Analyzing μ-Calpain induced proteolysis in a myofibril model system with vibrational and fluorescence spectroscopy.

Degree of post-mortem proteolysis influences overall meat quality (e.g. tenderness and water holding capacity). Degradation of isolated pork myofibril proteins by μ-Calpain for 0, 15 or 45 min was analyzed using four spectroscopic techniques; Raman, Fourier transform infrared (FT-IR), near infrared (NIR) and fluorescence spectroscopy. Sodium dodecyl sulfate polyacrylamide gel electrophoresis was used to determine degree of proteolysis. The main changes detected by FT-IR and Raman spectroscopy were degradation of protein backbones manifested in the spectra as an increase in terminal carboxylic acid vibrations, a decrease in CN vibration, as well as an increase in skeletal vibrations. A reduction in β-sheet secondary structures was also detected, while α-helix secondary structure seemed to stay relatively unchanged. NIR and fluorescence were not suited to analyze degree of proteolysis in this model system.