In the present study, semi-IPN membranes of sulfonated styrene (SS) and sulfonated PVdF-co-HFP membranes have been analyzed as polymer electrolyte membrane in single chamber microbial fuel cells (MFCs). A respective 5%, 10%, 20% and 30% sulfonated styrene (SS) with varying concentrations of divinyl benzene (DVB) have been polymerized in the presence of sulfonated PVdF-co-HFP to 10 prepare SPS-0(0% SS), SPS-5 (5% SS), SPS-10(10% SS), SPS-20(20% SS) and SPS-30(30% SS) membranes. Progressive improvements in membrane properties was observed with increasing SS and DVB concentration, where excess DVB (< 0.8 wt % of SS) resulted in increased crosslinks within membrane structure. This eventually impeded membrane property and altered rigidity in membranes. The membranes were characterized for their ion exchange capacity (IEC) and proton conductivity; IEC value of 0.39 meq g -1 , 0.42 meq g -1 , 0.47 meq g -1 , 0.54 meq g -1 and 0.63 meq g -1 and proton conductivity of 3.23×10 -3 , 1.06×10 -2 , 1.87×10 -2 , 2.47×10 -2 and 15 1.61×10 -2 Scm -1 were observed for SPS-0(0% SS), SPS-5 (5% SS), SPS-10(10% SS), SPS-20(20% SS) and SPS-30(30% SS) membranes. The membranes were sandwiched as membrane electrode assemblies (MEA) and employed in single chambered MFCs with open air cathode to analyze their overall performance output. It was observed that amongst these membranes, MFC with SPS-20 membrane showed the maximum power and current density of 447.42 ± 22 mW/m 2 and 1729.63 ± 87mA/m 2 with an overall ~91.27% COD removal in 28 days operation, using electrogenic mixed firmicutes as biocatalysts. In overall, the study depicts the relevance of 20 semi-IPN membranes of sulfonated styrene (SS) and sulfonated PVdF-co-HFP in MFC application for harvesting bio-energy.