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Characterization and Reconstitute of a [Fe4S4] Adenosine 5′-Phosphosulfate Reductase from Acidithiobacillus ferrooxidans

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Abstract

Adenosine 5′-phosphosulfate (APS) reductase is a key enzyme involved in the pathways of sulfate reduction and sulfide oxidation in the biological sulfur cycle. In this study, the gene of APS reductase from Acidithiobacillus ferrooxidans was cloned and expressed in Escherichia coli, the soluble protein was purified by one-step affinity chromatography to apparent homogeneity. The molecular mass of the recombinant APS reductase was determined to be 28 kDa using SDS-PAGE. According to optical and EPR spectra results of the recombinant protein confirmed that the iron–sulfur cluster inserted into the active site of the protein. Site-directed mutation for the enzyme revealed that Cys110, Cys111, Cys193, and Cys196 were in ligation with the iron–sulfur cluster. The [Fe4S4] cluster could be assembled in vitro, and exhibited electron transport and redox catalysis properties. As we know so far, this is the first report of expression in E. coli of APS reductase from A. ferrooxidans.

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Acknowledgments

This work was supported by the National Basic Research Program of People’s Republic of China (2004CB619204) and National Natural Science Foundation of People’s Republic of China (50321402) and FANEDD (200549).

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Authors and Affiliations

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

    Chunli Zheng, Yanfei Zhang, Yuandong Liu, Anna Wu, Lexian Xia, Jia Zeng, Jianshe Liu & Guanzhou Qiu

  2. Department of Biological and Chemical Engineering, Inner Mongolia University of Science and Technology, Baotou, 014010, People’s Republic of China

    Chunli Zheng

  3. College of Environmental Science and Engineering, Donghua University, Shanghai, 201620, People’s Republic of China

    Jianshe Liu

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  1. Chunli Zheng
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  2. Yanfei Zhang
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  3. Yuandong Liu
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  4. Anna Wu
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  5. Lexian Xia
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  6. Jia Zeng
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  7. Jianshe Liu
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  8. Guanzhou Qiu
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Correspondence to Jianshe Liu.

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Zheng, C., Zhang, Y., Liu, Y. et al. Characterization and Reconstitute of a [Fe4S4] Adenosine 5′-Phosphosulfate Reductase from Acidithiobacillus ferrooxidans . Curr Microbiol 58, 586–592 (2009). https://doi.org/10.1007/s00284-009-9375-1

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  • DOI: https://doi.org/10.1007/s00284-009-9375-1

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