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Expression, Purification and Molecular Structure Modeling of Thioredoxin (Trx) and Thioredoxin Reductase (TrxR) from Acidithiobacillus ferrooxidans

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Abstract

The thioredoxin system consists of thioredoxin (Trx), thioredoxin reductase (TrxR) and NADPH, which plays several key roles in maintaining the redox environment of the cell. In Acidithiobacillus ferrooxidans, thioredoxin system may play important functions in the activity regulation of periplasmic proteins and energy metabolism. Here, we cloned thioredoxin (trx) and thioredoxin reductase (trxR) genes from Acidithiobacillus ferrooxidans, and expressed the genes in Escherichia coli. His-Trx and His-TrxR were purified to homogeneity with one-step Ni-NTA affinity column chromatography. Site-directed mutagenesis results confirmed that Cys33, Cys36 of thioredoxin, and Cys142, Cys145 of thioredoxin reductase were active-site residues.

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Acknowledgements

This study was supported by the National Basic Research Program of P. R. China (2004CB619204), and the National Natural Science Foundation of P. R. China (50621063).

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Authors and Affiliations

  1. Department of Bioengineering, School of Resources Processing and Bioengineering, Central South University, Changsha, Hunan, 410083, People’s Republic of China

    Yiping Wang, Xiaojian Zhang, Qing Liu, Chenbing Ai, Hongyu Mo & Jia Zeng

  2. Biomedical Engineering Center, Hunan University, Changsha, Hunan, 410082, People’s Republic of China

    Jia Zeng

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  1. Yiping Wang
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  2. Xiaojian Zhang
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  3. Qing Liu
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  4. Chenbing Ai
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  5. Hongyu Mo
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  6. Jia Zeng
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Correspondence to Jia Zeng.

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Wang, Y., Zhang, X., Liu, Q. et al. Expression, Purification and Molecular Structure Modeling of Thioredoxin (Trx) and Thioredoxin Reductase (TrxR) from Acidithiobacillus ferrooxidans . Curr Microbiol 59, 35–41 (2009). https://doi.org/10.1007/s00284-009-9390-2

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  • DOI: https://doi.org/10.1007/s00284-009-9390-2

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