An efficient method for the expression and reconstitution of thermostable Mn/Fe superoxide dismutase from Aeropyrum
Hee-Jin Lee1, Hye-Won Kwon, Jong-Uk Koh
1Department of Chemistry, College of Natural Sciences, Chung-Ang University, Seoul, Korea.
Abstract:
The gene APE0743 encoding the superoxide dismutase (ApSOD) of a hyperthermophilic archaeon Aeropyrum pernix K1 was cloned and over-expressed as a GST fusion protein at a high level in Escherichia coli. The expressed protein was simply purified by the process of glutathione affinity chromatography and thrombin treatment. The ApSOD was a homodimer of 25 kDa subunits and a cambialistic SOD which was active with either Fe(II) or Mn(II) as a cofactor. The ApSOD was highly stable against high temperature. This thermostable ApSOD is expected to be applicable as a useful biocatalyst for medicine and bio-industrial processes.
