In the present study, expression levels of nitrogenase encoding nifH and control genes nifA and prrA were examined at different physiological conditions in Rhodobacter sphaeroides O.U.001. In addition to variations in expression levels, changes in hydrogen production and growth were also investigated in response to different concentrations of ammonium source, acetate and aerobic conditions. In the present study, increasing concentration of ammonium chloride was found to be caused decrease in hydrogen production. Glutamate containing medium had the capacity for higher hydrogen production. Hydrogen production was observed even in aerobic conditions. The expression levels of nifH and nifA genes decreased with the increasing concentrations of ammonium chloride. Although the expression of nifA was present in the highest concentrations of NH(4)Cl under anaerobic conditions, no expression was observed under aerobic conditions of the same culture conditions. This was likely due to transcriptional level inhibition of nitrogenase in the presence of ammonium ion. Negative correlation was observed between the expression levels of prrA gene and its target, nifA gene. (C) 2009 Professor T. Nejat Veziroglu. Published by Elsevier Ltd. All rights reserved.