacetylation of fucoxanthin were studied by simulated in vitro gastrointestinal digestion and colonic batch fermentation. In the gastric phase, higher fucoxanthin loss was observed at pH 3 compared to pH 4 and 5. Lipases are crucial for the deacetylation of fucoxanthin into fucoxanthinol. Fucoxanthinol production decreased significantly in the order: pure fucoxanthin (25.3%) > NLE (21.3%) > fucoxanthin-containing emulsion (11.74%). More than 32.7% of fucoxanthin and fucoxanthinol was bioaccessible after gastrointestinal digestion of NLE. During colon fermentation of NLE, a higher loss of fucoxanthin and changes of short-chain fatty acid production were observed but no fucoxanthinol was detected. Altogether, we provided novel insights on the fucoxanthin fate along the human digestion tract and showed the potential of NLE as a promising source of fucoxanthin.

Bioprocessing of common pulses changed seed microstructures, and improved dipeptidyl peptidase-IV and α-glucosidase inhibitory activities.

Type 2 diabetes mellitus (T2DM) is a leading cause of death globally. T2DM patients experience glucose intolerance, and inhibitors of dipeptidyl peptidase IV (DPP-IV) and α-glucosidase are used as drugs for T2DM management. DPP-IV and α-glucosidase inhibitors are also naturally contained in foods, but their potency can be affected by the food matrix and processing methods. In this study, germination and solid-state fermentation (SSF) were used to alter pulse seed microstructures, to convert compounds into more bioactive forms, and to improve their bioaccessibility. Germination substantially modified the seed microstructure, protein digestibility, contents and profiles of phenolic compounds in all the pulses. It also increased DPP-IV and α-glucosidase inhibitory activities in chickpeas, faba beans and yellow peas. Compared to germination, SSF with Lactobacillus plantarum changed the content and the profile of phenolic compounds mainly in yellow peas and green lentils because of greater disruption of the seed cell wall. In the same pulses, heat treatment and SSF of flour increased DPP-IV and α-glucosidase inhibitory activities. The results of this study suggest that germination and SSF with L. plantarum are effective and simple methods for modulating phenolic and protein profiles of common pulses and improve the action on DPP-IV and α-glucosidase.

The Effect of Sulforaphane on Glyoxalase I Expression and Activity in Peripheral Blood Mononuclear Cells.

Studies demonstrate that the potential health-beneficial effect of sulforaphane (SR), a compound formed in broccoli, is the result of a number of mechanisms including upregulation of phase two detoxification enzymes. Recent studies suggest that SR increases expression/activity of glyoxalase 1 (Glo1), an enzyme involved in the degradation of methylglyoxal, is major precursor of advanced glycation end products. Those compounds are associated with diabetes complications and other age-related diseases. In this study, the effect of SR on the expression/activity of Glo1 in peripheral blood mononuclear cells (PBMCs) from 8 healthy volunteers was investigated. PBMCs were isolated and incubated with SR (2.5 µM-concentration achievable by consuming a broccoli portion) for 24 h and 48 h. Glo1 activity/expression, reduced glutathione (GSH), and glutathione-S-transferase gene expression were measured. Glo1 activity was not affected while after 48 h a slight but significant increase of its gene expression (1.03-fold) was observed. GSTP1 expression slightly increased after 24 h incubation (1.08-fold) while the expressions of isoform GSTT2 and GSTM2 were below the limit of detection. GSH sharply decreased, suggesting the formation of GSH-SR adducts that may have an impact SR availability. Those results suggest that a regular exposure to SR by broccoli consumption or SR supplements may enhance Glo1.

Isothiocyanates from Brassica Vegetables-Effects of Processing, Cooking, Mastication, and Digestion.

The formation of health-beneficial isothiocyanates (ITCs) from glucosinolates depends on a wide variety of plant-intrinsic factors (e.g., concentration of glucosinolates, activity of myrosinase, and specifier proteins) and on a multitude of extrinsic postharvest factors such as the conditions used during industrial processing, domestic preparation, mastication, and digestion. All of these factors contribute to a large variability in the formation of ITCs (and other breakdown products), as well as their intake and absorption upon consumption of Brassica vegetables. This uncertainty in ITC intake and absorption is a barrier for the determination of an optimal Brassica vegetable consumption pattern. In this review, the intrinsic and extrinsic factors that affect the formation, intake, and absorption of ITCs are described according to the most recent findings. The focus of this review includes the hydrolysis reaction mechanisms, the elucidation of the primary factors that play a role in the hydrolysis reaction, the influence of processing and cooking conditions, the effect of chewing, and the roles of the gastric and upper intestinal phases, including the effect of the meal composition (e.g., the effect of other meal compounds present during digestion) on the potential formation of ITCs.

Role of the food matrix and digestion on calculation of the actual energy content of food.

The energy content of food is calculated on the basis of general factors for fat, protein, and carbohydrates. These general factors were derived by W.O. Atwater in the late 19th century, while additional factors for dietary fiber, polyols, and organic acids were introduced more recently. These factors are applied indiscriminately to all types of foods, yet the same nutrient may be digested to different extents to generate energy, depending on the characteristics of the food matrix, the processing methods applied to foods, and the meal composition. As a consequence, the actual energy content of food may differ from what is theoretically calculated with the Atwater factors. In this review, the relationship of macronutrient digestibility with food structure, macronutrient structure, and food composition is examined, and the implications for the amount of energy achievable through diet are highlighted. Estimates of the discrepancy between calculated energy content and actual energy content are provided for different diets. The findings may have implications for consumer purchasing decisions as well as for the design of dietary interventions.

Bioavailability of Isothiocyanates From Broccoli Sprouts in Protein, Lipid, and Fiber Gels.

SCOPE: Optimization of bioavailability of dietary bioactive health-beneficial compounds is as important as increasing their concentration in foods. The aim of this study is to explore the change in bioavailability of isothiocyanates (ITCs) in broccoli sprouts incorporated in protein, fiber, and lipid gels. METHODS AND RESULTS: Five participants took part in a cross-over study and collected timed urine samples up to 24 h after consumption of proteins, dietary fibers, and lipid gels containing broccoli sprouts powder. Sulforaphane and iberin metabolites were determined in the urine samples. Samples in which sulforaphane and iberin were preformed by myrosinase led to a higher bioavailability of those compounds. Compared to the control broccoli sprout, incorporation of sprouts in gels led to lower bioavailability for preformed sulforaphane and iberin (although for sulforaphane the lower bioavailability was not significantly different) whereas for the gels rich in their precursors, glucoraphanin and glucoiberin, the opposite trend was observed (although not significantly different). CONCLUSION: This explorative study suggests that ITCs bioavailability can be modulated by food structure and composition and further and deeper investigations are needed to develop food products that lead to an optimized ITCs bioavailability.

Brassica rapa hairy root extracts promote skin depigmentation by modulating melanin production and distribution.

BACKGROUND: Skin whitening products, used for ages by Asian people for cultural and esthetic purposes, are very popular nowadays in Western countries as well, where the need to inhibit skin spots after sun exposure has become not only a cosmetic but also a health-related issue. Thus, the development of effective and safe depigmenting agents derived from natural products gets continuous attention by cosmetic brands and consumers. OBJECTIVES: The aim of this study was to determine the effects of two preparations, obtained from the hairy root cultures of the species Brassica rapa, on melanogenesis and the expression of the extracellular matrix proteins involved in a correct pigment distribution. METHODS: The two preparations, obtained by water-ethanol extraction and by digestion of cell-wall glycoproteins of the root cells, were chemically characterized and tested on skin cell cultures and on human skin explants to investigate on their dermatological activities. RESULTS: Both the extracts were able to decrease melanin synthesis pathway in melanocytes and modulate the expression of genes involved in melanin distribution. One of the extracts was also effective in inducing the expression of laminin-5 and collagen IV, involved into the maintenance of tissue integrity. The two extracts, when tested together on human skin explants, demonstrated a good synergic hypopigmenting activity. CONCLUSIONS: Taken together, the results indicate that the extracts from B. rapa root cultures can be employed as cosmetic active ingredients in skin whitening products and as potential therapeutic agents for treating pigmentation disorders.

Modeling food matrix effects on chemical reactivity: Challenges and perspectives.

The same chemical reaction may be different in terms of its position of the equilibrium (i.e., thermodynamics) and its kinetics when studied in different foods. The diversity in the chemical composition of food and in its structural organization at macro-, meso-, and microscopic levels, that is, the food matrix, is responsible for this difference. In this viewpoint paper, the multiple, and interconnected ways the food matrix can affect chemical reactivity are summarized. Moreover, mechanistic and empirical approaches to explain and predict the effect of food matrix on chemical reactivity are described. Mechanistic models aim to quantify the effect of food matrix based on a detailed understanding of the chemical and physical phenomena occurring in food. Their applicability is limited at the moment to very simple food systems. Empirical modeling based on machine learning combined with data-mining techniques may represent an alternative, useful option to predict the effect of the food matrix on chemical reactivity and to identify chemical and physical properties to be further tested. In such a way the mechanistic understanding of the effect of the food matrix on chemical reactions can be improved.

Stir-Frying of Chinese Cabbage and Pakchoi Retains Health-Promoting Glucosinolates.

Stir-frying is a cooking method, originating from Asia, in which food is fried in small amount of very hot oil. Nowadays in many other parts of the world stir-frying is a very popular method to prepare vegetables, because it is fast and fried vegetables are tasty. However, the retention of phytochemicals like the health-beneficial glucosinolates in Brassica vegetables is less explored for stir-frying in comparison to other cooking methods. This study investigates the retention of glucosinolates in Chinese cabbage (Brassica rapa ssp. pekinensis) and pakchoi (Brassica rapa ssp. chinensis) as affected by stir-frying at various cooking durations and temperatures. Stir-frying experiments were performed at set pan temperatures ranging from 160 to 250 °C for a duration of 1 to 8 min. Results showed that aliphatic glucobrassicanapin is the most abundant glucosinolate identified in fresh Chinese cabbage and pakchoi, contributing for 48 and 63% of the total glucosinolate content, respectively, followed by glucoiberin and gluconapin. Stir-frying retains the glucosinolates even at the highest temperature applied. Such retention is explained by the quick inactivation of the glucosinolate-hydrolytic enzyme myrosinase during the first minutes of frying, and by the thermal stability of the glucosinolates at those temperature/time conditions. Moreover, due to the absence of a separate water phase, leaching losses did not occur, in contrast to what is observed when boiling Brassica vegetables. These results show that stir-frying may be a suitable health-beneficial cooking option that prevents the loss of glucosinolates.

Food as Pharma? The Case of Glucosinolates.

BACKGROUND: Glucosinolates (GLSs) are dietary plant secondary metabolites occurring in the order Brassicales with potential health effects, in particular as anti-carcinogenic compounds. GLSs are converted into a variety of breakdown products (BPs) upon plant tissue damage and by the gut microbiota. GLS biological activity is related to BPs rather than to GLSs themselves. METHODS: we have reviewed the most recent scientific literature on the metabolic fate and the biological effect of GLSs with particular emphasis on the epidemiological evidence for health effect and evidence from clinical trials. An overview of potential molecular mechanisms underlying GLS biological effect is provided. The potential toxic or anti-nutritional effect has also been discussed. RESULTS: Epidemiological and human in vivo evidence point towards a potential anti-cancer effect for sulforaphane, indole-3-carbinol and 3,3-diindolylmethane. A number of new human clinical trials are on-going and will likely shed further light on GLS protective effect towards cancer as well as other diseases. BPs biological effect is the results of a plurality of molecular mechanisms acting simultaneously which include modulation of xenobiotic metabolism, modulation of inflammation, regulation of apoptosis, cell cycle arrest, angiogenesis and metastasis and regulation of epigenetic events. BPs have been extensively investigated for their protective effect towards cancer but in recent years the interest also includes other diseases. CONCLUSION: It appears that certain BPs may protect against and may even represent a therapeutic strategy against several forms of cancer. Whether this latter effect can be achieved through diet or supplements should be investigated more thoroughly.

Optimizing isothiocyanate formation during enzymatic glucosinolate breakdown by adjusting pH value, temperature and dilution in Brassica vegetables and Arabidopsis thaliana.

Consumption of glucosinolate-rich Brassicales vegetables is associated with a decreased risk of cancer with enzymatic hydrolysis of glucosinolates playing a key role. However, formation of health-promoting isothiocyanates is inhibited by the epithiospecifier protein in favour of nitriles and epithionitriles. Domestic processing conditions, such as changes in pH value, temperature or dilution, might also affect isothiocyanate formation. Therefore, the influences of these three factors were evaluated in accessions of Brassica rapa, Brassica oleracea, and Arabidopsis thaliana. Mathematical modelling was performed to determine optimal isothiocyanate formation conditions and to obtain knowledge on the kinetics of the reactions. At 22 °C and endogenous plant pH, nearly all investigated plants formed nitriles and epithionitriles instead of health-promoting isothiocyanates. Response surface models, however, clearly demonstrated that upon change in pH to domestic acidic (pH 4) or basic pH values (pH 8), isothiocyanate formation considerably increases. While temperature also affects this process, the pH value has the greatest impact. Further, a kinetic model showed that isothiocyanate formation strongly increases due to dilution. Finally, the results show that isothiocyanate intake can be strongly increased by optimizing the conditions of preparation of Brassicales vegetables.

The kinetic of key phytochemical compounds of non-heading and heading leafy Brassica oleracea landraces as affected by traditional cooking methods.

BACKGROUND: Kales are often a key ingredient of traditional foods, containing high amounts of indolic glucosinolates (precursors of indole-3-carbinol and ascorbigen), carotenoids and phenolics. The present trend to associate traditional foods crops with health-promoting properties suggested to investigate the degradation kinetic of three Brassica oleracea landraces' phytochemicals subjected to boiling, steaming and stir-frying. RESULTS: Boiling led to substantial losses due to leaching. Glucosinolates followed a second-order degradation kinetic (20% of their initial values after 10 min in Nero di Toscana). Phenolic content in leaves + cooking water remained unchanged, whereas their antioxidant capacity was reduced. Carotenoid content increased during the first minutes of boiling. Steaming showed the highest retention of phytochemicals, with often zero-order degradation kinetic, having however a strong effect on colour. Stir-frying produced high losses for all measured compounds; also, ß-carotene reduced its content to 10-23% independently of variety. Conversion values for indole-derived compounds ranged from non-detectable to 23.5%. CONCLUSION: Variety strongly affected observed degradation rates because of a different glucosinolate composition and leaf structure. With this research, more information has been gained on the degradation kinetic of B. oleracea landraces' phytochemical compounds upon cooking, highlighting the possibility of improving bioactive component retention. © 2016 Society of Chemical Industry.