The characterization of a thermostable and cambialistic superoxide dismutase from Thermus filiformis

Autor(es)

Mandelli, F. ^{[1, 2]} ; Franco Cairo, J. P. L. ^[1] ; Citadini, A. P. S. ^[1] ; Buechli, F. ^[1] ; Alvarez, T. M. ^[1] ; Oliveira, R. J. ^[3] ; Leite, V. B. P. ^[3] ; Paes Leme, A. F. ^[4] ; Mercadante, A. Z. ^[2] ; Squina, F. M. ^[1]

Afiliação do(s) autor(es)

^[1] CNPEM, Lab Nacl Ciencia & Tecnol Bioetanol CTBE, BR-13083970 Campinas, SP - Brazil

^[2] Univ Estadual Campinas, Fac Engn Alimentos, Dept Ciencia Alimentos, Campinas, SP - Brazil

^[3] Univ Estadual Paulista, Inst Biociencias Letras & Ciencias Exatas, Dept Fis, Sao Jose Do Rio Preto - Brazil

^[4] CNPEM, Lab Nacl Biociencias LNBio, BR-13083970 Campinas, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Letters in Applied Microbiology; v. 57, n. 1, p. 40-46, JUL 2013.
• Citações Web of Science: 10

Resumo

The 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 degrees C. The amount of enzyme required to inhibit 50% of pyrogallol autoxidation was 0 center dot 41, 0 center dot 56 and 13 center dot 73mg at 65, 70 and 80 degrees C, respectively. According to the circular dichroism (CD) spectra data, the secondary structure was progressively lost after increasing the temperature above 70 degrees C. The 3-dimensional model of TfSOD with the predicted cofactor binding corroborated with functional and CD analysis. Significance and Impact of the Study This manuscript describes the expression and characterization of a superoxide dismutase (SOD) from Thermus filiformis with thermophilic and cambialistic characteristics. The SODs are among the most potent antioxidants known in nature, and their stability and pharmacokinetics can vary widely in accordance to their biological source. Although the currently clinical research work has been focused on human and bovine SODs, alternative sources may become more biotechnological attractive in the near future. Our study brings new insights for the research field of antioxidant enzymes with potential application on pharmaceutical, cosmetics and food formulations. (AU)

• Processo FAPESP: 08/58037-9 - Geração de biblioteca para conversão enzimática de biomassa a partir de metagenoma do solo
• Beneficiário: Fábio Márcio Squina
• Modalidade de apoio: Auxílio à Pesquisa - Programa BIOEN - Jovens Pesquisadores
• Processo FAPESP: 11/17658-3 - Estudos computacionais em enovelamento de proteínas e aplicações no estudo de enzimas envolvidas na geração de bioetanol
• Beneficiário: Vitor Barbanti Pereira Leite
• Modalidade de apoio: Auxílio à Pesquisa - Regular