Palm Oil Mill Effluent (POME) is an effluent that has been discharge in a high organic content condition after palm oil production. POME has been recognized as a controversial wastewater because of the strong abilities to initiate environmental damages. Characterization of pollutants characteristic in POME was identified and biogas was produced from mixed culture of POME in batch treatment. It could be an escape plan to reduce the potential impact to environment. Three sample of ponding treatment system was obtained in Felda Kemahang Palm Oil Factory and the samples is POME from raw pond, facultative pond and anaerobic pond. Characteristic of POME can be analyze in Total Suspended Solid (TSS), Total Solid (TS), Biological Oxygen Demand (BOD) and water quality that include Total Dissolved Solid (TDS), Dissolved Oxygen (DO), Salinity and Conductivity value. Different glucose concentration (0, 20, 40) was added in the synthetic medium for fermentation process. Different glucose concentration will show which concentration can rapidly process the biogas reaction. Mixed culture inoculation shows the difference growth development which is high growth in anaerobic POME pond (control concentration) and raw POME pond (20, 40 glucose concentration). Biogas was produced in each glucose concentration and 20 g glucose concentration show the highest production especially in facultative POME pond. The factors affecting the biogas production is by effects of temperature and concentration of glucose.