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Molecular characterization and expression analysis of iron superoxide dismutase gene from Pseudochlorella pringsheimii (Trebouxiophyceae, Chlorophyta)

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Abstract

The FeSOD isoforms of Pseudochlorella pringsheimii were identified, a preliminary characterization of the enzyme was conducted, and the relationship among the FeSOD gene from P. pringsheimii and that of other organisms was examined. The FeSOD has an open reading frame of 612 bp that encodes 203 deduced amino acids with a molecular mass of 23 kDa. Expression of the recombinant FeSOD gene was done successfully in Escherichia coli. The purified FeSOD has a specific enzyme activity that reached 688 U mg−1 protein (in vitro assay). Alkaline conditions showed the highest activity for the recombinant FeSOD. Moreover, it showed a relative thermostability up to 50 °C, while at 50 and 70 °C, the activity was reduced by 32 and 68%, respectively, after 1 h as compared to the maximum. Phylogenetic analysis revealed three main clusters i.e., the prokaryotic Cyanophyta, bacteria, and the eukaryotic Chlorophyta intermingled with plant species and a dinoflagellate. P. pringsheimii was closely grouped with Chlorella pyrenoidosa, however, other species showed a relative disparity. Alignment of FeSOD gene sequences of the different species showed many conserved regions which could be used for FeSOD sequences among unexplored species and may be useful for the taxonomy of the revised coccoid Chlorella species.

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Acknowledgements

The authors thank Drs. G. Hausner and M.H. Abdelfattah (University of Manitoba, MB, Canada), for technical assistance. The first author would like to thank the Ministry of Higher Education and Scientific Research (MHESR, Egypt) for the financial support (through a post doctoral fellowship). This work was financially assisted by a Natural Sciences and Engineering Research Council (NSERC) grant (to Dr. Piercey-Normore).

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Authors and Affiliations

  1. Department of Biological Sciences, Faculty of Science, University of Manitoba, Winnipeg, MB, R3T 2N2, Canada

    Mostafa M. S. Ismaiel & Michele D. Piercey-Normore

  2. Botany Department, Faculty of Science, Zagazig University, Zagazig, 44519, Egypt

    Mostafa M. S. Ismaiel

  3. School of Science and the Environment, Memorial University of NL (Grenfell Campus), Corner Brook, NL, A2H 5G4, Canada

    Michele D. Piercey-Normore

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  1. Mostafa M. S. Ismaiel
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Correspondence to Mostafa M. S. Ismaiel.

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Ismaiel, M.M.S., Piercey-Normore, M.D. Molecular characterization and expression analysis of iron superoxide dismutase gene from Pseudochlorella pringsheimii (Trebouxiophyceae, Chlorophyta). Physiol Mol Biol Plants 25, 221–228 (2019). https://doi.org/10.1007/s12298-018-0569-5

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