In this work, microbial fuel cell (MFC) design of five chambers was used to investigate the effect of four types of membranes which are cation exchange membrane (CEM), Cellulose Triacetate membrane (CTA), thin film composite membrane (TFC), and proton exchange membrane (PEM). To study the influence of the membrane type on the cathode performance, the four cathode chambers were filled with 20 g/l NaCl catholyte and the sodium acetate of 1.5 g/l was supplied to the central chamber as anolyte. The results revealed that the membrane proton selectivity plays an important role in the cathodic reduction reaction for electrical generation and water production. It was observed that the PEM has a significant effect on the power generation with a maximum power density of 20.492 mW/m2 with water production of 4.21 g/day. Whereas the competition of the other cations to the proton transfer was clearly observed by using the CTA membrane with power production of 12.646 mW/m2, and the abundance of the water production of 178.16 g/day was attributed to the water transport across the CTA membrane. For studying the influence of the membrane type on the anode performance, the sodium acetate of 1.5 g/l was supplied to the four chambers as an anolyte at a flow rate of 0.0272 cm3/sec and the central chamber were filled with 20 g/l NaCl catholyte. The salt reverse transfer from the cathode chamber to the anode chamber across the CTA membrane contributed to increasing the anolyte electrical conductivity and consequently increased the power production to 12.555 mW/m2. Meanwhile, the effect of the proton selectivity and the electrical resistance of the other membrane were observed in the other chambers. Thus, the usage of CEM, TFC, and PEM produced electrical power of 6.751, 3.004, and 9.712 mW/m2 respectively.