The research was conducted with the production of pectinase enzyme by fungal fermentation using Rhizopus oryzae and characterization of produced enzyme with respect to pH, temperature, incubation time, and substrate specificity. Carbon source was optimized replacing sugar with different concentration of pectin (0.5, 0.75, and 1.0%) during submerged fermentation. The outcomes of the fermentation process showed that media containing carbon source of 1% pectin replaced dextrose at pH 6.0 incubating for 72 hours at 35 °C were the best condition for pectinase production. The maximal activity for pectinase enzyme produced from Rhizopus oryzae by fermentation was 3.16 U.mL-1 and it was found at 40 °C and pH 6.5. The produced pectinase enzyme was found thermo stable up to 60 °C for 50 min. The activity of the enzyme was increased with the increasing pectin concentration in the media and maximum activity was found at the pectin concentration of 5 mg.mL-1. The km and Vmax values were found 0.84 mg.mL-1 and 5.294 mg.mL-1 respectively at optimized condition. The outcomes of the research may be useful for further research in low cost production of pectinases from R.oryzae for domestic consumption in many industries.