Two distinct bacterial strains were combined in a mixed culture to investigate the degradation of phenol. The mixed cultures consisting of species like Rhodococcus and Bacillus, were isolated from the waste effluent collected from Dankuni coal complex (DCC) in West Bengal. The experimental phenol biodegradation were performed batch wise at an optimized temperature and pH conditions of 37°C and 7.0 for a wide range of phenol concentrations initially, with saline solutions which is not supplemented with nutrients initially. Experimental results concluded a highest possible specific growth rate at 150 mg/L for phenol biodegradation of 2400 mg/L within a span of 72 h. Several significant physicochemical parameters were investigated in order to determine the ideal conditions for phenol breakdown. It was also observed that a spectroscopic study of the intermediate at 260 nm of the phenol biodegradation commenced through the formation of a cis, cis intermediate, indicating ortho‐cleavage pathway. A simulated substrate utilization and growth profiles was obtained by using substrate inhibition model equation using MATLAB R2017a were used to compare for the validation of the obtained experimental growth and substrate characterization data.