Microorganisms actively catabolize substrate, and bioelectricity is generated. Microbial fuel cells (MFCs) could be utilized as a power generator in small devices. The discovery of species of microorganisms is called Rhodopseudomonas palustris KU-EGAT 13. The experiments have been conducted with the production of electricity from this type of microorganism in a single chamber microbial fuel cell. The study used 4 surface roughness of anode electrode single chamber micro fuel to study the performance of microbial fuel cell effect from the anode. Three stainless steel plates were polished to uniform roughness to the magnitude between 0.05 and 1 µm. After 24 h of experimentation, the rough electrode’s open-circuit voltage (OCV) and power densities were much higher than that produced by the smooth one. Moreover, the smooth surface is higher than the charge-transfer resistance of the rough electrode. The rough surface’s better electrochemical performance is due to denser biofilm grown on the surface, which was observed by scanning electron microscopy (SEM) and figuring out the microbial number in an image using an ImageJ program. HIGHLIGHTS The new discovery of species of microorganisms is called Rhodopseudomonas palustris KU-EGAT 13 which can produce electricity in a single chamber microbial fuel cell The performance of MFCs has gotten an effect from the different anode surface roughness.The biomass growth is larger than a rougher anode surface, which contributes to anode’s efficiency After observed by SEM, found that when it is in the highest anode surface roughness directed towards the number of microbes stick on the surface GRAPHICAL ABSTRACT