This study was conducted to establish optimal temperature, pH and incubation time for bacteria from gin trash, for enhancing wax removal during cotton bioscouring. Universal Rotatable Experimental Design was used for the process of optimisation and efficiency assessed by the percentage weight loss after solvent extraction of the treated fibres. Optimum wax of 0.765% out of 1.2% present in the fibres was removed at 45°C, pH 8 within 1 h. Minitab 15 and Monte Carlos were used for the parameter modelling and sensitivity analysis. A second-order polynomial regression model was fitted and found adequate with a determination coefficient, R 2 , of 0.94032 (p < 0.005). At 45°C, pH 8, maximum 0.747% wax was removed by the bacteria in pectinase treatment within 1 h. 0.752% wax was removed from bulkier treated fibres, without significant change in fibre strength. Temperature was found to be the most significant variable in the bacterial cotton wax removal.