An efficient method for the expression and reconstitution of thermostable Mn/Fe superoxide dismutase from Aeropyrum pernix K1.
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.
Lee
Hee-Jin
HJ
Department of Chemistry, College of Natural Sciences, Chung-Ang University, Seoul, Korea.
Kwon
Hye-Won
HW
Koh
Jong-Uk
JU
Lee
Dong-Kuk
DK
Moon
Ja-Young
JY
Kong
Kwang-Hoon
KH
D041522:Aeropyrum / Q000201:enzymology* / Q000235:genetics; D000595:Amino Acid Sequence; D001483:Base Sequence; D019641:DNA, Archaeal / Q000737:chemistry / Q000235:genetics; D004789:Enzyme Activation; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D007218:Industrial Microbiology / Q000379:methods*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D016415:Sequence Alignment; D013482:Superoxide Dismutase / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification
D019641:DNA, Archaeal; D011994:Recombinant Proteins; D013482:Superoxide Dismutase