Autotrophic growth and CO2 fixation of Chloroflexus aurantiacus

Abstract

Chlorofluexus aurantiacus OK-70 fl was grown photoautotrophically with hydrogen as the electron source. The lowest doubling time observed was 26 h. The mechanism of CO2 fixation in autotrophically grown cells was studied. The presence of ribulose-1,5-bis-phosphate carboxylase and phosphoribulokinase could not be demonstrated. Carbon isotope fractionation (δ13C) was small, and alanine and aspartate but not 3-phosphoglycerate were the major labelled compounds in short term 14CO2 labelling. Thus CO2 is not fixed by the Calvin cycle. Fluoroacetate (FAc) completely inhibited protein synthesis in cultures and caused a slight citrate accumulation. However, CO2 fixation continued and increased polyglucose formation occurred. Under these conditions added acetate was metabolized to polyglucose, as were glycine, serine, glyoxylate and succinate, but to a lesser extent; little or no formate or CO was utilised. Glyoxylate inhibited CO2 fixation in vivo, indicating that pyruvate is formed from acetyl-CoA and CO2 by pyruvate synthase. Two key enzymes of the reductive TCA cycle, citrate lyase and α-ketoglutarate synthase were not detected in cell free extracts, but pyruvate synthase and phosphoenolpyruvate carboxylase were demonstrated. It is concluded that acetyl-CoA is a central intermediate in the CO2 fixation process, but the mechanism of its synthesis is not clear.