A Rapid and Efficient Method for Purification of Peroxidase from Opuntia-ficus indica Stem with Decolorization Effect
Peroxidase enzyme was purified from Opuntia ficusindica stem for decolorization some synthetic dyes including congo red, methyl red, indigo carmine, crystal violet and ponceau red by TPP. For the purification of the enzyme, the stem of a fresh plant was first homogenized by using phosphate buffer, and subsequently, supernatant was filtered. The homogenate was mixed with ammonium sulfate at room temperature, and t-butanol was added. The intermediate phase was separated and optimized according to different process parameters as homogenate/t-butanol ratio (1:0.5), ammonium sulfate concentration (40%), pH (8), temperature (30°C) and values of KM (4.8 mM) and Vmax (0,005 U/mL.min) with 2,25-fold purification and 106,64% recovery. The molecular weight of partitioned enzyme was found about 28 kDa compared with protein standard. TPP was found to be an easy and effective applicable technique with utilization for extraction and purification of peroxidase enzyme and the purified enzyme decolorized the tested synthetic dyes, successfully. The most important advantage of the method is a high recovery of peroxidase enzyme without requirement any column.
