The kinetics and the effects of various parameters on hydrogen production by Rhodobacter sphaeroides O.U. 001 were investigated in a batch column photobioreactor. In particular, the effect of the inoculum age and the implementation of a light-dark cycle illumination scheme for emulating natural sunlight have been investigated in detail. The possibility of using yeast extract to replace the rather expensive vitamin mixture in the medium was also studied. The results show that hydrogen production is decreased when the initially inoculated bacteria have a high culture age. Exposure of the bacterial culture to light-dark cycles increased the amount of hydrogen compared to continuous illumination, all other parameters remaining the same. Similarly, the use of yeast extract to replace the vitamins increased the growth and hydrogen production rates, however, with a slight reduction in the total amount of gas produced and the hydrogen fraction in the evolved gas. (C) 2002 International Association for Hydrogen Energy. Published by Elsevier Science Ltd. All rights reserved.