**2.4 Precipitation using ammonium sulphate**

The crude enzymes of the isolates was treated with ammonium sulphate (analytical grade, Sigma) within 40–90% saturation. Precipitation was at 4°C for 24 hours. Centrifugation was done at 4000 rpm for 30 minutes at 4°C using a high speed cold centrifuge. The precipitate was re-constituted in 0.2 M citrate phosphate buffer (pH 5.0). Protein content of the precipitated enzyme was determined [22]. Polygalacturonase activity was also determined [23, 24].
