Continuous production of velvet bean-based bioactive peptides in membrane reactor with dual enzyme system.

• The optimum conditions were at [E]/[S] = 5%, pH = 7.5, and τ = 12 h. • The long-term continuous production was performed for 72 h. • Membrane fouling mitigations: threshold flux and membrane cleaning. • IC 50 values of 17.85 and 4.58 µg protein/mL for antioxidant and ACE inhibition. • DPP-4 and ACE inhibitor as the "theoretical" predominant activities. One of the main challenges hindering the commercialization of bioactive peptides is the lack of scalable and consistent production methods. To overcome this obstacle, an automated enzyme membrane reactor was used to continuously produce bioactive peptides from velvet bean (Mucuna pruriens). The optimum operating conditions were [E]/[S] = 5%, pH = 7.5, and τ = 12 h. The long-term continuous operation of the EMR system demonstrated its ability to maintain steady-state conditions. To minimize membrane fouling, an industrially viable strategy was employed, which combines operation at threshold flux and performing regular membrane cleaning. Further fractionation of the hydrolysates with a 2-kDa PES membrane resulted in the highest bioactivity. The IC 50 values for antioxidant and ACE inhibition were 17.85 and 4.58 µg protein/mL, respectively. To map the overall bioactivities of the hydrolysates, LC-MS analysis coupled with BIOPEP-UWM database was performed and obtained DPP-4 and ACE inhibitors as the predominant bioactive activities. [ABSTRACT FROM AUTHOR]