BackgroundIndustrial processing can alter the structural complexity of dietary proteins and, potentially, their digestion and absorption upon ingestion. High moisture extrusion (HME), a common processing method used to produce meat alternative products, affects in vitro digestion, but human data are lacking. We hypothesised that HME of a mycoprotein/pea protein blend would impair in vitro digestion and in vivo postprandial plasma amino acid availability. MethodsIn Study A nine healthy volunteers completed two experimental trials in a randomised, double-blind, cross-over design. Participants consumed a beverage containing 25 g protein from a ‘dry’ blend (CON) of mycoprotein/pea protein (39/61%) or a HME content matched blend (EXT). Arterialised-venous blood samples were collected in the postabsorptive state and regularly over a 5 h postprandial period to assess plasma amino acid concentrations. In Study B, in vitro digestibility of the two beverages were assessed using BCA assay and optical-fluorescence microscopy at baseline, during and following gastric and intestinal digestion using the INFOGEST model of digestion. ResultsProtein ingestion increased plasma total, essential (EAA), and branched-chain amino acid (BCAA) concentrations (time effect; P<0.0001), but more rapidly and to a greater magnitude in the CON compared with the EXT condition (condition x time interaction; P<0.0001). This resulted in greater plasma availability of EAA and BCAA concentrations during the early postprandial period (0-150 min). These data were corroborated by the in vitro approach which showed greater protein availability in the CON (2150 ± 129 mg·mL-1) compared with EXT (590 ± 41 mg·mL-1) during the gastric phase. Fluorescence microscopy revealed clear structural differences between the two conditions. ConclusionsThese data demonstrate that HME delays in vivo plasma amino acid availability following ingestion of a mycoprotein/pea protein blend. This is likely due to impaired gastric phase digestion as a result of HME induced aggregate formation in the pea protein. Clinical trialsNCT05584358