Bio-hydrogen production and the F0F1-ATPase activity of Rhodobacter sphaeroides: Effects of various heavy metal ions

Rhodobacter sphaeroides MDC 6521 isolated from Arzni mineral springs in Armenia is able to produce bio-hydrogen (H₂) in anaerobic conditions upon illumination in the presence of various metal ions. The significant aspect in regulation of H₂ production by these bacteria and its energetics is the requirement for F₀F₁-ATPase, the main membrane enzyme responsible for generation of proton motive force under anaerobic conditions. In order to determine the mediatory role of F₀F₁ in H₂ production, the effects of various metal ions (Mn²⁺, Mg²⁺, Fe²⁺, Ni²⁺, and Mo⁶⁺) on N,N′-dicyclohexylcarbodiimide inhibited ATPase activity of R. sphaeroides membrane vesicles were investigated. These ions in appropriate concentrations considerably enhanced H₂ production, which was not observed in the absence of Fe²⁺, indicate the requirement for Fe²⁺. The R. sphaeroides membrane vesicles demonstrated significant ATPase activity. In the absence of Fe²⁺ inhibition (∼80%) of ATPase activity was observed, which was increased by addition of metal ions. A higher ATPase activity was detected in the presence of Fe²⁺ (80 μM) and Mo⁶⁺ (16 μM). These results indicate a relationship between the F₀F₁-ATPase activity and H₂ production that might be a significant pathway to provide novel evidence of a requirement for F₀F₁-ATPase in H₂ production by R. sphaeroides.