Hydrolysis of pea protein differentially modulates its effect on iron bioaccessibility, sulfur availability, composition and activity of gut microbial communities in vitro

Both plant proteins and iron supplements can demonstrate high susceptibility to escape small intestinal digestion and absorption, hence are often present throughout colonic fermentation. Whilst colonic iron delivery may adversely affect the gut microbiota and epithelial integrity, nascent evidence suggests that pea proteins may possess beneficial prebiotic and antioxidant effects during gut fermentation. This study investigated the interaction between exogenously added iron and pea protein isolate (PPI) or pea protein hydrolysate (PPH) during in vitro gastrointestinal digestion and colonic fermentation. Results revealed that enzymatic hydrolysis mitigated the crude protein's inhibitory effects on iron solubility during small intestinal digestion. Colonic fermentation of iron-containing treatments led to an increase in iron bioaccessibility and was characterized by a loss of within-species diversity, a marked increase in members of Proteobacteria, and eradication of some species of Lactobacillaceae. Although these patterns were also observed with pea proteins, the extent of the effects differed. Only PPI displayed significantly higher levels of total short-chain fatty acids in the presence of iron, accompanied by greater abundance of Propionibacteriaceae relative to other treatments. Additionally, we provide evidence that the iron-induced changes in the gut microbiome may be associated with its effect on endogenous sulfur solubility. These findings highlight the potential trade-off between protein-induced enhancements in fortified iron bioaccessibility and effects on the gut microbiome, and the role of iron in facilitating colonic sulfur delivery.