Título
Expression and characterization of a recombinant psychrophilic Cu/Zn superoxide dismutase from deschampsia antarctica E. Desv. [Poaceae]
Autor
JUAN ANTONIO ROJAS CONTRERAS
Ana Paulina Barba de la Rosa
Antonio de León Rodríguez
Nivel de Acceso
Acceso Abierto
Identificador alterno
doi: https://doi.org/10.1007/s12010-015-1496-3
Materias
Resumen o descripción
"We present here the structural modeling and biochemical characterization of a recombinant superoxide dismutase (SOD) from Deschampsia antarctica E. Desv. [Poaceae] produced in Escherichia coli. The recombinant protein was purified by affinity chromatography nickel-nitrilotriacetic acid (Ni-NTA), and its identity was demonstrated by immunoblotting and inhibition by H2O2 and KCN. Inductively coupled plasma optical emission spectroscopy (ICP-OES) analysis confirmed the presence of Cu and Zn. Modeling of the D. antarctica Cu/Zn-SOD (DaSOD) amino acid sequence using the SWISS-MODEL and 2Q2L_B monomer of the psychrophilic Cu/Zu-SOD from Potentilla atrosanguinea (PaSOD) as template produced a structure similar to that of the typical eukaryotic Cu/Zn-SODs. Activity assays using the p-nitro blue tetrazolium chloride (NBT) solution method showed that the purified DaSOD had a specific activity of 5818 U/mg at 25 °C and pH 7.2 and that it was active in a pH interval of 5–8 and a temperature interval of 0–40 °C. Furthermore, DaSOD was still active at −20 °C as observed by a zymogram assay. We found 100 % activity when it was heated at 80 °C for 60 min, indicating a high thermostability. DaSOD properties suggest that this enzyme could be useful for preventing the oxidation of refrigerated or frozen foods, as well as in the preparation of cosmetic and pharmaceutical products."
Editor
Springer
Fecha de publicación
abril de 2015
Tipo de publicación
Artículo
Versión de la publicación
Versión enviada
Recurso de información
Formato
application/pdf
Relación
&
De León-Rodríguez, A. Appl Biochem Biotechnol (2015) 175: 3287. https://doi.org/10.1007/s12010-015-1496-3
Sugerencia de citación
Rojas-Contreras, J.A., de la Rosa, A.P.B.
Repositorio Orígen
Repositorio IPICYT
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