LAUSR

Graphical abstract Highlights • A protease from Mucor subtilissimus URM 4133, capable of producing bioactive peptides from goat casein, was purified.• Analysis by SDS-PAGE and zymography, shown a molecular mass of approximately 30 kDa.• Purified enzyme showed optimal activity and stability over a wide pH range (5–10), optimum temperature at 45–50 °C and stability above 80 % at 40 °C.• Activity was completely inhibited by PMSF indicating that it was a serine protease.• The kinetic and thermodynamic parameters of irreversible thermal denaturation, enthalpy, Gibbs free energy of denaturation were evaluated.• Peptide sequences compatible with this enzyme from M. subtilissimus URM 4133 were not found after analysis by MALDI-TOF suggesting a new protease.