Studies on autotrophic CO2 fixation by the filamentous anoxygenic photosynthetic bacterium Oscillochloris trichoides strain DG-6 demonstrated that, unlike other green bacteria, this organism metabolized CO2 via the reductive pentose phosphate cycle. Both key enzymes of this cycle--ribulose-1,5-bisphosphate carboxylase/oxygenase and phosphoribulokinase--were detected in cell extracts. The main product of ribulose 1,5-bisphosphate-dependent CO2 fixation was 3-phosphoglyceric acid. KCN, which is known to be a competitive inhibitor of ribulose-1,5-bisphosphate carboxylase/oxygenase, completely inhibited the CO2 assimilation by whole cells as well as by cell extracts of O. trichoides. The 13C/12C carbon isotope fractionation during photoautotrophic growth of O. trichoides was -19.7/1000, which is close to that obtained for autotrophic organisms that use ribulose-1,5-bisphosphate carboxylase as the primary carboxylation enzyme. Cell extracts of O. trichoides contained all the enzymes of the tricarboxylic acid cycle except 2-oxoglutarate dehydrogenase. No activity of isocitrate lyase, a key enzyme of the glyoxylate shunt, was found in cell extracts of O. trichoides DG-6.