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Pakistan Journal of Biological Sciences
Year: 2000  |  Volume: 3  |  Issue: 3  |  Page No.: 407 - 410

Purification and Characterization of Polyphenoloxidase from Guava Infected with Fruit-rot Disease

M.A. Razzaque, Z.A. Saud, N. Absar, M.R. Karim and F. Hashinaga    

Abstract: Polyphenoloxidase (PPO), an important oxidative enzyme which played a vital role in the brewing of guava after infection with fruit-rot disease was purified from disease-affected guava. The method was accomplished by ion exchange chromatography of crude enzyme extract on DEAE-cellulose followed by CM-cellulose. The purified enzyme was found to be homogeneous as judged by polyacrylamide disc gel electrophoresis. Molecular weight of the enzyme was estimated to be 178 kDa and 180.5 kDa by gel filtration and SDS-PAGE respectively. The enzyme appeared to be a single polypeptide chain as revealed by SDS-PAGE either in presence or in absence of reducing agent. Optimum activity of the enzyme was at 48°C and pH around 8. Activity was destroyed completely in presence of EDTA, a metal chellator. Purified PPO showed an estimated km value of 4.4 mM for catechol as substrate.

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