Recovery Yields of Protopectinase Depending on Treatments of Organic Solvents

유기용매의 처리에 따른 Bacillus subtillis IFO 12113 유래 Protopectinase의 회수

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.

Protopectinase(PPase)는 식물 세포벽의 protopectin을 분해하는 효소로서 식품의 가공에 이용될 수 있는 효소이다. PPase의 산업적 응용을 위하여 PPase를 생산하는 미생물 Bacillus subtilis IFO 12113의 배양여액에 acetone, methanol, ethanol을 각각 처리하여 PPase를 회수하는 연구를 수행하였다. Acetone의 경우, 배양여액 대 acetone을 1:1 (v/v) 비율로 첨가했을 때 효소는 59.2%가 회수되었으며 정제도는 1.7배 이었다. Methanol의 경우, 1:2(v/v) 비율로 처리하였을 때, 효소는 100% 회수되었으며 비활성도로 본 정제도는 4.0배 증가하였다. Ethanol의 경우에 1 :0.5 (v/v) 비율로 처리한 것을 원심분리시켜 침전물을 제거하고, 그 상등액에 최종 ethanol 비율을 1:1(v/v)까지 처리했을 때, 효소의 회수율은 68%를 나타냈으며 비활성도는 13.5배 증가하였다. 세균의 배양여액으로부터 PPase의 회수에는 methanol 처리가 가장 효율적이었으나, 산업적 응용을 위하여 정제효과까지 고려하면 ethanol처리가 가장 적절하였다.