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Expression and Characterization of a Recombinant Psychrophilic Cu/Zn Superoxide Dismutase from Deschampsia antarctica E. Desv. [Poaceae]

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Abstract

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.

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Acknowledgments

This work was partially financed by CONACyT-Básicas Grant No. 178988. Juan Rojas thanks CONACyT for his scholarship No. 204213. The authors thank Leandro G. Ordoñez for technical support and Jennifer Ecklerly for English correction. We thank the Chilean Antarctic Institute (INACH) for the logistic support during the stay in the Scientific Base “Prof. Julio Escudero,” King George Island, Antarctic.

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Authors and Affiliations

  1. División de Biología Molecular, Instituto Potosino de Investigación Científica y Tecnológica, A.C. (IPICyT), Camino a la Presa San José 2055, Lomas 4a. Sección, CP 78216, San Luis Potosí, Mexico

    Juan A. Rojas-Contreras, Ana P. Barba de la Rosa & Antonio De León-Rodríguez

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  1. Juan A. Rojas-Contreras
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  2. Ana P. Barba de la Rosa
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  3. Antonio De León-Rodríguez
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Correspondence to Antonio De León-Rodríguez.

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Rojas-Contreras, J.A., de la Rosa, A.P.B. & De León-Rodríguez, A. Expression and Characterization of a Recombinant Psychrophilic Cu/Zn Superoxide Dismutase from Deschampsia antarctica E. Desv. [Poaceae]. Appl Biochem Biotechnol 175, 3287–3296 (2015). https://doi.org/10.1007/s12010-015-1496-3

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