In vitro cell free synthesis of human manganese superoxide dismutase with the RTS 500 system

Reduced activity of manganese superoxide dismutase (MnSOD) is the basis of several pathologic features and complications occurring in the course of infectious mononucleosis. In order to supply future research with easily accessible enzyme, an in vitro protocol was developed based on the RTS 500 system and an overexpression vector. Translation of MnSOD monomers could be detected by SDS–PAGE, and assembly of the active homotetramer by native PAGE. Enzyme activity was successfully shown by in gel activity tests and enzyme assays. With 15 μg of DNA, 2.45 μkat were generated. The purification of MnSOD was performed by chromatography applying the His-tag technology. In SDS–PAGE of the eluate, a band showed up at Mr 25 000.