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Characterization of recombinant glutathione reductase from the psychrophilic Antarctic bacterium Colwellia psychrerythraea

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Abstract

Glutathione reductases catalyze the reduction of oxidized glutathione (glutathione disulfide, GSSG) using NADPH as the substrate to produce reduced glutathione (GSH), which is an important antioxidant molecule that helps maintain the proper reducing environment of the cell. A recombinant form of glutathione reductase from Colwellia psychrerythraea, a marine psychrophilic bacterium, has been biochemically characterized to determine its molecular and enzymatic properties. C. psychrerythraea glutathione reductase was shown to be a homodimer with a molecular weight of 48.7 kDa using SDS-PAGE, MALDI-TOF mass spectrometry and gel filtration. The C. psychrerythraea glutathione reductase sequence shows significant homology to that of Escherichia coli glutathione reductase (66 % identity), and it possesses the FAD and NADPH binding motifs, as well as absorption spectrum features which are characteristic of flavoenzymes such as glutathione reductase. The psychrophilic C. psychrerythraea glutathione reductase exhibits higher k cat and k cat/K m at lower temperatures (4 °C) compared to mesophilic Baker’s yeast glutathione reductase. However, C. psychrerythraea glutathione reductase was able to complement an E. coli glutathione reductase deletion strain in oxidative stress growth assays, demonstrating the functionality of C. psychrerythraea glutathione reductase over a broad temperature range, which suggests its potential utility as an antioxidant enzyme in heterologous systems.

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Abbreviations

C. psychrerythraea :

Colwellia psychrerythraea

GR:

Glutathione reductase

GSH:

Reduced glutathione

GSSG:

Oxidized glutathione

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Acknowledgments

Support for this study was provided by NASA’s Institute for Advanced Concepts (NIAC) and by the North Carolina Agriculture Research Station. The authors would like to thank Jason Whitham and Dr. Sherry Tove for assistance in editing the manuscript and Janet Kim for help with statistical analysis.

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Authors and Affiliations

  1. Department of Plant and Microbial Biology, North Carolina State University, 4550A Thomas Hall Campus Box 7612, Raleigh, NC, 27695, USA

    Mikyoung Ji & Amy M. Grunden

  2. Department of Animal Science, North Carolina State University, 123 Polk Hall Campus Box 7621, Raleigh, NC, 27695, USA

    Callie V. Barnwell

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  1. Mikyoung Ji
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  2. Callie V. Barnwell
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Correspondence to Amy M. Grunden.

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Communicated by H. Atomi.

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Ji, M., Barnwell, C.V. & Grunden, A.M. Characterization of recombinant glutathione reductase from the psychrophilic Antarctic bacterium Colwellia psychrerythraea . Extremophiles 19, 863–874 (2015). https://doi.org/10.1007/s00792-015-0762-1

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