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A highly thermostable ferritin from the hyperthermophilic archaeal anaerobe Pyrococcus furiosus

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Abstract

A ferritin from the obligate anaerobe and hyperthermophilic archaeon Pyrococcus furiosus (optimal growth at 100°C) has been cloned and overproduced in Escherichia coli to one-fourth of total cell-free extract protein, and has been purified in one step to homogeneity. The ferritin (PfFtn) is structurally similar to known bacterial and eukaryal ferritins; it is a 24-mer of 20 kDa subunits, which add up to a total Mr 480 kDa. The protein belongs to the non-heme type of ferritins. The 24-mer contains approximately 17 Fe (as isolated), 2,700 Fe (fully loaded), or <1 Fe (apoprotein). Fe-loaded protein exhibits an EPR spectrum characteristic for superparamagnetic core formation. At 25°C Vmax=25 μmole core Fe3+ formed per min per mg protein when measured at 315 nm, and the K0.5=5 mM Fe(II). At 0.3 mM Fe(II) activity increases 100-fold from 25 to 85°C. The wild-type ferritin is detected in P. furiosus grown on starch. PfFtn is extremely thermostable; its activity has a half-life of 48 h at 100°C and 85 min at 120°C. No apparent melting temperature was found up to 120°C. The extreme thermostability of PfFtn has potential value for biotechnological applications.

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Acknowledgements

We are grateful to prof. Simon de Vries for helpful discussion. We thank prof. Ana Maria Varela Coelho from the ITQB (Oeiras, Portugal) for providing mass spectrometry data. Anton Korteweg from Wageningen University (The Netherlands) is acknowledged for help with differential scanning calorimetry. This research has been financially supported by the Council for Chemical Sciences of the Netherlands Organization for Scientific Research (CW-NWO) under project number 700.51.301.

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  1. Department of Biotechnology, Delft University of Technology, Julianalaan 67, 2628 BC, Delft, The Netherlands

    Jana Tatur, Peter-Leon Hagedoorn, Marieke L. Overeijnder & Wilfred R. Hagen

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  1. Jana Tatur
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  2. Peter-Leon Hagedoorn
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  3. Marieke L. Overeijnder
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  4. Wilfred R. Hagen
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Correspondence to Wilfred R. Hagen.

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Communicated by F. Robb

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Tatur, J., Hagedoorn, PL., Overeijnder, M.L. et al. A highly thermostable ferritin from the hyperthermophilic archaeal anaerobe Pyrococcus furiosus. Extremophiles 10, 139–148 (2006). https://doi.org/10.1007/s00792-005-0484-x

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