Abstract
This paper reports the isolation and purification of peroxidase from low cost material; moreover, no significant work has been done on the isolation and purification of peroxidase from such cost effective sources (apple and orange seeds). Peroxidases had attracted considerable interest in recent years because of their activities towards a wide variety of chromogenic substances. Peroxidase activity in crude extract of apple and orange seeds was measured by recording a spectrophotometric value. Partial purification of crude enzyme extract was done by ammonium sulfate precipitation and ion exchange chromatography. It was observed that after partial purification, the enzyme activity was increased as compared to crude enzyme extract. Peroxidase from orange seed was purified up to 17.17 fold with specific activity of 10.17 U/mg and that from apple seed was 6.82 fold with specific activity of 7.53 U/mg after diethyl amino ethyl ( DEAE) cellulose chromatography. It was shown that orange seed peroxidase had more activity than apple seed peroxidase in crude extract and each step of purification. Further purification was obtained through gel filtration chromatography by using sephadex-G-75 column. Peroxidase from orange and apple seeds got purified up to 30.64 and 8.34 fold with their specific activity of 18.16 and 9.20 U/mg, respectively. It is more evident that peroxidase is the most heat stable enzyme; therefore, it is concluded that it may be potentially useful for industrial purposes.