Simulated Gastrointestinal Digestion of Cocoa: Detection of Resistant Peptides and In Silico/In Vitro Prediction of Their Ace Inhibitory Activity

Abstract

In this study we investigated the oligopeptide pattern in fermented cocoa beans and derived products after simulated gastrointestinal digestion. Peptides in digested cocoa samples were identified based on the mass fragmentation and on the software analysis of vicilin and 21 KDa cocoa seed protein sequences, the most abundant cocoa proteins. Quantification was carried out by liquid chromatography/electrospray ionisation mass spectrometry (LC/ESI-MS) using an internal standard. Sixty five peptides were identified in the digested samples, including three pyroglutamyl derivatives. The in vitro angiotensin-converting enzyme (ACE)-inhibitory activity of cocoa digests were tested, demonstrating a high inhibition activity, especially for digestates of cocoa beans. The peptides identified were screened for their potential ACE inhibitory activity through an in silico approach, and about 20 di-, three- and tetra-peptides actually present in our samples were predicted as active. Two of the potentially active peptides were chemically synthesized and then assessed for their inhibitory activity by using the ACE in vitro assay. These peptides demonstrated an ACE inhibitory activity, however, that was too weak to explain alone the high activity of cocoa digestates, suggesting a synergic effect of all cocoa peptides. As a whole, results showed that an average chocolate portion (30 g) ensures an amount of peptides after digestion that, assuming complete absorption, could reach almost a complete inhibition of ACE.

Study of Polyphenol Content and Antioxidant Properties of Various Mix of Chocolate Milk Masses with Different Protein Content

“…The peptides come from two main protein fractions-globules, consisting of a storage protein resembling vicilin and albumin, which exhibit trypsin inhibitory properties. Cocoa proteins during natural cocoa fermentation are split into hydrophilic and hydrophobic peptides as well as amino acids by autolysis of two endogenous enzymes: aspartic endoprotease and carboxypeptidase activated by microbial metabolites such as acetic acid [38]. Milk proteins include complex groups of proteins: caseins (αs1-, αs2-, β-, and κcasein), whey proteins (β-lactoglobulin, α-lacto-albumin, albumin serum, and immunoglobulin), and protein envelopes of fat globules.…”

Impacts of cocoa pod maturity at harvest and bean fermentation period on the production of chocolate with potential health benefits

“…Pod harvest maturity and post-harvest processes such as fermentation and roasting not only contribute to the formation of flavor and aroma in beans but also determine the development and production of bioactive compounds with health benefits. 13,14 Peptides and amino acids developed during fermentation are important precursors to trigger the development of the cacao aroma. 15,16 Cacao pulp, which is rich in sugars, pectin, and citric acid, is an important substrate for microorganisms (yeasts, lactic acid bacteria, and acetic acid bacteria) during fermentation.…”

“…Pod harvest maturity and post‐harvest processes such as fermentation and roasting not only contribute to the formation of flavor and aroma in beans but also determine the development and production of bioactive compounds with health benefits 13,14 . Peptides and amino acids developed during fermentation are important precursors to trigger the development of the cacao aroma 15,16 .…”

Effects of Raw and Roasted Cocoa Bean Extracts Supplementation on Intestinal Enzyme Activity, Biochemical Parameters, and Antioxidant Status in Rats Fed a High-Fat Diet

“…Although ancient civilizations recognized cocoa beans as a health-promoting raw material, cocoa-derived products have only recently gained recognition for use as a significant source of functional ingredients in foods, nutraceuticals, or cosmetics [4]. These beneficial properties, attributed mainly to the presence of a significant number of bioactive compounds, are a perennial topic of investigation [1,5,6]. Cocoa beans due to the presence of polyphenols exhibit diverse biological activities, such as antioxidant, antiradical, antimicrobial, anti-inflammatory, antithrombotic, antihypertensive, anticarcinogenic, antiallergic, immunomodulatory, and cardioprotective, resulting in the protection against diseases such as cardiovascular diseases, cancer or neurodegenerative disorders among others [7][8][9][10][11][12][13][14][15].…”