BACKGROUND: Bioactive peptides derived from milk proteins are recognized as functional foods, but their consumption is limited by undesirable or bitter flavour, poor stability, and low bioavailability. Electrospinning is a versatile process for encapsulation of various bioactive compounds in the form of nanosized fibres, which can circumvent these disadvantages. This study was aimed at the preparation of casein-derived peptides-loaded nanofibres through electrospinning and characterizing them for fortification of milk.RESULTS: Pullulan at 100, 120, and 140 g kg −1 concentrations was used for electrospinning of peptides. Scanning electron and atomic force micrographs revealed the formation of clean bead-free peptides-loaded pullulan nanofibres at 120 and 140 g kg −1 concentrations with mean diameter of 60.45-133.05 nm and encapsulation efficiency of 72.95-86.04%. Fourier transform infrared spectra and X-ray diffractograms revealed the absence of interactions between the functional groups of pullulan and peptides during electrospinning. The zeta potential of the peptides-loaded nanofibres ranged from −15.6 to −24.6 mV, and the hydrodynamic diameter varied from 118.7 to 256.2 nm. The peptides from electrospun nanofibres showed sustained release to the extent of 75.3% after 8 h in gastrointestinal pH conditions. The release kinetics of peptides from nanofibres was best fitted to a Peppas-Sahlin model (R 2 = 0.987), and through diffusion and erosion mechanisms. The antioxidant activity of pure peptides and those from nanofibres was comparable. The physico-chemical qualities of milk fortified with encapsulated peptides did not show noticeable difference either. CONCLUSIONS: From the morphological, ultrastructural, particle size, encapsulation efficiency, release kinetics, and antioxidant activity data, it was inferred that electrospinning could be an effective technique for nanoencapsulation of casein-derived bioactive peptides. These peptides-loaded nanofibres could be used for fortification of milk and milk products.