Impact of caseins and whey proteins ratio and lipid content on in vitro digestion and ex vivo absorption

Caseins and whey proteins are known as ‘slow’ and ‘fast’ proteins, respectively, based on their amino acid absorption rate. However, there is limited understanding of the mechanisms controlling their behaviour during gastro-intestinal transit. A protein model system (8% total protein) with varying casein:whey protein ratios (0:100, 20:80, 50:50 and 80:20) were subjected to in vitro gastro-intestinal digestion using a semi-dynamic gastric model, a static intestinal model and an ex vivo absorption model (Ussing chambers). The casein-rich (≥ 50%) samples showed the formation of solid coagula that were persistent throughout gastric digestion, which caused a delay in nutrient emptying, slower digestion and leucine absorption kinetics. In contrast, whey proteins formed more soluble aggregates during the gastric phase, which led to faster gastric emptying, rapid intestinal hydrolysis, and higher and faster leucine absorption. This work shows the key role of the gastric restructuring for the overall digestive mechanism and kinetics of food, in particular proteins

Dairy food structures influence the rates of nutrient digestion through different in vitro gastric behaviour

peer-reviewedThe purpose of this study was to investigate in vitro the extent to which specific food structures alter gastric behaviour and could therefore impact on nutrient delivery and digestion in the small intestine. Results obtained from a specifically developed gastric digestion model, were compared to results from a previous human study on the same foods. The semi-dynamic model could simulate the main gastric dynamics including gradual acidification, lipolysis, proteolysis and emptying. Two dairy-based foods with the same caloric content but different structure were studied. The semi-solid meal comprised a mixture of cheese and yogurt and the liquid meal was an oil in water emulsion stabilised by milk proteins. Our findings showed similar gastric behaviour to that seen previously in vivo. Gastric behaviour was affected by the initial structure with creaming and sedimentation observed in the case of liquid and semi-solid samples, respectively. Lipid and protein digestion profiles showed clear differences in the amount of nutrients reaching the simulated small intestine and, consequently, the likely bioaccessibility after digestion. The semi-solid sample generated higher nutrient released into the small intestine at an early stage of digestion whereas nutrient accessibility from liquid sample was delayed due to the formation of a cream layer in the gastric phase. This shows the strong effect of the matrix on gastric behaviour, proteolysis and lipolysis, which explains the differences in physiological responses seen previously with these systems in terms of fullness and satiety.This work has funded by the Irish Dairy Levy Research Trust (project number MDDT6261). Ana-Isabel Mulet-Cabero was funded under Teagasc Walsh Fellowship scheme and BBSRC in the UK (grant BB/J004545/1)

Standardization of in vitro digestibility and DIAAS method based on the static INFOGEST protocol

Resumen de la conferencia invitada presentada en el 7th International Conference on Food Digestion. Cork, Irlanda. 3-5 Mayo (2022

Structural mechanism and kinetics of in vitro gastric digestion are affected by process-induced changes in bovine milk

Bovine milk is commonly exposed to processing, which can alter the structure, biochemical composition, physico-chemical properties and sensory quality. While many of these changes have been studied extensively, little is known about their effect on digestive behaviour. In this study, heat treatments of pasteurisation at 72 °C for 15 s or Ultra-High-Temperature (UHT) treatment at 140 °C for 3 s and homogenisation at pilot-plant scale were applied to whole milk. The gastric behaviour was investigated using a recently developed semi-dynamic adult in vitro model. The emptied digesta were analysed to assess the nutrient delivery kinetics, changes in microstructure and protein digestion. All samples showed protein aggregation and coagulum formation within the first 15 min of gastric digestion at which time the pH ranged from 5.5 to 6. Homogenised samples creamed regardless of heat treatment, whereas all non-homogenised samples exhibited sedimentation. The consistency of the coagulum of the heated samples was more fragmented compared to those of the non-heated samples. Rheological analysis showed that the higher the temperature of the heat treatment, the softer the obtained coagulum and the higher the protein hydrolysis at the end of digestion. The study also confirmed that gastric emptying of caseins from milk is delayed due to coagulation in the stomach, while β-lactoglobulin was emptied throughout the gastric phase, except for UHT-treated milk. The gastric behaviour also had an impact on the lipid and protein content of the emptied chyme. The homogenised samples seemed to release more nutrients at the end of gastric digestion

Dairy food structures influence the rates of nutrient digestion through different in vitro gastric behaviour

The purpose of this study was to investigate in vitro the extent to which specific food structures alter gastric behaviour and could therefore impact on nutrient delivery and digestion in the small intestine. Results obtained from a specifically developed gastric digestion model, were compared to results from a previous human study on the same foods. The semi-dynamic model could simulate the main gastric dynamics including gradual acidification, lipolysis, proteolysis and emptying. Two dairy-based foods with the same caloric content but different structure were studied. The semi-solid meal comprised a mixture of cheese and yogurt and the liquid meal was an oil in water emulsion stabilised by milk proteins. Our findings showed similar gastric behaviour to that seen previously in vivo. Gastric behaviour was affected by the initial structure with creaming and sedimentation observed in the case of liquid and semi-solid samples, respectively. Lipid and protein digestion profiles showed clear differences in the amount of nutrients reaching the simulated small intestine and, consequently, the likely bioaccessibility after digestion. The semi-solid sample generated higher nutrient released into the small intestine at an early stage of digestion whereas nutrient accessibility from liquid sample was delayed due to the formation of a cream layer in the gastric phase. This shows the strong effect of the matrix on gastric behaviour, proteolysis and lipolysis, which explains the differences in physiological responses seen previously with these systems in terms of fullness and satiety

Effects of grain source and processing methods on the nutritional profile and digestibility of grain amaranth

Amaranth grain is reputed to have a high nutritional value, and as a plant, be tolerant to adverse weather conditions. This suggests that grain amaranth could be useful in tackling malnutrition and the growing burden of cardiometabolic diseases. However, there is insufficient knowledge at present about how the nutrient composition and digestibility of amaranth grain varies with growing environment, crop genotype, and post-harvest processing. We investigated the effect of the source and processing of amaranth grains on the digestibility of protein and lipid present in the grains. There was variation in the composition and digestibility of raw grains from different sources, indicating a role of genotype and/or growing environment which warrants further investigation. The greatest differences in digestibility were measured between the different processing techniques. This indicates that efforts to increase the cultivation and consumption of grain amaranth need to be supported by education about effective processing and preparation techniques

Standardization of in vitro digestibility and DIAAS method based on the static INFOGEST protocol

Background: The FAO recommends the digestible indispensable amino acid score (DIAAS) as the measure for protein quality, for which the true ileal digestibility needs to be assessed in humans or pigs. However, due to high costs and ethical concerns, the FAO strongly encourages as well the development of validated in vitro methods, which complement the in vivo experiments. Method: Recently, an in vitro workflow, based on the validated static INFOGEST protocol, was developed and compared towards in vivo data. In parallel to the validation with in vivo data, the repeatability and reproducibility of the in vitro protocol were tested in an international ring trial (RT) with the aim to establish an international ISO standard method within the International Dairy Federation (IDF). Five different dairy products (skim milk powder, whole milk powder, whey protein isolate, yoghurt, and cheese) were analyzed in 32 different laboratories from 18 different countries, across 4 continents. Results: in vitro protein digestibilities based on Nitrogen, free R-NH2, and total amino acids as well as DIAAS values were calculated and compared to in vivo data, where available. Conclusion: The in vitro method is suited for quantification of digestibility and will be further implemented to other food matricesinfo:eu-repo/semantics/publishedVersio

Impact of pectin on the gastric digestibility of sodium caseinate

Trabajo presentado a las III Jornadas Científicas CIAL Fórum, celebradas del 22 al 23 de noviembre de 2018 en el Instituto de Investigación en Ciencias de la Alimentación (CIAL).Pectin is an anionic polysaccharide used in food applications which, due to its physical and chemical properties can modify the absorption of nutrients. The fact that there is a protection by polysaccharides to protein digestion can be very important when considering the absorption of nutrients and allergens in the small intestine such is the case of casein. One possible mechanism is through electrostatic binding to proteins, depending on the pH. Thus, the adsorption of pectin to casein can take place at pH 3.5-5.0 by the electrostatic attraction between the carboxylate group of pectin and cationic residue of casein. Hardly any investigation has been done on the interaction casein/pectin during digestion of the protein. In this work, we studied the effect of citrus pectin on the gastric digestion of sodium caseinate in a semi-dynamic gastric model. Different mixtures of caseinate and pectin (1/0.08-1/3.4) were assayed and the changes in the protein fraction due to the gastric digestion were followed by SDS-PAGE. After digestion, diffuse bands corresponding to the formation of peptides, whose intensity was greater when the ratio caseinate/pectin was lower, were observed. In addition, a slight displacement to areas of lower molecular mass was also detected. This was probably due to a protective effect of pectin, mainly after 1 h of gastric digestion. The best ratio caseinate/pectin was 1/0.08, since the other values (1/0.85 and 1/3.4) gave rise to lower initial pH values (4.83 and 4.38, respectively) near to the optimum pH of pepsin (1.5-4.5) that could have favored the digestion. With the aim to investigate if other parameters such as the molecular mass could affect, citrus pectin treated with pectinolytic enzymes such as Viscozyme® and Glucanex®200 G were used. SDS-PAGE analyses indicated that, besides pH that affect the electrostatic bindings between the polysaccharide and protein, the degree of polymerization of pectin plays also an important role in the casein digestion by steric hindrance. Although more investigation should be carried out to go more insight in the involved mechanism, the obtained results underline the protection of pectin on the digestion of sodium caseinate, particularly evident at times lower than two hours.This work has been funded by MINECO of Spain: Project AGL2014-53445-R. MV thanks Salvador de Madariaga Program for the economic support of this project.Peer reviewe

In vitro digestion of prebiotic carbohydrates added to milk using a rat intestinal enzyme preparation

Resumen del trabajo presentado a la 10th NIZO Dairy Conference: "Innovations in Dairy Ingredients", celebrada en Papendal (Netherlands) del 1 al 3 de octubre de 2017.Given the huge interest towards the gastrointestinal function and prebiotics, new oligosaccharides with improved properties are being explored, such as those derived from lactulose (OsLu), recently characterized and evaluated. Assays in rats have showed their significantly higher resistance when compared with galactooligosaccharides (GOS), probably ascribed to the terminal monosaccharide (fructose vs glucose). However, no in vitro studies have been done on the digestion of OsLu within a real food. Thus, our aim was to study the effect of a rat small intestinal extract (RSIE) on the hydrolysis of lactose, lactulose (Duphalac®), commercial GOS and OsLu in samples of milk after their gastric digestion in a semi-dynamic model. OsLu and commercial prebiotics were added to milk at the prebiotic dose (5%) and the products were firstly subjected to a gastric digestion. The resulting mixtures were freeze-dried. Samples (5 mg) were later added to 100 mg of RSIE together with 1 mL of distilled water, the reaction was kept at 37' for 2 h taking aliquots after 0, 0.5, 1 and 2 h. Trimethyl-silylated oximes of carbohydrates were determined by GC-FID. Results showed that, under the assayed conditions, lactulose was more resistant than lactose due to the presence of fructose in the molecule. Hydrolysis of oligosaccharides (di-, tri- and tetrasaccharides) from OsLu was lower (15%) as compared to present oligosaccharides in GOS (35%) after 2 h, probably due to the linkage type and the terminal monosaccharide. These finding support a prebiotic role of galactosyl-fructose derivatives (OsLu) in foods, remarking their resistance to digestion not only as a free-ingredient but also as a part of a real food.Peer Reviewe

β-glucan release from fungal and plant cell walls after simulated gastrointestinal digestion

β-glucans are soluble fibre found in plant and fungal cell walls, which are linked with reduced serum lipid levels. This study investigates the β-glucan release from two fungal samples (mycoprotein and white button mushroom) compared with two plant samples, oat and barley bran. Our results indicate no β-glucan release after the simulated gastrointestinal digestion of the uncooked fungal samples. In contrast, following cooking and digestion, β-glucans were released from the mycoprotein matrix. Pancreatic enzymes facilitated the solubilisation of β-glucans. Protein and carbohydrate digestion appeared independent of β-glucan release in both uncooked and cooked samples. Conversely, the viscosity increased in the samples that showed a significant release of β-glucans at the end of gastrointestinal digestion. Structural analysis by scanning electron microscopy showed changes in the uncooked and cooked samples before and after digestion. This study shows for the first time the different behaviour in β-glucan release from fungal and plant samples