Thermodynamic and Enzymatic Study of Collagenolytic Protease Produced by Fermentation of Mucor subtilissimus UCP 1262

Collagenolytic proteases, known for their ability to degrade collagen, hold significance in various sectors, including biomedical, cosmetic, and food industries, due to the production of bioactive peptides. This study aims to partially characterize the collagenolytic protease from Mucor subtilissimus UCP 1262, highlighting its thermodynamic properties and potential industrial applications. The enzyme was produced via solid-state fermentation and purified through ion-exchange chromatography. Enzymatic activity was determined using specific substrates, demonstrating proteolytic activity of 98 U/mg and collagenolytic activity of 39.53 U/mg. Biochemical characterization revealed that the optimal temperature for enzyme activity is 40 °C, with an optimal pH of 7. Thermostability was assessed, showing that the enzyme retains 50% of its activity after 24 hours at 30-40 °C. The thermodynamic parameters of denaturation, such as Gibbs free energy (ΔG) and enthalpy (ΔH), were calculated, reaching ΔG up to 104.17 kJ/mol at 60ºC and ΔH of 29.12 kJ/mol, suggesting that the collagenolytic protease is promising for biomedical and industrial applications, particularly in processes involving collagen degradation.