Expression and characterization of a recombinant psychrophilic Cu/Zn superoxide dismutase from deschampsia antarctica E. Desv. [Poaceae]
"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...
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ftipicytsanjose:oai:repositorio.ipicyt.edu.mx:11627/3894 2023-05-15T13:54:02+02:00 Expression and characterization of a recombinant psychrophilic Cu/Zn superoxide dismutase from deschampsia antarctica E. Desv. [Poaceae] Rojas Contreras, Juan Antonio Barba de la Rosa, Ana Paulina De León Rodríguez, Antonio Springer 2015-04 application/pdf http://hdl.handle.net/11627/3894 https://doi.org/10.1007/s12010-015-1496-3 unknown Rojas-Contreras, J.A., de la Rosa, A.P.B. & De León-Rodríguez, A. Appl Biochem Biotechnol (2015) 175: 3287. https://doi.org/10.1007/s12010-015-1496-3 http://hdl.handle.net/11627/3894 https://doi.org/10.1007/s12010-015-1496-3 Attribution-NonCommercial-NoDerivatives 4.0 Internacional http://creativecommons.org/licenses/by-nc-nd/4.0/ Acceso Abierto CC-BY-NC-ND Antioxidant Extremophile Oxidative stress Psychrophilic enzyme Superoxide ion BIOLOGÍA MOLECULAR article 2015 ftipicytsanjose https://doi.org/10.1007/s12010-015-1496-3 2020-05-03T16:26:04Z "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." Article in Journal/Newspaper Antarc* Antarctica Repository IPICYT (Instituto Potosino de Investigación Científica y Tecnológica) Applied Biochemistry and Biotechnology 175 7 3287 3296 |
institution |
Open Polar |
collection |
Repository IPICYT (Instituto Potosino de Investigación Científica y Tecnológica) |
op_collection_id |
ftipicytsanjose |
language |
unknown |
topic |
Antioxidant Extremophile Oxidative stress Psychrophilic enzyme Superoxide ion BIOLOGÍA MOLECULAR |
spellingShingle |
Antioxidant Extremophile Oxidative stress Psychrophilic enzyme Superoxide ion BIOLOGÍA MOLECULAR Rojas Contreras, Juan Antonio Barba de la Rosa, Ana Paulina De León Rodríguez, Antonio Expression and characterization of a recombinant psychrophilic Cu/Zn superoxide dismutase from deschampsia antarctica E. Desv. [Poaceae] |
topic_facet |
Antioxidant Extremophile Oxidative stress Psychrophilic enzyme Superoxide ion BIOLOGÍA MOLECULAR |
description |
"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." |
author2 |
Springer |
format |
Article in Journal/Newspaper |
author |
Rojas Contreras, Juan Antonio Barba de la Rosa, Ana Paulina De León Rodríguez, Antonio |
author_facet |
Rojas Contreras, Juan Antonio Barba de la Rosa, Ana Paulina De León Rodríguez, Antonio |
author_sort |
Rojas Contreras, Juan Antonio |
title |
Expression and characterization of a recombinant psychrophilic Cu/Zn superoxide dismutase from deschampsia antarctica E. Desv. [Poaceae] |
title_short |
Expression and characterization of a recombinant psychrophilic Cu/Zn superoxide dismutase from deschampsia antarctica E. Desv. [Poaceae] |
title_full |
Expression and characterization of a recombinant psychrophilic Cu/Zn superoxide dismutase from deschampsia antarctica E. Desv. [Poaceae] |
title_fullStr |
Expression and characterization of a recombinant psychrophilic Cu/Zn superoxide dismutase from deschampsia antarctica E. Desv. [Poaceae] |
title_full_unstemmed |
Expression and characterization of a recombinant psychrophilic Cu/Zn superoxide dismutase from deschampsia antarctica E. Desv. [Poaceae] |
title_sort |
expression and characterization of a recombinant psychrophilic cu/zn superoxide dismutase from deschampsia antarctica e. desv. [poaceae] |
publishDate |
2015 |
url |
http://hdl.handle.net/11627/3894 https://doi.org/10.1007/s12010-015-1496-3 |
genre |
Antarc* Antarctica |
genre_facet |
Antarc* Antarctica |
op_relation |
Rojas-Contreras, J.A., de la Rosa, A.P.B. & De León-Rodríguez, A. Appl Biochem Biotechnol (2015) 175: 3287. https://doi.org/10.1007/s12010-015-1496-3 http://hdl.handle.net/11627/3894 https://doi.org/10.1007/s12010-015-1496-3 |
op_rights |
Attribution-NonCommercial-NoDerivatives 4.0 Internacional http://creativecommons.org/licenses/by-nc-nd/4.0/ Acceso Abierto |
op_rightsnorm |
CC-BY-NC-ND |
op_doi |
https://doi.org/10.1007/s12010-015-1496-3 |
container_title |
Applied Biochemistry and Biotechnology |
container_volume |
175 |
container_issue |
7 |
container_start_page |
3287 |
op_container_end_page |
3296 |
_version_ |
1766259552282476544 |