Pulsed electric field pretreatment of bovine α-lactalbumin: Effect of processing parameters on structure, hydrolysis and production of potential antihypertensive and antidiabetic peptides

α-lactalbumin (α-lac), a key protein in the agro-food industry, is a promising source of bioactive peptides with antihypertensive and antidiabetic potential. However, its compact globular structure limits enzymatic hydrolysis efficiency. This study demonstrates the innovative use of pulsed electric fields (PEF) as a pretreatment to enhance hydrolysis compared to untreated and thermally pretreated samples. PEF conditions of 10 and 15 kV/cm (75 pulses) doubled the degree of hydrolysis (DH), while 20 kV/cm (37 pulses) achieved a 1.8-fold increase relative to native α-lac. Structural analyses revealed that PEF induced partial protein unfolding, increasing the accessibility of cleavage sites. Furthermore, PEF pretreatment facilitated the generation of bioactive peptides such as AHKAL and LAHK, which have not been previously reported for their antihypertensive and antidiabetic activities. These peptides showed an average abundance 2 times higher than in native α-lac and exceeded yields from thermal pretreatments. PEF also proved to be eco-efficient, with conditions comparable to thermal treatment at 75 °C and without excessive denaturation. This study establishes PEF as an innovative and sustainable technology capable of unlocking the functional potential of α-lac, paving the way for new opportunities in protein valorization within the agro-food industry.