A pectinesterase(PE) as a hydrolase was purified and partially characterized from pectinand cassava starch-supplemented wheat bran culture of Rhizopus japonicus IF05318, a strain selected among 25 isolates and authentic strains(from IFO). The PE was purified 162 folds with a recovery of 10.6% based on the total enzyme activity by batch extraction and column chromatography using SP-Sephadex C-50, CM-Sephadex C-50 and Sephadex G-75 gel filtration. The enzyme was stable at pH range of 5.0-8.5 and up to 40℃. The pH and temperature optima were 5.0 and 35℃, respectively. The K,,, value was estimated as small value as 18.4mgIml from the Lineweaver-Burk plot, comparing reported fungal K,,, values. The molecular weight was determined as 33kDa by SDS-PAGE. The activity enhancers were Cu^<2+>, Ca^<2+>, Na^+ and Mg^<2+> while the inhibitors were Pb^<2+>, Ba^<2+>, Zn^<2+>, Fe^<2+> and tannic acid.