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Development of a defined medium supporting rapid growth for Deinococcus radiodurans and analysis of metabolic capacities

  • Applied Microbial and Cell Physiology
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Abstract

A morpholinepropanesulfonic acid (MOPS)-buffered rich defined medium (RDM) was optimized to support a reproducible 2.6-h doubling time at 35 °C for Deinococcus radiodurans R1 and used to gain insight into vitamin and carbon metabolism. D. radiodurans was shown to require biotin and niacin for growth in this medium. A glutamine–serine simple defined medium (SDM) was developed that supported a 4-h doubling time, and this medium was used to probe sulfur and methionine metabolism. Vitamin B12 was shown to alleviate methionine auxotrophy, and under these conditions, sulfate was used as the sole sulfur source. Phenotypic characterization of a methionine synthase deletion mutant demonstrated that the B12 alleviation of methionine auxotrophy was due to the necessity of the B12-dependent methionine synthase in methionine biosynthesis. Growth on ammonium as the sole nitrogen source in the presence of vitamin B12 was demonstrated, but it was not possible to achieve reproducibly good growth in the absence of at least one amino acid as a nitrogen source. Growth on sulfate, cysteine, and methionine as sulfur sources demonstrated the function of a complete sulfur recycling pathway in this strain. These studies have demonstrated that rapid growth of D. radiodurans R1 can be achieved in a MOPS-based medium solely containing a carbon source, salts, four vitamins, and two amino acids.

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Acknowledgements

We thank Doug Pitera, Gregory Morin, and Natalia Korotkova for helpful discussions, Marina Kalyuzhnaya and Ludmila Chistoserdova for helpful comments on the manuscript. This work was supported by grants from the DOE EMSP program (ER20294), and the NSF Graduate Research Fellowship Program.

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Authors and Affiliations

  1. Department of Chemical Engineering, University of Washington, Seattle, WA, 98195-2180, USA

    Alexandra D. Holland, Heather M. Rothfuss & Mary E. Lidstrom

  2. Department of Microbiology, University of Washington, Seattle, WA, 98195-2180, USA

    Mary E. Lidstrom

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  1. Alexandra D. Holland
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  2. Heather M. Rothfuss
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  3. Mary E. Lidstrom
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Correspondence to Mary E. Lidstrom.

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Holland, A.D., Rothfuss, H.M. & Lidstrom, M.E. Development of a defined medium supporting rapid growth for Deinococcus radiodurans and analysis of metabolic capacities. Appl Microbiol Biotechnol 72, 1074–1082 (2006). https://doi.org/10.1007/s00253-006-0399-1

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  • DOI: https://doi.org/10.1007/s00253-006-0399-1

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