Abstract: The objective of this study was to investigate hydrogen production from alcohol wastewater by anaerobic sequencing batch reactor (ASBR) under thermophillic operation. The ASBR unit used in this study had a liquid holding volume of 4 L and was operated at 6 cycles per day. The seed sludge taken from an upflow anaerobic sludge blanket unit treating the same wastewater was boiled at 95 °C for 15 min before being fed to the ASBR unit. The ASBR system was operated at different COD loading rates at a thermophillic temperature (55 °C), and controlled pH of 5.5. When the system was operated under optimum conditions (providing maximum hydrogen production performance) at a feed COD of 60 000 mg/l, and a COD loading rate of 68 kg/m3 d, the produced gas contained 43 % H2 content in the produced gas. Moreover, the hydrogen yield and the specific hydrogen production rate (SHPR) were 130 ml H2/g COD removed and 2100 ml H2/l d, respectively.