Browsing by Author "Maes E"

Now showing 1 - 4 of 4
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    Effect of Heat Treatment on Protein Self-Digestion in Ruminants' Milk
    (MDPI (Basel, Switzerland), 2023-09-21) Leite JAS; Montoya CA; Maes E; Hefer C; Cruz RAPA; Roy NC; McNabb WC; Liu Q; Liu H; Zhang J
    This study investigated whether heat treatments (raw, 63 °C for 30 min, and 85 °C for 5 min) affect protein hydrolysis by endogenous enzymes in the milk of ruminants (bovine, ovine, and caprine) using a self-digestion model. Self-digestion consisted of the incubation for six hours at 37 °C of the ruminants' milk. Free amino group concentration was measured by the o-phthaldialdehyde method, and peptide sequences were identified by chromatography-mass spectrometry. Results showed that heat treatments prior to self-digestion decreased the free NH2 by 59% in bovine milk heated at 85 °C/5 min, and by 44 and 53% in caprine milk heated at 63 °C/30 min and 85 °C/5 min, respectively. However, after self-digestion, only new free amino groups were observed for the raw and heated at 63 °C/30 min milk. β-Casein was the most cleaved protein in the raw and heated at 63 °C/30 min bovine milk. A similar trend was observed in raw ovine and caprine milk. Self-digestion increased 6.8-fold the potential antithrombin peptides in the bovine milk heated at 63 °C/30 min. Enhancing bioactive peptide abundance through self-digestion has potential applications in the industry for functional products. Overall, heat treatments affected the free amino groups according to the species and heat treatment applied, which was reflected in the varying degrees of cleaved peptide bonds and peptides released during self-digestion.
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    Effects of whey protein treatment in an in vitro intestinal cell model following oxidative stress or inflammatory challenge
    (Elsevier Ltd, 2025-01-28) Willems E; Purba A; Savoian MS; Hefer C; Maes E; Ulluwishewa D
    Bovine milk whey proteins with an isoelectric point >6.8 (‘whey’) have demonstrated anti-inflammatory and antioxidant properties. In the present study, pre-treatment of human intestinal cells (Caco-2) with whey mitigated intracellular reactive oxygen species produced in response to the pro-oxidant 2,2′-azobis (2-methylpropionamide)-dihydrochloride (AAPH). The mitigating effect was dose-dependent, and persisted when whey was removed prior to the addition of AAPH. Whey treatment also improved transepithelial electrical resistance (TEER), but returned to untreated-control levels upon removal of whey. Hence, whey can lead to cellular adaptations that aid intestinal function, but can exert additional properties while in contact with cells. Confocal imaging indicated that the previously observed TEER improvements in inflammatory-challenged Caco-2 monolayers were not due to the localisation of occludin or zonula occludens-1 tight junction proteins. However, proteomics analysis indicated a role for other tight junction proteins and provided insights into cellular adaptations that occur in response to whey pre-treatment.
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    Heat-Treatments Affect Protease Activities and Peptide Profiles of Ruminants' Milk
    (Frontiers Media S.A., 2021-03-10) Leite JAS; Montoya CA; Loveday SM; Maes E; Mullaney JA; McNabb WC; Roy NC; Abd El-Aty, AM
    Proteases present in milk are heat-sensitive, and their activities increase or decrease depending on the intensity of the thermal treatment applied. The thermal effects on the protease activity are well-known for bovine milk but poorly understood for ovine and caprine milk. This study aimed to determine the non-specific and specific protease activities in casein and whey fractions isolated from raw bovine, ovine, and caprine milk collected in early lactation, and to determine the effects of low-temperature, long-time (63°C for 30 min) and high-temperature, short-time (85°C for 5 min) treatments on protease activities within each milk fraction. The non-specific protease activities in raw and heat-treated milk samples were determined using the substrate azocasein. Plasmin (the main protease in milk) and plasminogen-derived activities were determined using the chromogenic substrate S-2251 (D-Val-Leu-Lys-pNA dihydrochloride). Peptides were characterized using high-resolution liquid chromatography coupled with tandem mass spectrometry. The activity of all native proteases, shown as non-specific proteases, was similar between raw bovine and caprine milk samples, but lower (P < 0.05) than raw ovine milk in the whey fraction. There was no difference (P > 0.05) between the non-specific protease activity of the casein fraction of raw bovine and caprine milk samples; both had higher activity than ovine milk. After 63°C/30 min, the non-specific protease activity decreased (44%; P > 0.05) for the bovine casein fraction only. In contrast, the protease activity of the milk heated at 85°C/5 min changed depending on the species and fraction. For instance, the activity decreased by 49% for ovine whey fraction, but it increased by 68% for ovine casein fraction. Plasmin and plasminogen were in general inactivated (P > 0.05) when all milk fractions were heated at 85°C/5 min. Most of the peptides present in heat-treated milk were derived from β-casein and αS1-casein, and they matched the hydrolysis profile of cathepsin D and plasmin. Identified peptides in ruminant milk samples had purported immunomodulatory and inhibitory functions. These findings indicate that the non-specific protease activity in whey and casein fractions differed between ruminant milk species, and specific thermal treatments could be used to retain better protease activity for all ruminant milk species.
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    Proteomic Profile of M. longissimus thoracis from Commercial Lambs Reared in Different Forage Systems
    (MDPI (Basel, Switzerland), 2022-05-13) Ye Y; Maes E; Deb-Choudhury S; Hefer CA; Schreurs NM; Realini CE; Colgrave M; Mora L
    This study compared the protein composition of M. longissimus thoracis of lambs from six commercial forage production systems in New Zealand. A total of 286 proteins were identified based on liquid chromatography-tandem mass spectrometry. First, a binomial model showed that different production groups could be distinguished based on abundances of 16 proteins. Second, pair-wise comparisons were performed to search for protein abundance differences in meat due to animal sex (ewe vs. wether), diet (perennial ryegrass vs. chicory), and age (4 vs. 6-8 months old). Greater abundance of some myofibrillar and sarcoplasmic proteins were observed in lamb loins from ewes compared to wethers. Chicory diet and older age at slaughter were associated with meat with lower abundance of some myofibrillar proteins, possibly due to a greater proportion of muscle glycolytic fibres. The proteins that showed significant differences in their abundances due to production factors could be further investigated to understand their influence on meat quality.