Inactivation kinetics and secondary structural change of PEF-treated POD and PPO

Abstract

Effects of pulsed electric fields (PEF) on the activity of peroxidase (POD) and polyphenol oxidase (PPO) in buffered solution were studied while the corresponding changes to their secondary structures was demonstrated by far-UV Circular dichroism (CD). The relative residual activity of POD and PPO decreased with the increase in electric field strength and treatment time, and PPO was more susceptible than POD to PEF treatment. The greatest reduction of the activity was achieved for POD at 25 kV/cm for 1740 μs and PPO at 25 kV/cm for 744 μs with reductions of 32.2% and 76.2%, respectively. The inactivation kinetic parameters D-value and Z E value were calculated. The D-values of PPO were smaller than those POD at higher electric field strength, and Z E values of POD and PPO were 36.9 and 16.2 kV/cm, respectively. The secondary structures of the two enzymes were changed following treatment by PEF. The intensity of negative peaks in the CD spectra decreased, and the CD spectra of PPO changed more significantly than that of POD; the reduction of the relative α-helix fractions for POD at 25 kV/cm for 124 μs was 22.63% while it was 50.72% for PPO at 25 kV/cm for 52 μs. The inactivation of PEF-treated POD and PPO was in close agreement with their secondary structure changes.