Improved estimation of in vitro protein digestibility of different foods using size exclusion chromatography (original) (raw)
Related papers
Protein digestion of different protein sources using the INFOGEST static digestion model
2020
In vitro digestion systems are valuable tools for understanding and monitoring the complex behavior of food degradation during digestion, thus proving to be good candidates for replacing in vivo assays. The aim of the present work was to study protein hydrolysis in a selection of different protein sources using the harmonized INFOGEST static protocol: three isolated proteins (collagen, zein, and whey protein) and five foods (sorghum flour, wheat bran cereals, peanuts, black beans, and pigeon peas). The proteins of all the substrates were analyzed by SDS-PAGE and HPLC-MS/MS. Individual amino acid composition was analyzed by high-performance liquid chromatography (HPLC). EAA/NEAA (essential amino acids/ nonessential amino acids) ratios in the substrates from low to high were as follows: wheat bran cereals, peanuts, collagen, zein, whey protein, sorghum, pigeon peas, and black beans. The results revealed sorghum, whey protein, and zein as good sources of BCAA. In all substrates, no intact protein from the substrates was visually detected by SDS-PAGE after the intestinal phase of in vitro digestion with the INFOGEST protocol. However, digestion-resistant peptides were detected in all substrates after the intestinal digestion phase. Protein hydrolysis was high in whey protein isolate and pigeon pea and low for wheat bran cereals and bovine collagen.
Digestibility of proteins from different sources
The Annals of the University Dunarea de Jos of Galati Fascicle VI – Food Technology, 2020
In vitro protein digestibility can be useful for estimating protein nutritional quality. Eight protein sources were chosen to study in vitro protein digestibility, namely: sodium caseinate, whey, mushrooms, pea, soy, oat, hemp and sea buckthorn. Trypsin was used to achieve the enzymatic hydrolysis and the pH-drop technique was employed to determine in vitro protein digestibility. Substrate hydrolysis was rapidly initiated after enzyme addition in the case of pea protein, sodium caseinate, soy and whey proteins, whereas a slower pH decrease was registered in the case of mushroom proteins. The in vitro protein digestibility decreased in the following order: pea (87.2%) > soy (85.9%) > whey (85.6%) > caseinate (85%) > hemp (78.5%) > oat (77.5%) > sea buckthorn (76.2%) > mushrooms (68.2%). The sea buckthorn and mushrooms samples with the lowest protein contents (15.6% and 18.3%, respectively) had the lowest protein digestibility. However, no correlation between protein content and protein digestibility was found.
Journal of AOAC INTERNATIONAL, 2005
The kinetics of peptide release during in vitro digestion of 4 protein sources (casein, cod protein, soy protein, and gluten) were investigated. Samples were sequentially hydrolyzed with pepsin (30 min) and pancreatin (2, 4, or 6 h) in a dialysis cell with continuous removal of digestion products. Nondialyzed digests were fractionated by ion-exchange chromatography and ultrafiltration. Animal proteins were digested at a greater rate than plant proteins. Target amino acids of specific enzymes appeared more rapidly in the dialyzed fractions when compared to other amino acids. Throughout the hydrolysis, nondialyzed digests contained a higher proportion of peptide mixtures with basic-neutral properties. Except for gluten, peptide fractions with molecular weights that exceeded 10 kDa (basic-neutral, BN > 10) were rapidly hydrolyzed during the first 2 h of pancreatin digestion. The kinetics of release and the composition of peptide fractions were different when the protein sources were...
Journal of Food Biochemistry, 1992
7he effects of various food components on the in vitro digestibility of proteins, measured by the immobilized digestive enzyme assay (IDEA) system, were investigated. The digestibility of unprocessed and processed sodium caseinate and soybean protein was examined. Varying concentrations of sucrose (0, 5, 10 and 20%) or starch (0, 0.5, I and 2 %) had no sign$cant effects on digestibility of protein. Similarly, the presence of emulsij?ed (I X lecithin) vegetable oil (0, 0.5, 1 and 2%) did not aflect digestibility. fierefore, these common ingredients of foods did not affect the extent of hydrolysis of the protein samples under the conditions of the IDEA method, suggesting that this assay is suitable for use with complex foods.
Refining in vitro digestibility assays: Fractionation of digestible and indigestible peptides1
Journal of Animal Science, 2004
Typically, in vitro methods used for estimating the amount of ileal digestible AA do not exhaustively digest samples, and arbitrary methods for separating digestible from indigestible protein are used. This may lead to over-or underestimation of digestibility coefficients. A method that exhaustively digests proteins using pepsin and pancreatin was developed, and the first objective of this research was to confirm that exhaustive digestion was indeed appropriate and to determine the fractionation method for separating digestible from indigestible proteins. For this, three homoarginine-labeled animal proteins were prepared. Samples were subsequently digested in vivo and in vitro to determine which fraction should be considered indigestible, and in vitro followed by in vivo to determine whether the extent of digestion in vivo was improved by predigestion. In vivo, soluble but unabsorbed peptides were smaller than 1 kDa, suggesting that the size of soluble peptides is not what prevents their absorption. Thus, all in vitro-soluble proteins should be considered digestible. In vitro, 88 ± 3% of the soluble peptides were smaller than 1 kDa, with the remainder between 1 and 5 kDa, suggesting that in
Refining in vitro digestibility assays: fractionation of digestible and indigestible peptides
Journal of animal science, 2004
Typically, in vitro methods used for estimating the amount of ileal digestible AA do not exhaustively digest samples, and arbitrary methods for separating digestible from indigestible protein are used. This may lead to over- or underestimation of digestibility coefficients. A method that exhaustively digests proteins using pepsin and pancreatin was developed, and the first objective of this research was to confirm that exhaustive digestion was indeed appropriate and to determine the fractionation method for separating digestible from indigestible proteins. For this, three homoarginine-labeled animal proteins were prepared. Samples were subsequently digested in vivo and in vitro to determine which fraction should be considered indigestible, and in vitro followed by in vivo to determine whether the extent of digestion in vivo was improved by predigestion. In vivo, soluble but unabsorbed peptides were smaller than 1 kDa, suggesting that the size of soluble peptides is not what prevents...
ELECTROPHORESIS, 2014
The present work describes a foodomics protocol coupling an in vitro static simulation of digestion to a combination of omics techniques, to grant an overview of the protein digestibility of a meat‐based food, namely Bresaola. The proteolytic activity mediated by the digestive enzymes is evaluated through Bradford and SDS‐PAGE assays, combined to NMR relaxometry and spectroscopy, to obtain information ranging from the microscopic to the molecular level, respectively. The simple proteomics tool adopted here points out that a clear increase of bioaccessible proteins occurs in the gastric phase, rapidly disappearing during the following duodenal digestion. However, SDS‐PAGE and the Bradford assay cannot follow the fate of the digested proteins when the products are sized <5 kDa. Conversely, NMR spectroscopy is able to capture the overall molecular profile of small fragments and peptides, which are mainly formed during the duodenal phase, thus giving the kinetics of the whole digesti...
Food protein functionality: A comprehensive approach
Food Hydrocolloids, 2011
Food protein functionality has classically been viewed from the perspective of how single molecules or protein ingredients function in solutions and form simple colloidal structures. Based on this approach, tests on protein solutions are used to produce values for solubility, thermal stability, gelation, emulsifying, foaming, fat binding and water binding to name a few. While this approach is beneficial in understanding the properties of specific proteins and ingredients, it is somewhat restricted in predicting performance in real foods where the complexities of ingredients and processing operations have a significant affect on the colloidal structures and therefore overall properties of the final food product. In addition, focusing on proteins as just biopolymers used to create food structures ignores the biological functions of proteins in the diet. These can be beneficial, as in providing amino acids for protein synthesis or bioactive peptides, or these can be detrimental, as in causing a food allergic response. This review will focus on integrating the colloidal/polymer and biological aspects of protein functionality. This will be done using foams and gels to illustrate colloidal/polymer aspects and bioactive peptides and allergenicity to demonstrate biological function.
Relative Tryptic Digestion Rates of Food Proteins
Journal of Food Science, 1984
The kinetics of tryptic digestion of different food proteins were studied by measuring trichloroacetic acid soluble peptide release and peptide bond splitting either by proton titration at constant pH or by recording the pH drop in nonbuffered suspensions. The theoretical basis of the pH drop assay was described. Application of this method for comparative studies requires complementary determinations of buffering capacities of the samples and the time course of the reference protein digestion. It was shown that milk powder preparations differed in digestion rates by a factor of two and various lots of commercial soy meals differed as much as by a factor of three. Relative digestion rates of some food proteins from different sources could not be characterized by a single figure.
Food Chemistry
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