Revisiting ultrasound and high-pressure homogenization modification of pea protein isolate: insights into gut...

Ultrasound (US) and high-pressure homogenization (HPH) have been reported to enhance the techno-functionality of pea protein isolate (PPI); however, their impacts on the nutritional attributes of PPI remain unclear. This study investigated whether US- and HPH-induced structural modifications of PPI could influence its digestibility, gut microbiota modulation, and anti-inflammatory activity. PPI was treated with US (40% and 80% amplitude) and HPH (120 and 240 MPa), followed by digestion using the INFOGEST model. All modified PPIs exhibited significantly improved in vitro digestibility. In vitro colonic fermentation showed that PPI shaped a distinct gut microbial profile compared with the blank. However, US and HPH treatments did not further alter microbial diversity or community structure. LEfSe analysis revealed consistent genus-level shifts among all PPIs. In RAW 264.7 macrophage model, unmodified, US- and HPH-treated PPIs exhibited similar anti-inflammatory effects, reducing NO and pro-inflammatory cytokines at higher concentrations. Overall, despite substantially altering the structure and digestibility of PPI, US and HPH did not influence pea protein's gut microbiota-modulating or anti-inflammatory properties. These findings suggest that US and HPH can be safely applied to enhance PPI functionality without compromising its nutritional relevance.