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Cloning, Expression, Purification, and Characterization of Glutaredoxin from Antarctic Sea-Ice Bacterium Pseudoalteromonas sp. AN178

Glutaredoxins (Grxs) are small ubiquitous redox enzymes that catalyze glutathione-dependent reactions to reduce protein disulfide. In this study, a full-length Grx gene (PsGrx) with 270 nucleotides was isolated from Antarctic sea-ice bacterium Pseudoalteromonas sp. AN178. It encoded deduced 89 amino...

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Autores principales: Wang, Quanfu, Hou, Yanhua, Shi, Yonglei, Han, Xiao, Chen, Qian, Hu, Zhiguo, Liu, Yuanping, Li, YuJin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Hindawi Publishing Corporation 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4109671/
https://www.ncbi.nlm.nih.gov/pubmed/25110664
http://dx.doi.org/10.1155/2014/246871
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author Wang, Quanfu
Hou, Yanhua
Shi, Yonglei
Han, Xiao
Chen, Qian
Hu, Zhiguo
Liu, Yuanping
Li, YuJin
author_facet Wang, Quanfu
Hou, Yanhua
Shi, Yonglei
Han, Xiao
Chen, Qian
Hu, Zhiguo
Liu, Yuanping
Li, YuJin
author_sort Wang, Quanfu
collection PubMed
description Glutaredoxins (Grxs) are small ubiquitous redox enzymes that catalyze glutathione-dependent reactions to reduce protein disulfide. In this study, a full-length Grx gene (PsGrx) with 270 nucleotides was isolated from Antarctic sea-ice bacterium Pseudoalteromonas sp. AN178. It encoded deduced 89 amino acid residues with the molecular weight 9.8 kDa. Sequence analysis of the amino acid sequence revealed the catalytic motif CPYC. Recombinant PsGrx (rPsGrx) stably expressed in E. coli BL21 was purified to apparent homogeneity by Ni-affinity chromatography. rPsGrx exhibited optimal activity at 30°C and pH 8.0 and showed 25.5% of the activity at 0°C. It retained 65.0% of activity after incubation at 40°C for 20 min and still exhibited 37.0% activity in 1.0 M NaCl. These results indicated that rPsGrx was a typical cold active protein with low thermostability.
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spelling pubmed-41096712014-08-10 Cloning, Expression, Purification, and Characterization of Glutaredoxin from Antarctic Sea-Ice Bacterium Pseudoalteromonas sp. AN178 Wang, Quanfu Hou, Yanhua Shi, Yonglei Han, Xiao Chen, Qian Hu, Zhiguo Liu, Yuanping Li, YuJin Biomed Res Int Research Article Glutaredoxins (Grxs) are small ubiquitous redox enzymes that catalyze glutathione-dependent reactions to reduce protein disulfide. In this study, a full-length Grx gene (PsGrx) with 270 nucleotides was isolated from Antarctic sea-ice bacterium Pseudoalteromonas sp. AN178. It encoded deduced 89 amino acid residues with the molecular weight 9.8 kDa. Sequence analysis of the amino acid sequence revealed the catalytic motif CPYC. Recombinant PsGrx (rPsGrx) stably expressed in E. coli BL21 was purified to apparent homogeneity by Ni-affinity chromatography. rPsGrx exhibited optimal activity at 30°C and pH 8.0 and showed 25.5% of the activity at 0°C. It retained 65.0% of activity after incubation at 40°C for 20 min and still exhibited 37.0% activity in 1.0 M NaCl. These results indicated that rPsGrx was a typical cold active protein with low thermostability. Hindawi Publishing Corporation 2014 2014-07-07 /pmc/articles/PMC4109671/ /pubmed/25110664 http://dx.doi.org/10.1155/2014/246871 Text en Copyright © 2014 Quanfu Wang et al. https://creativecommons.org/licenses/by/3.0/ This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Wang, Quanfu
Hou, Yanhua
Shi, Yonglei
Han, Xiao
Chen, Qian
Hu, Zhiguo
Liu, Yuanping
Li, YuJin
Cloning, Expression, Purification, and Characterization of Glutaredoxin from Antarctic Sea-Ice Bacterium Pseudoalteromonas sp. AN178
title Cloning, Expression, Purification, and Characterization of Glutaredoxin from Antarctic Sea-Ice Bacterium Pseudoalteromonas sp. AN178
title_full Cloning, Expression, Purification, and Characterization of Glutaredoxin from Antarctic Sea-Ice Bacterium Pseudoalteromonas sp. AN178
title_fullStr Cloning, Expression, Purification, and Characterization of Glutaredoxin from Antarctic Sea-Ice Bacterium Pseudoalteromonas sp. AN178
title_full_unstemmed Cloning, Expression, Purification, and Characterization of Glutaredoxin from Antarctic Sea-Ice Bacterium Pseudoalteromonas sp. AN178
title_short Cloning, Expression, Purification, and Characterization of Glutaredoxin from Antarctic Sea-Ice Bacterium Pseudoalteromonas sp. AN178
title_sort cloning, expression, purification, and characterization of glutaredoxin from antarctic sea-ice bacterium pseudoalteromonas sp. an178
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4109671/
https://www.ncbi.nlm.nih.gov/pubmed/25110664
http://dx.doi.org/10.1155/2014/246871
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