Mandelli, F.Franco Cairo, J. P LCitadini, A. P SBüchli, F.Alvarez, T. M.Oliveira, R. J. [UNESP]Leite, V. B P [UNESP]Paes Leme, A. F.Mercadante, A. Z.Squina, F. M.2014-05-272014-05-272013-07-01Letters in Applied Microbiology, v. 57, n. 1, p. 40-46, 2013.0266-82541472-765Xhttp://hdl.handle.net/11449/75818The superoxide dismutase (TfSOD) gene from the extremely thermophilic bacterium Thermus filiformis was cloned and expressed at high levels in mesophilic host. The purified enzyme displayed approximately 25 kDa band in the SDS-PAGE, which was further confirmed as TfSOD by mass spectrometry. The TfSOD was characterized as a cambialistic enzyme once it had enzymatic activity with either manganese or iron as cofactor. TfSOD showed thermostability at 65, 70 and 80°C. The amount of enzyme required to inhibit 50% of pyrogallol autoxidation was 0·41, 0·56 and 13·73 mg at 65, 70 and 80°C, respectively. According to the circular dichroism (CD) spectra data, the secondary structure was progressively lost after increasing the temperature above 70°C. The 3-dimensional model of TfSOD with the predicted cofactor binding corroborated with functional and CD analysis. © 2013 The Society for Applied Microbiology.40-46eng3-D modelCambialistic enzymeCircular dichroismNitroblue tetrazoliumPyrogallol autoxidationironmanganesepyrogallolsuperoxide dismutasecloneenzyme activitygene expressioninhibitionmass spectrometrythermophilic bacteriumthree-dimensional modelingamino acid sequenceautooxidationcircular dichroismcontrolled studymolecular cloningnonhumanpolyacrylamide gel electrophoresisprotein expressionprotein purificationprotein secondary structurethermostabilityThermusThermus filiformisArtigo10.1111/lam.12071WOS:000320316600007Acesso restrito2-s2.0-848790292200500034174785796