This study evaluated hydrogen production by co-culture of Ethanoligenens harbinense B49 and immobilized Rhodopseudomonas faecalis RLD-53 with different control strategies. To enhance cooperation of dark and photo-fermentation bactena during hydrogen production process, the glucose concentration, phosphate buffer concentration and initial pH were controlled at 6 g/l, 50 mmol/l and 7 5, respectively. The maximum yield and rate of hydrogen production were 3 10 mol H(2)/mol glucose and 17 2 mmol H(2)/l/h, respectively Ethanol from E harbinense B49 in acetate medium can enhance hydrogen production by R faecalis RLD-53 except the ratio of ethanol to acetate (R(E/A)) among 0 8 to 10 Control of the proper phosphate buffer concentration (SO mmol/l) not only increased acetic acid production by E harbinense B49, but also maintained stable pH of co-culture system Therefore, the results showed that co-culture of E harbinense B49 and immobilized R faecalis RLD-53 was a promising way of converting glucose into hydrogen (C) 2009 Professor T Nejat Veziroglu Published by Elsevier Ltd All rights reserved