Purification and characterization of a heat stable ferredoxin isolated from Clostridium thermocellum.
A thermostable ferredoxin was purified from Clostridium thermocellum. The final preparation was homogeneous as judged by electrophoresis in sodium dodecyl sulfate polyacrylamide gel and sedimentation equilibrium. It contains eight atoms of iron and eight acid-labile sulfur groups per molecule, the molecular weight is estimated to be 6 400 and the isoelectric point 3.35. Its amino-acid composition is characterized by the absence of histidine residues and the presence of eight cysteine residues. The absorption spectrum has a maximum at 390 nm with a molar absorption coefficient of 39 x 10(3) M1 cm-1, similar to that of other bacterial eight iron ferredoxins. The purified ferredoxin has high thermal stability, since the spectrophotometric absorption of the protein at 390 nm did not change after one hour at 70 degrees C and only thirty five per cent of absorbance were lost after one hour at 80 degrees C. With regard to the electron carrier activity, the stability is slightly higher, only twenty five per cent of the activity were lost after one hour at 80 degrees C. During pyruvate oxidation, ferredoxin functions in the transfer of electrons to hydrogenase and also in the back reaction during pyridine nucleotide reduction by a ferredoxin -NAD oxidoreductase using hydrogen as electron donor.
D000596:Amino Acids / Q000032:analysis; D003013:Clostridium / Q000032:analysis*; D005288:Ferredoxins / Q000302:isolation & purification*; D006358:Hot Temperature; D007501:Iron / Q000032:analysis; D007526:Isoelectric Point; D008970:Molecular Weight; D009243:NAD / Q000378:metabolism; D013455:Sulfur / Q000032:analysis
D000596:Amino Acids; D005288:Ferredoxins; D009243:NAD; D013455:Sulfur; D007501:Iron