Effect of ellagitannins gut microbiota metabolites ellagic acid (EA) and urolithin A-urolithin D (UroA-UroD) on human serum albumin (HSA) glycation were firstly evaluated in this research. The inhibition mechanisms were investigated by methylglyoxal (MGO) trapping and radical scavenging ability assays, docking studies and Nano LC-Orbitrap-MS/MS technology. Results indicated that the inhibition of urolithins on HSA glycation was highly positive correlated with the number of phenolic hydroxy groups. Addition of UroD and EA could effectively enhance the content of free amino group, suppress dicarbonyl compounds and advanced glycation end-products (AGEs) formation, alleviated tryptophan and protein oxidation, inhibited HSA amyloid-like aggregation. They could also trap MGO and scavenge DPPH· and ABTS·⁺. Molecular docking indicated that EA and UroA-UroD interact with HSA mainly through hydrogen bound and hydrophobic interaction, among which two or three hydrogen bonds were formed. The number of glycation sites were reduced from 11 to 10, 10, 7, and 10, respectively, when 90 μmol/L of EA, UroA, UroC and UroD were added. However, weak inhibition was observed on UroA and UroB. These findings can provide scientific evidence for the application of ellagitannins-rich foods in alleviating diabetic complications.

Gut microbiota plays an important role in food allergy. The immunoglobulin G (IgG)/ immunoglobulin E (IgE) binding capacity and human gut microbiota changes of digestion products derived from glycated ovalbumin (OVA) were investigated. Gastrointestinal digestion effectively destroyed the primary structure of glycated OVA, resulting in a significantly higher digestibility than gastric digestion, and more abundant peptides below 3 kDa. Moreover, gastric and gastrointestinal digestion products have different fluorescence quenching and red shift of fluorescence peaks, and possess different conformational structures. These changes resulted in a decrease in 28.7% of the IgE binding capacity of gastrointestinal digestion products beyond that of pepsin. Moreover, gastrointestinal digestion products of glycated OVA increased significantly the proportion of Subdoligranulum, Collinsella, and Bifidobacterium. Therefore, gastrointestinal digestion products of glycated OVA altered human intestinal microbiota, reducing the risk of potential allergy.

Bovine α-lactalbumin (BLA) induced severe cow's milk allergy. In this study, a novel strategy combining ultrasonication, performed before glycation, and phosphorylation was proposed to reduce BLA allergenicity. Result showed that IgE- and IgG-binding capacities and the release rates of histamine and interleukin-6 from RBL-2H3 were reduced. Moreover, intrinsic fluorescence intensity and surface hydrophobicity were decreased, whereas glycated sites (R10, N44, K79, K108, N102 and K114) and phosphorylated sites (Y36 and S112) of BLA were increased. Minimum allergenicity was detected during BLA treatment after ultrasonic prior to glycation and subsequent phosphorylation because of considerable increase in glycated and phosphorylated sites. Therefore, the decrease in allergenicity of BLA, the effect correlated well with the shielding effect of glycated sites combined with phosphorylated sites and the conformational changes. This study provides important theoretical foundations for improving and using the ultrasonic technology combined with protein modification in allergenic protein processing.