Compressed propane as a new and fast method of pre-purification of radish (Raphanus sativus L.) peroxidase

The present study describes a simple protocol to purify Radish (Raphanus sativus L.) peroxidase (RP) and, consequently, shows the enhancement of its specific activity after treatment with compressed propane. In addition, the evaluation of temperature (30–50 °C) and pressure (50–200 bar) effect on peroxidase activity and stability in dense propane medium is also shown. In each experimental condition, enzyme extract samples were analyzed after 5, 15, 30, 60, and 120 min of high pressure treatment. Peroxidase remained active (U/mL) after 120 min incubation at 30 and 40 °C, while at 50 °C, the enzyme activity (U/mL) was lower at 200 bar (residual activity of 50%) than at 50 bar (67% residual activity). Lyophilized sample treatment led to an increase in RP activity (115% residual activity). This result indicates that the treatment with compressed propane is a viable technique for extract purification as well as specific activity increase of peroxidases.