Effect of enzymatic hydrolysis with Alcalase or Protamex on technological and antioxidant properties of whey protein hydrolysates.

The aim of this study was to evaluate the effect of the enzymatic hydrolysis, performed using Alcalase and Protamex enzymes, on the technological functionalities and the antioxidant capacity of whey protein hydrolysates (WPHs) to identify the conditions allowing to obtain target functionality/ies. Samples were characterized for hydrolysis degree (DH), molecular weight distribution, structural properties, and food-related functionalities. Free sulfhydryl groups and surface hydrophobicity significantly decreased with the increase in DH, regardless of the used enzyme. The foaming and antioxidant properties of Alcalase WPHs were higher as compared to those of WPI, reaching the maximum value at DH = 18-20 %, while higher DH resulted in impaired functionality. Gelling properties were guaranteed when WPI was hydrolysed by Protamex at DH < 15 % while foaming and antioxidant abilities were fostered at 15 < DH < 21 %. These results were well correlated with MW distribution and were rationalized into a road map which represents a useful tool in the selection of proper hydrolysis conditions (time, DH, enzyme type) to obtain WPHs with tailored functionalities. Research outcomes highlighted the possibility to drive protein hydrolysis to optimize the desired functionality/ies.

Unraveling the role of probiotics in affecting the structure of monoglyceride gelled emulsions: A low-field 1H NMR study.

The capacity of monoglyceride (MG) gelled emulsions (MEs) in protecting probiotic cells of Lacticaseibacillus rhamnosus against stresses suffered during food processing, storage, and human digestion has been recently demonstrated. These findings open new perspectives on the possible participation of probiotics in the stabilization of emulsion structure. To unravel this aspect, rheological analysis and Low-Field 1H NMR investigations were performed on MEs having different aqueous phases (water or skimmed milk) and stored for increasing time (1 and 14 days) at 4 °C. Loaded and unloaded samples were considered. Results highlighted that probiotics initially hindered the ability of MG to self-assemble in the multiphase environment, interacting in some way with MG crystalline lamellar structure, as confirmed by rheological and 1H NMR analysis. During storage, an increase of proton compartmentation was observed in loaded MEs indicating the role of probiotics in stabilizing MG structure at a molecular level. Such a result was more evident when the system was composed of milk, suggesting that the presence of milk-native components (i.e., lactose, proteins, and minerals) favored the cell-structure interactions. Such preliminary results could open new perspectives in considering probiotic cells as having an active role in the stabilization of food structure.

Design and advanced characterization of quercetin-loaded nano-liposomes prepared by high-pressure homogenization.

Quercetin-loaded nano-liposomes were prepared by high-pressure homogenization (HPH) at different pressures (up to 150 MPa) and number of passes (up to 3) to define the best processing conditions allowing the lowest particle size and the highest encapsulation efficiency (EE). The process at 150 MPa for 1 pass was the best, producing quercetin-loaded liposomes with the lowest particle size and 42% EE. Advanced techniques (multi-detector asymmetrical-flow field flow fractionation and analytical ultracentrifugation combined with transmission electron microscopy) were further used for the characterization of the liposomes which were oblong in shape (ca. 30 nm). Results highlight the need for several techniques to study nano-sized, polydisperse samples. The potential of quercetin-loaded liposomes against colon cancer cells was demonstrated. Results prove that HPH is an efficient and sustainable method for liposome preparation and highlight the remarkable role of process optimisation as well as the powerfulness of advanced methodologies for the characterisation of nano-structures.

Adult and elderly in vitro digestibility patterns of proteins and carbohydrates as affected by different commercial bread types.

Carbohydrate and protein digestibility were assessed in different commercial bread types, i.e., soft, durum, and whole wheat, by applying in vitro digestion protocols mimicking adult or elderly physiological conditions. Protein digestibility was measured after the gastric and intestinal phases by the o-phthalaldehyde spectrophotometric assay (OPA). Carbohydrate digestibility was assessed by determining the incremental area under glucose curve during the intestinal phase of digestion to estimate the glycaemic index (GIe). Finally, the correlation between protein and carbohydrate digestibility was computed. Bread proteins presented a high gastric resistance, with a digestibility < 10% in all cases while after the intestinal phase, protein digestibility increased, ranging from 40 to 70%. Protein digestibility was affected both by formulation, with whole wheat bread presenting the lowest values, and by physiological conditions, with restrained digestibility under elderly conditions compared to adult ones. The GIe decreased in the order durum > soft > whole, under both adult (119, 101, and 82, respectively) and elderly (107, 93, and 65, respectively) conditions. The extent of differences under different physiological settings varied depending on the bread type, without significant changes for soft wheat bread (ΔGIe = 8), whereas elderly conditions significantly reduced GIe both for durum (ΔGIe = 12) and whole (ΔGIe = 17) wheat bread. An almost strong positive correlation between protein digestibility and GIe was observed (p = 0.69), indicating that concomitantly addressing sarcopenia and type 2 diabetes can only be pursued by a compromise solution or more desirably requires identifying technological strategies to maximize protein digestibility while restraining the glycaemic response.

Comparison of protein in vitro digestibility under adult and elderly conditions: The case study of wheat, pea, rice, and whey proteins.

In this study an in vitro static digestion method mimicking the elderly gastrointestinal conditions was designed by adapting the physiological parameters described in the INFOGEST standardized static in vitro digestion protocol, i.e., pH, digestive phase duration, concentrations of enzymes and bile salts, to the aged GI transit. The digestibility of proteins from different sources (pea, rice, wheat, and milk whey) was then assessed. Protein digestive behaviour was monitored after gastric and intestinal phases by BCA assay and SDS-PAGE to assess protein hydrolysis both from a quantitative and a qualitative point of view. Digested samples were also analysed for physical characteristics in terms of particle size and zeta potential. Data acquired under elderly gastrointestinal conditions were compared to those obtained by using the INFOGEST protocol designed to study adult digestion. Results clearly showed that the elderly gastrointestinal conditions deeply affected proteolysis leading to a general reduction of protein digestibility in comparison to the adult model. The proteolysis extent depended on the protein source with whey and rice proteins showing about 20% reduction using the model mimicking the elderly gut, followed by pea (about 10% reduction) and wheat (about 4% reduction) proteins. The knowledge of protein digestibility under elderly gastrointestinal conditions generated in this study could be useful in the attempt to develop age-tailored products.

Steering protein and lipid digestibility by oleogelation with protein aerogels.

The aim of the present work was to assess the effect of an innovative oleogelation strategy, the aerogel-template approach, on protein and lipid digestibility. Whey protein isolate (WP) was converted into aerogel particles via supercritical CO2 drying. Oleogels were then prepared by absorption of sunflower (SO) or flaxseed (FLX) oil (80%, w/w) into the aerogel particle template and subjected to in vitro digestion. WP aerogel-templated oleogels showed a specific destructuring behaviour during digestion. Confocal micrographs clearly demonstrated that the original oleogel structure was lost at the gastric level, with the release of oil droplets smaller (D32 < 10 µm) than those observed in the case of the unstructured oils (D32 > 30 µm), stabilised by undigested aerogel proteins. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and bicinchoninic acid (BCA) assay confirmed that aerogelation reduced the gastric proteolysis of WP from nearly 100% to 70%. The digestion of the SO oleogel led to similar gastric protein digestibility. In contrast, in the case of the FLX oleogel, gastric proteolysis decreased to 40%, suggesting a role of the oil nature in steering WP aerogel digestion. In all cases, upon intestinal digestion aerogel proteins resulted completely hydrolysed. The lipolysis degree of SO (75%) and FLX (34%) oil in the oleogels was higher than that of the unstructured SO (66%) and FLX (24%) oils, due to the larger surface offered by smaller oil droplets to the action of intestinal lipases. This was confirmed by dynamic light scattering, showing a shift towards smaller size in the digestive micelle distribution of oleogels at the end of the intestinal phase. Oleogelation through the WP aerogel-template approach could be regarded as a strategy to steer lipid digestibility while also modulating the release of bioaccessible peptides.

Designing food for the elderly: the critical impact of food structure.

Ageing is an unavoidable progressive process causing many changes of the individual life. However, if faced in an efficient way, living longer in a healthy status could be an opportunity for all. In this context, food consumption and dietary patterns are pivotal factors in promoting active and healthy ageing. The development of food products tailored for the specific needs of the elderly might favour the fulfilment of nutritionally balanced diets, while reducing the consequences of malnutrition. To this aim, the application of a food structure design approach could be particularly profitable, being food structure responsible to the final functionalities of food products. In this narrative review, the physiological changes associated to food consumption occurring during ageing were firstly discussed. Then, the focus shifted to the possible role of food structure in delivering target functionalities, considering food acceptability, digestion of the nutrients, bioactive molecules and probiotic bacteria.

Correction: Effect of the formulation and structure of monoglyceride-based gels on the viability of probiotic Lactobacillus rhamnosus upon in vitro digestion.

Correction for 'Effect of the formulation and structure of monoglyceride-based gels on the viability of probiotic Lactobacillus rhamnosus upon in vitro digestion' by Sofia Melchior et al., Food Funct., 2021, DOI: 10.1039/D0FO01788D.

Effect of the formulation and structure of monoglyceride-based gels on the viability of probiotic Lactobacillus rhamnosus upon in vitro digestion.

This research was conducted to evaluate the potential use of saturated monoglyceride (MG)-based gels in the protection of probiotics upon in vitro digestion. For this purpose, a Lactobacillus rhamnosus strain was inoculated into binary and ternary systems, containing MGs, a water phase composed of an aqueous solution at controlled pH or UHT skimmed milk, and in ternary gels, sunflower oil. Gel structure characterization was initially performed just after preparation and after 14 days of storage at 4 °C by rheological, mechanical, thermal, and microscopy analyses. Afterwards, probiotic viability upon in vitro digestion was evaluated. The results highlighted that all freshly prepared samples showed good capability to protect L. rhamnosus with the exception of the binary system containing milk. However, the digestion of samples after 14 days of storage showed that the ternary system containing skimmed milk exhibited the best protection performance ensuring a L. rhamnosus viability of almost 106 CFU g-1 at the end of the gastrointestinal passage. Confocal microscopy results demonstrated that bacterial cells were located prevalently within the aqueous domain near the monoglycerides and protein aggregates. Under these conditions, they can simultaneously achieve physical protection and find nutrients to survive environmental stresses. These findings suggest that MG-based gels can be proposed as efficient carriers of probiotic bacteria not only during food processing and storage but also upon digestion.

Application of high-pressure homogenization to tailor the functionalities of native wheat starch.

BACKGROUND: The effect of high-pressure homogenization (HPH) on the rheological and thermal properties, water retention capacity (WRC), morphology and in vitro digestion of wheat starch was evaluated. Starch suspensions (50 g kg-1 , w/w) were treated at increasing pressures (up to 100 MPa) and numbers of cycles (up to 5) to generate a wide range of energy densities (70-500 MJ m-3 ) delivered to the sample during processing. RESULTS: High-pressure homogenization induced a partial starch gelatinization confirmed by higher digestibility. Gelatinization degree (GD) was between 13% and 83%, causing a wide range of functional properties. High-pressure homogenization-treated starch samples showed WRC values of 810-1910 g kg-1 . Storage modulus (G') and complex viscosity (η* ) of starch dispersions were almost two and three times higher than the control at 13% and 83% GD, respectively. Positive linear relationships between GD (R = 0.98, P < 0.001), WRC (R = 0.87, P < 0.05), or rheological parameters (R = 0.89÷0.90, P < 0.01) and energy density of HPH treatments were found. CONCLUSION: High-pressure homogenization treatment represents a promising technology to obtain wheat starch with tailored rheological properties and digestibility, which allows the texture and glycemic response of food products to be adjusted. © 2020 Society of Chemical Industry.

Effect of different biopolymer-based structured systems on the survival of probiotic strains during storage and in vitro digestion.

BACKGROUND: This study aimed to evaluate the protective effect of different biopolymer systems on the viability of two probiotics (Lactobacillus rhamnosus and Streptococcus thermophilus) during storage and in vitro digestion. Methylcellulose (MC), sodium alginate (SA), and whey protein (WP)-based structures were designed and characterized in terms of pH, rheological properties, and visual appearance. RESULTS: The results highlighted that the WP-system ensured probiotic protection during both storage and in vitro digestion. This result was attributed to a combined effect of the physical barrier offered by the protein gel network and whey proteins as a nutrient for microbes. On the other hand, surprisingly, the viscous methylcellulose-based system was able to guarantee good microbial viability during storage. However, this was not confirmed during in vitro digestion. The opposite results were obtained for sodium alginate beads. CONCLUSION: The results suggest that the capacity of a polymeric structure to protect probiotic bacteria is a combination of structural organization and system formulation. © 2020 Society of Chemical Industry.