The bacterial isolate, Pseudomonas aeruginosa BR was tested for its electrochemical activity in different zones within the anode chamber of the microbial fuel cell (MFC). Zonal activity was studied using cyclic voltammetry (CV), impedance spectroscopy (EIS) and fluorescent spectroscopy. Redox activity was prominent and charge transfer impedance (CT) was 13.94 Omega and 58.64 Omega referring to the flow of electron from microbe to biofilm via mediator in the anode chamber (Z1 zone). The abode biofilm (Z2 zone) depicted less redox activity and CT of 48.79 Omega. The anolyte (Z3 zone) had the lowest CT of 20.13 Omega and moderate redox activity. The bacterial cells were 69.18 % and 91.64 % live in anode biofilm and anolyte respectively. The culture was prominently active in the anolyte zone of the MFC. This awareness of zonal activity can be applicable in the integrated MFC systems where targeted action of microbes can be an added benefit.