In this study, the production of biogas from vinasse by using three SGBR reactors with the same volume of 5.6 liters for three hydraulic retention times (2, 3 and 4 days) and organic loading rates of 86002, 28667 and 21500 mgCOD/L.d was investigated. The water bath method was used to provide the temperature of mesophyll reactors at 35, and NaOH solution was used for buffer. The biogas volume was measured using the water displacement method. The results showed that the methane production was almost constant with HRT of 4, 3, and 2 days after 11, 13, and 16 days, respectively. Methane gas production with HRT of 2, 3, and 4 days was 6.66, 16.66 and 27.38 LCH4 (STP)/L.day per liter of vinasse, respectively. For three reactors, methanogen and acidogenic bacteria were in the equilibrium state at pH 5.57 after 2 days. The amount of methane gas production per volatile solids with the HRT of 2, 3, and 4 days was 244, 345 and 434 m3CH4/gvs, respectively.