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The plant extract was done in an ice-cold buffer containing 50 mM phosphate buffer (pH 6.5) containing 0.1% triton X, 0.2 mM EDTA, 1% (W/V) polyvinylpyrrolidone (PVP), 1% leupeptine, 0.1% DTT, 1 mM proteinase K, 5 mM ascorbate and centrifuged at 10,000 g at 4 C for 20 min