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To recover protopectinase (PPase) secreted from Bacillus subtilis IFO 12113, culture filtrate of the microorganism was treated with acetone, methanol, and ethanol, respectively. In the case of treatment with acetone at a ratio of 1 : 1 (culture filtrate : acetone, v/v), PPase was purified 1.7-fold with 59.2% recovery. The recovery of PPase was increased by increasing the acetone concentration. PPase was purified 4-fold with 100% recovery when the culture filtrate was precipitated with methanol at a ratio of 1 : 2 (culture filtrate : methanol, v/v). However, recovery of PPase was decreased by increasing the methanol concentration. PPase was purified 13.5-fold resulting in 68% recovery by the addition of ethanol with the final ratio 1 : 1(culture filtrate : ethanol, v/v) to the supernatant, which was obtained after precipitation of the culture filtrate with ethanol at a ratio of 1 : 0.5. These results show that methanol treatment is better than other organic solvent treatments for the simple recovery of PPase, whereas fractionated treatment of ethanol can recover PPase with higher purification fold.
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