This study describes the feasibility and optimization of reactive dyeing on bio treated cotton knitted fabrics. For this, cotton knitted fabrics distinctly with two different enzymes, alkaline Pectinases(Scourzyme $L^{(R)}$) and Pectate lyases(Bactosol Co. ip $liquor^{(R)}$). In this way by increasing the concentration and processing temperature, the access of enzymes towards the fatty and waxy substrate was found to be accelerated. To achieve higher absorbency and whiteness index, a series of experiments was carried out to assure that Pectate lyases enzymes possesses high access towards the fats and waxes at high temperature. To this end, cotton knitted fabrics was dyed without oxidative bleaching step. The Pectate lyases scoured and dyed fabrics showed less color difference when 2% dye shade is used. The fabrics pre-scoured with Pectate lyases showed good the light and washing fastness properties, compared to the conventional and Pectinases dyed fabrics. However pectinases enzymes showed lower activity at high temperature, caused poor wettability and whiteness index of fabrics. The improvement of the accessibility of enzyme to the pectin at higher temperature Pectate lyases treatment before dyeing was found to be useful for subsequent pectin degradation in cotton knitted fabrics.