Extracellular Pectinase Production and Purification from a Newly Isolated Bacillus subtilis Strain

Abstract

Pectinase is widely used in the fruit juice industry with various aspects, especially to reduce blur in fruit juice. To produce pectinase, Bacillus subtilis were isolated from Kilis soil contaminated by agricultural crop wastes. The extracellular pectinase was purified by using three different technics (cell-free supernatant, ethanol, and ammonium sulphate precipitation) and enzyme activity was measured with spectrophotometric analysis. The maximum specific activity was observed in enzyme preparate purified by ethanol precipitation, 217.44 U/mg. The degradation products of reaction–glucose, saccharose, and fructose–were identified by Fourier transform infrared spectroscopy and thin layer chromatography analysis. Two protein bands belong to pectinase of B. subtilis were determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Molecular weights of bands were approximately calculated as 60 and 64 kDa.