Delivery of Phenolic Compounds, Peptides and β-Glucan to the Gastrointestinal Tract by Incorporating Dietary Fibre-Rich Mushrooms into Sorghum Biscuits

Sorghum biscuits were enriched with mushroom powders (Lentinula edodes, Auricularia auricula and Tremella fuciformis) at 5%, 10% and 15% substitution levels. An in vitro gastrointestinal digestion was used to evaluate the effect of this enrichment on the phenolic content and soluble peptide content as well as antioxidant activities of the gastric or intestinal supernatants (bio-accessible fractions), and the remaining portions of phenolic compounds, antioxidants and β-glucan in the undigested residue (non-digestible fraction). The phenolic content of the gastric and intestinal supernatants obtained from digested mushroom-enriched biscuits was found to be higher than that of control biscuit, and the phenolic content was positively correlated to the antioxidant activities in each fraction (p < 0.001). L. edodes and T. fuciformis enrichment increased the soluble protein content (small peptide) of sorghum biscuits after in vitro digestion. All mushroom enrichment increased the total phenolic content and β-glucan content of the undigested residue and they were positively correlated (p < 0.001). The insoluble dietary fibre of biscuits was positively correlated with β-glucan content (p < 0.001) of undigested residue. These findings suggested that enriching food with mushroom derived dietary fibre increases the bioavailability of the non-digestible β-glucan and phenolic compounds.

Changes in flavor compounds of Aloididae aloidi during enzymatic hydrolysis

Protamex was selected to prepare the hydrolysate. E-tongue, free amino acid combined with soluble peptide analysis were used to detect the flavor changes of Aloididae aloidi during enzymolysis. Degree of proteolysis increased with the prolongation of enzymolysis time, and reached the maximum value at 8 hours. The content of soluble peptide of hydrolysate increased firstly and then decreased in the later process. The E-tongue could effectively distinguish the taste difference of hydrolysates at different enzymolysis time, and the hydrolysate presented strong bitterness and astringency during the whole enzymolysis. The total amount of free amino acids in the hydrolysate increased gradually, and some sweet, umami and bitter amino acids increased in varying degrees during the process of enzymolysis.

Optimization of Liposomes for Antigen Targeting to Splenic CD169+ Macrophages

Despite promising progress in cancer vaccination, therapeutic effectiveness is often insufficient. Cancer vaccine effectiveness could be enhanced by targeting vaccine antigens to antigen-presenting cells, thereby increasing T-cell activation. CD169-expressing splenic macrophages efficiently capture particulate antigens from the blood and transfer these antigens to dendritic cells for the activation of CD8+ T cells. In this study, we incorporated a physiological ligand for CD169, the ganglioside GM3, into liposomes to enhance liposome uptake by CD169+ macrophages. We assessed how variation in the amount of GM3, surface-attached PEG and liposomal size affected the binding to, and uptake by, CD169+ macrophages in vitro and in vivo. As a proof of concept, we prepared GM3-targeted liposomes containing a long synthetic ovalbumin peptide and tested the capacity of these liposomes to induce CD8+ and CD4+ T-cell responses compared to control liposomes or soluble peptide. The data indicate that the delivery of liposomes to splenic CD169+ macrophages can be optimized by the selection of liposomal constituents and liposomal size. Moreover, optimized GM3-mediated liposomal targeting to CD169+ macrophages induces potent immune responses and therefore presents as an interesting delivery strategy for cancer vaccination.

Peptide Profiling and Biological Activities of 12-Month Ripened Parmigiano Reggiano Cheese

Proteolysis degree, biological activities, and water-soluble peptide patterns were evaluated in 12 month-ripened Parmigiano Reggiano (PR) cheeses collected in different dairy farms and showing different salt and fat content. Samples classified in high-salt and high-fat group (HH) generally showed lower proteolysis degree than samples having low-salt and low-fat content (LL). This positive correlation between salt/fat reduction and proteolysis was also confirmed by the analysis of biological activities, as the LL group showed higher average values of angiotensin-converting enzyme (ACE)-inhibitory and antioxidant activities. UHPLC/HR-MS allowed the identification of 805 unique peptides: LL and HH groups shared 59.3% of these peptides, while 20.9% and 19.9% were LL and HH specific, respectively. Frequency analysis of peptides identified a core of 183 peptides typical of 12-month ripened PR cheeses (corresponding to the 22.7% of total peptides), but no significant differences were detected in peptide patterns between LL and HH groups. Forty bioactive peptides, including 18 ACE-inhibitors and 12 anti-microbial peptides, were identified, of which 25 firstly found in PR cheese. Globally, this work contributed to unraveling the potentially healthy benefits of peptides fraction in PR cheese and provided prior evidence that PR with reduced fat/salt content showed the highest antihypertensive and antioxidant activities.

Functional Properties of Defatted Roselle (Hibiscus sabdariffa L.) Seeds Protein and Its Hydrolysates

Aim: The purpose of this study was to evaluate the functional properties of Roselle seed protein isolates and its hydrolysates. Place and Duration of Study: The Roselle seeds were collected in Koutiala (Mali), in November 2018. All analysis were conducted in the Faculty of Sciences and Technics, particularly in the Laboratory of Plant and Food Biochemistry and Biotechnology from January to June 2019. Methodology: The effect of enzymatic hydrolysis on the functional properties of Roselle seed protein (RSP) was investigated. Defatted Roselle seed flour was used to extract the protein isolates. The protein was digested for 2 hours and 3 hours using pepsin followed by pancreatin. Results: The 2 hours and 3 hours Roselle seed protein hydrolysates (RSPH2, RSPH3) compare to RSPI, exhibited a good foaming capacity of 300, 315 and 165% respectively. The water holding capacity (WHC) of the RSPI, RSPH2 and the RSPH3 were 2, 2.5 and 2.2 ml/g respectively. The oil holding capacity ranged from 5.75 to 5.32 ml/g, the emulsifying capacity of the RSPH2 was higher than that of the RSPH3 and the RSPI, 105, 97 and 82 ml/g respectively. Conclusion: The ability of pepsin and pancreatin hydrolysates to be functional is primarily due to their soluble peptide content. The samples have good functional properties. These results proposed that pepsin and pancreatin hydrolysates could be useful as whole or partial replacement of high-price materials such as egg albumen and casein.

Production of soluble pMHC-I molecules in mammalian cells using the molecular chaperone TAPBPR

Current approaches for generating major histocompatibility complex (MHC) Class-I proteins with desired bound peptides (pMHC-I) for research, diagnostic and therapeutic applications are limited by the inherent instability of empty MHC-I molecules. Using the properties of the chaperone TAP-binding protein related (TAPBPR), we have developed a robust method to produce soluble, peptide-receptive MHC-I molecules in Chinese Hamster Ovary cells at high yield, completely bypassing the requirement for laborious refolding from inclusion bodies expressed in E.coli. Purified MHC-I/TAPBPR complexes can be prepared for multiple human allotypes, and exhibit complex glycan modifications at the conserved Asn 86 residue. As a proof of concept, we demonstrate both HLA allele-specific peptide binding and MHC-restricted antigen recognition by T cells for two relevant tumor-associated antigens. Our system provides a facile, high-throughput approach for generating pMHC-I antigens to probe and expand TCR specificities present in polyclonal T cell repertoires.