Abstract—This study investigates the use of two carbon sources in fermentation for the production of cellobiohydrolase (CBH), an adaptive or inducible enzyme. Selection of carbon source was first done using single substrate cultivation between glucose, maltose and lactose using recombinant A. niger PY11. This recombinant microorganism will produce CBH in high quantity, with maltose acting as its main inducer. Glucose and maltose were selected in dual carbon fermentation, as glucose produce highest biomass 0.32g/ml at day 4^th and maltose produce 0.42g/ml biomass at day 9^th. Cultivation of A. niger was done for dual carbon containing sugar glucose and maltose with concentration ratio 1:1. Cultivation with medium containing maltose was done for comparison between single carbon and dual carbon. For dual carbon cultivation, highest biomass form at day 7^th at 0.44g/ml while specific enzyme activity reached 10.54 U/g at day 8^th. For single substrate, highest biomass production was recorded at day 9^th with 0.41g/ml.while specific enzyme activity at day 7^th at 4.02 U/g.