The effects of the various carbon and nitrogen substrates on the growth and polygalacturonase activity of Trichoderma viride (BITRS-1001) isolated from the tar sand deposit in Gbelejuloda-Irele Ondo State, Nigeria were investigated in submerged cultivation at 30 °C ± 2 °C. The commercial carbon and nitrogen substrates included sucrose, fructose, starch, maltose, lactose and peptone, sodium nitrate, urea and casein respectively. All the carbon substrates used supported the growth of T. viride (0.566 to 0.156 g/50 mL of culture medium) with starch supporting the highest biomass yield and sucrose the least biomass yield. Maximum polygalacturonase activity of 3033 U/mL was recorded in maltose medium. Maximum biomass yield on the nitrogen sources was observed in the organic nitrogen namely peptone and casein with values not significantly different from each other at p ≤ 0.05. In the determination of the crude enzyme activity on the nitrogen sources, maximum polygalacturonase activity of 12,400 U/mL was recorded in peptone medium. Hence, a careful manipulation of these nutrient substrates could help to optimise the production of this enzyme on a large scale.