Effect of milk serum proteins on aggregation, bacteriostatic activity and digestion of lactoferrin after heat treatment
Xiong, Ling ; Boeren, Sjef ; Vervoort, Jacques ; Hettinga, Kasper - \ 2020
Food Chemistry 337 (2020). - ISSN 0308-8146
Bacteriostatic activity - In vitro digestion - Lactoferrin - Milk serum proteins - Thermal aggregation
To establish the effect of the presence of milk serum proteins on heat-induced changes to lactoferrin, lactoferrin alone, and lactoferrin mixed with either milk serum or β-lactoglobulin was heated at 65 °C, 70 °C and 75 °C for 30 min. After heating, the effect of milk serum proteins on aggregation of lactoferrin was characterized, after which the effect of such aggregation on digestion and bacteriostatic capacity of lactoferrin were determined. The presence of milk serum proteins accelerated the aggregation of lactoferrin during heating through thiol/disulphide interchange. Lactoferrin also formed disulphide-linked aggregates when it was heated with β-lactoglobulin. Protein aggregates formed at 75 °C were much more resistant to infant digestion, causing decreased peptide release from lactoferrin. Heating lactoferrin and milk serum proteins together accelerated the loss of bacteriostatic activity upon heating. In conclusion, heat-induced aggregation of lactoferrin with milk serum proteins affected both its digestion and its bacteriostatic activity.
Heat treatment of β-lactoglobulin affects its digestion and translocation in the upper digestive tract
Deng, Ying ; Govers, Coen ; Tomassen, Monic ; Hettinga, Kasper ; Wichers, Harry J. - \ 2020
Food Chemistry 330 (2020). - ISSN 0308-8146
Caco-2 monolayer - Heat treatment - Hydrophobicity - Immune system - In vitro digestion - Translocation - β-Lactoglobulin
Heat treatment is a commonly applied unit operation in the processing of β-lactoglobulin containing products. This does, however, influence its structure and thereby impacts its activity and digestibility. We describe how various heat-treatments of β-lactoglobulin change the digestibility using a modified version of the current consensus INFOGEST protocol. Additionally, protein was investigated for its translocation over the intestinal epithelial barrier, which would bring them in contact with immune cells. The extent of gastric digestibility was higher when the protein structure was more modified, while the influence of glycation with lactose was limited. Translocation studies of protein across Caco-2 cell monolayers showed a lower translocation rate of protein heated in solution compared to the others. Our study indicates that structural modifications after different heat-treatments of β-lactoglobulin increase in particular gastric digestibility and the translocation efficiency across intestinal epithelial cells.
Interaction of bread and berry polyphenols affects starch digestibility and polyphenols bio-accessibility
Kan, Lijiao ; Oliviero, Teresa ; Verkerk, Ruud ; Fogliano, Vincenzo ; Capuano, E. - \ 2020
Journal of Functional Foods 68 (2020). - ISSN 1756-4646
Berry polyphenols - Bread - In vitro digestion - Polyphenol bio-accessibility - Starch digestibility
In this study, the effect of berry polyphenols on starch digestion was tested in vitro both by co-digestion of berry extract with bread or by fortifying bread with berry extract. Results show that the co-digestion of bread with berry extracts significantly reduce the rate and extent of starch digestion. Sixty one percent of starch digestion is inhibited by co-digesting 1 g of raspberry extract with 4 g of the bread. The inhibition obtained by co-digesting berry extracts and bread is much higher than the inhibition obtained by digesting berry-fortified bread. Interactions of polyphenols with matrix reduce polyphenols bio-accessibility, thus reducing the amount of polyphenols available for α-amylase inhibition. The interaction of polyphenols and starch seems also a crucial mechanism for the inhibition of starch digestion. This study shows that the co-ingestion of berry polyphenols with bread is a promising strategy to reduce glycaemic index of starchy food.
Heat-induced changes in microstructure of spray-dried plant protein isolates and its implications on in vitro gastric digestion
Rivera del Rio, Andrea ; Opazo-Navarrete, Mauricio ; Cepero-Betancourt, Yamira ; Tabilo-Munizaga, Gipsy ; Boom, Remko M. ; Janssen, Anja E.M. - \ 2020
Food Science and Technology = Lebensmittel-Wissenschaft und Technologie 118 (2020). - ISSN 0023-6438
Gastric digestion - In vitro digestion - Pea protein isolate - Protein digestibility - Soy protein isolate - Spray-dried protein isolates
The quickly expanding field of plant-based food, generally uses protein concentrates or isolates as protein source. It is however not clear to what extent the intensive processing of these raw materials affects their digestibility. We here report on the in vitro gastric digestibility of the structures present in unheated and heated dispersions of spray-dried protein isolates of soybean and yellow pea. Unheated dispersions consist primarily of insoluble individual spray-dried particles, agglomerates of these and only a small fraction of soluble protein. Pepsin activity was followed in real-time through microscopic observations, showing the disassociation of agglomerates and inward-breakdown of individual particles, which are otherwise stable at gastric pH and ionic strength. This demonstrates that solubility is not necessarily an incentive for gastric protein digestion. Heating does not significantly affect the overall digestibility of protein isolate dispersions. Nevertheless, heating disrupts the structure of spray-dried particles, increasing the amount of smaller and better digestible particles that remain suspended after centrifugation. Conversely, heat-induced aggregates remain in the pellet and are up to 50% less digestible than their unheated counterparts. This impaired digestibility is counterbalanced by a reduced proportion of poorly-digestible species in the full system (up to 11% for soy and 23% for pea).
Effect of domestic cooking methods on protein digestibility and mineral bioaccessibility of wild harvested adult edible insects
Manditsera, Faith A. ; Luning, Pieternel A. ; Fogliano, Vincenzo ; Lakemond, Catriona M.M. - \ 2019
Food Research International 121 (2019). - ISSN 0963-9969 - p. 404 - 411.
Boiling - Crickets - Eulepida mashona - Henicus whellani - In vitro digestion - Iron bioaccessibility - Mineral availability - Roasting
Wild harvested edible insects are characterised by high protein and mineral contents with potential to contribute substantially to nutrition security. However, nutritional content is only beneficial when proteins are digestible and minerals bioaccessible. This study determined the effects of domestic processing on protein digestibility and mineral bioaccessibility of two wild harvested insect species: Eulepida mashona (beetle) and Henicus whellani (cricket). Samples of both insects were subjected to boiling, roasting, or combined boiling and roasting, imitating the way insects are traditionally prepared in Zimbabwe. Moreover, they were in vitro digested according to INFOGEST protocol. Boiling of both insects resulted in loss of protein as it leached into the boiling water. The raw insects had a higher protein in vitro digestibility than the boiled and roasted insects, and the maximal decrease in protein digestibility was around 25% for twice boiling of the beetles and for boiled and roasted crickets. For both insect species, boiling resulted in non-significant loss of iron and zinc. Iron was the least bioaccessible mineral in both insects, based on the concentrations of soluble mineral measured by ICP-AES. However, beetles had a much higher iron bioaccessibility (30.7%) as compared to crickets (8.11%). Interestingly, boiling resulted in about 50% decrease in iron and zinc bioaccessibility in both species while roasting did not. The reduced protein digestibility and mineral accessibility with processing can be explained by protein modification and interactions of minerals with other food components, such as chitin and phytochemicals. Because of the reduction in protein digestibility and mineral accessibility during boiling, roasting should be favoured over boiling and in any case short boiling time is recommended.
Blood–brain barrier transport and neuroprotective potential of blackberry-digested polyphenols : an in vitro study
Figueira, Inês ; Tavares, Lucélia ; Jardim, Carolina ; Costa, Inês ; Terrasso, Ana P. ; Almeida, Andreia F. ; Govers, Coen ; Mes, Jurriaan J. ; Gardner, Rui ; Becker, Jörg D. ; McDougall, Gordon J. ; Stewart, Derek ; Filipe, Augusto ; Kim, Kwang S. ; Brites, Dora ; Brito, Catarina ; Brito, M.A. ; Santos, Cláudia N. - \ 2017
European Journal of Nutrition (2017). - ISSN 1436-6207 - p. 1 - 18.
Blackberry - Brain endothelial cells - In vitro digestion - Microarrays - Neuronal cells
Purpose: Epidemiological and intervention studies have attempted to link the health effects of a diet rich in fruits and vegetables with the consumption of polyphenols and their impact in neurodegenerative diseases. Studies have shown that polyphenols can cross the intestinal barrier and reach concentrations in the bloodstream able to exert effects in vivo. However, the effective uptake of polyphenols into the brain is still regarded with some reservations. Here we describe a combination of approaches to examine the putative transport of blackberry-digested polyphenols (BDP) across the blood–brain barrier (BBB) and ultimate evaluation of their neuroprotective effects. Methods: BDP was obtained by in vitro digestion of blackberry extract and BDP major aglycones (hBDP) were obtained by enzymatic hydrolysis. Chemical characterization and BBB transport of extracts were evaluated by LC–MSn. BBB transport and cytoprotection of both extracts was assessed in HBMEC monolayers. Neuroprotective potential of BDP was assessed in NT2-derived 3D co-cultures of neurons and astrocytes and in primary mouse cerebellar granule cells. BDP-modulated genes were evaluated by microarray analysis. Results: Components from BDP and hBDP were shown to be transported across the BBB. Physiologically relevant concentrations of both extracts were cytoprotective at endothelial level and BDP was neuroprotective in primary neurons and in an advanced 3D cell model. The major canonical pathways involved in the neuroprotective effect of BDP were unveiled, including mTOR signaling and the unfolded protein response pathway. Genes such as ASNS and ATF5 emerged as novel BDP-modulated targets. Conclusions: BBB transport of BDP and hBDP components reinforces the health benefits of a diet rich in polyphenols in neurodegenerative disorders. Our results suggest some novel pathways and genes that may be involved in the neuroprotective mechanism of the BDP polyphenol components.
Food-grade Micro-encapsulation Systems that May Induce Satiety via Delayed Lipolysis: A Review
Corstens, M.N. ; Berton-Carabin, C.C. ; Vries, R.J. de; Troost, F.J. ; Masclee, A.A.M. ; Schroen, C.G.P.H. - \ 2017
Critical Reviews in Food Science and Nutrition 57 (2017)10. - ISSN 1040-8398 - p. 2218 - 2244.
In vitro digestion - Ileal brake - emulsion - Food - obesity
The increasing prevalence of overweight and obesity requires new, effective prevention and treatment strategies. One approach to reduce energy intake is by developing novel foods with increased satiating properties, which may be accomplished by slowing down lipolysis to deliver substrates to the ileum, thereby enhancing natural gut-brain signalling pathways of satiety that are normally induced by meal intake. To develop slow release food additives, their processing in the gastrointestinal tract has to be understood; therefore, we start from a general description of the digestive system and relate that to in vitro modelling, satiety and lipolytic mechanisms. The effects of physicochemical lipid composition, encapsulation matrix and interfacial structure on lipolysis are emphasized. We give an overview of techniques and materials used, and discuss partitioning, which may be a key factor for encapsulation performance. Targeted release capsules that delay lipolysis form a real challenge because of the high efficiency of the digestive system; hardly any proof was found that intact orally ingested lipids can be released in the ileum and thereby induce satiety. We expect that this challenge could be tackled with structured o/w-emulsion-based systems that have some protection against lipase, e.g., by hindering bile salt adsorption and/or delaying lipase diffusion.
Protein identification and in vitro digestion of fractions from Tenebrio molitor
Yi, Liya ; Boekel, M.A.J.S. van; Boeren, Sjef ; Lakemond, Catriona M.M. - \ 2016
European Food Research and Technology 242 (2016). - ISSN 1438-2377 - p. 1285 - 1297.
In vitro digestion - Insect protein - LC–MS/MS - Protein identification - Tenebrio molitor
The nutritional value of insect protein is evaluated not only in amino acid composition, but also in protein digestibility. The general amino acid composition of Tenebrio molitor has been reported before, but limited knowledge is available on its digestibility. The objective of this study was to investigate in vitro protein digestibility of whole T. molitor larvae, a water-soluble fraction (supernatant) and water-insoluble fractions (pellet and residue), and to identify which proteins were present in the fractions studied. The digestibility of the supernatant fraction (~80 %) was much higher than that of pellet (~50 %) and residue (~24 %) after in vitro gastroduodenal digestion as was determined using the o-phthaldialdehyde (OPA) method. More proteins were digested after pepsin/pancreatin digestion than after only pepsin digestion. The most abundant proteins in the supernatant were hemolymph protein (~12 kDa), alpha-amylase (~50 kDa, a putative allergen), and muscle proteins (e.g. actin 30–50 kDa) in the pellet fraction as determined from LC–MS/MS and SDS-PAGE. In conclusion, the proteins in the soluble fraction that contained hemolymph proteins were more easily digestible than the insoluble, muscle protein-containing fractions.
Influence of processing and in vitro digestion on the allergic cross-reactivity of three mealworm species
Broekhoven, Sarah Van; Bastiaan-Net, Shanna ; Jong, N.W. De; Wichers, H.J. - \ 2016
Food Chemistry 196 (2016). - ISSN 0308-8146 - p. 1075 - 1083.
Crustaceans - Food allergy - Food processing - House dust mite - Ige cross-reactivity - In vitro digestion - Mealworms
Edible insects are currently being evaluated as an alternative and more sustainable protein source for humans. The introduction of new food sources can lead to development of novel allergies. Because in the Western world, insects are unlikely to be consumed raw, it is important to know how processing and in vitro digestion might influence their allergenicity. Three edible mealworm species (Tenebrio molitor, Zophobas atratus and Alphitobius diaperinus) subjected to processing and in vitro digestion were analysed for IgE cross-reactivity. Immunoblot and MALDI-MS/MS analyses revealed that IgE from crustaceans or House dust mite (HDM) allergic patients showed cross-reactivity to mealworm tropomyosin or α-amylase, hexamerin 1B precursor and muscle myosin, respectively. Heat processing as well as in vitro digestion did diminish, but not eliminate, HDM or tropomyosin IgE cross-reactivity. Results show that individuals allergic to HDM or crustaceans might be at risk when consuming mealworms, even after heat processing.
The harmonized INFOGEST in vitro digestion method : From knowledge to action
Egger, Lotti ; Ménard, Olivia ; Delgado-Andrade, Cristina ; Alvito, Paula ; Assunção, Ricardo ; Balance, Simon ; Barberá, Reyes ; Brodkorb, Andre ; Cattenoz, Thomas ; Clemente, Alfonso ; Comi, Irene ; Dupont, Didier ; Garcia-Llatas, Guadalupe ; Lagarda, María Jesús ; Feunteun, Steven Le; Janssen Duijghuijsen, Lonneke ; Karakaya, Sibel ; Lesmes, Uri ; Mackie, Alan R. ; Martins, Carla ; Meynier, Anne ; Miralles, Beatriz ; Murray, B.S. ; Pihlanto, Anne ; Picariello, Gianluca ; Santos, C.N. ; Simsek, Sebnem ; Recio, Isidra ; Rigby, Neil ; Rioux, Laurie Eve ; Stoffers, Helena ; Tavares, Ana ; Tavares, Lucelia ; Turgeon, Sylvie ; Ulleberg, E.K. ; Vegarud, G.E. ; Vergères, Guy ; Portmann, Reto - \ 2016
Food Research International 88 (2016)Par B. - ISSN 0963-9969 - p. 217 - 225.
Dairy proteins - Harmonized IVD protocol - In vitro digestion - Inter-laboratory trial - Mass spectrometry - Peptides
Within the active field of in vitro digestion in food research, the COST Action INFOGEST aimed to harmonize in vitro protocols simulating human digestion on the basis of physiologically inferred conditions. A harmonized static in vitro digestion (IVD) method was recently published as a primary output from this network. To validate this protocol, inter-laboratory trials were conducted within the INFOGEST network. A first study was performed using skim milk powder (SMP) as a model food and served to compare the different in-house digestion protocols used among the INFOGEST members. In a second inter-laboratory study applying the harmonized protocol, the degree of consistency in protein hydrolysis was investigated. Analysis of the hydrolyzed proteins, after the gastric and intestinal phases, showed that caseins were mainly hydrolyzed during the gastric phase, whereas β-lactoglobulin was, as previously shown, resistant to pepsin. Moreover, generation of free amino acids occurred mainly during the intestinal phase.The study also showed that a few critical steps were responsible for the remaining inter-laboratory variability. The largest deviations arose from the determination of pepsin activity. Therefore, this step was further clarified, harmonized, and implemented in a third inter-laboratory study.The present work gives an overview of all three inter-laboratory studies, showing that the IVD INFOGEST method has led to an increased consistency that enables a better comparability of in vitro digestion studies in the future.