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Bacillus subtilis Endospores at High Purity and Recovery Yields: Optimization of Growth Conditions and Purification Method

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Abstract

Bacillus subtilis endospores have applications in different fields including their use as probiotics and antigen delivery vectors. Such specialized applications frequently require highly purified spore preparations. Nonetheless, quantitative data regarding both yields and purity of B. subtilis endospores after application of different growth conditions and purification methods are scarce or poorly reported. In the present study, we conducted several quantitative and qualitative analyses of growth conditions and purification procedures aiming generation of purified B. subtilis spores. Based on two growth media and different incubations conditions, sporulation frequencies up to 74.2 % and spore concentrations up to 7 × 109 spores/ml were achieved. Application of a simplified spore isolation method, in which samples were incubated with lysozyme and a detergent, resulted in preparations with highly purified spores at the highest yields. The present study represents, therefore, an important contribution for those working with B. subtilis endospores for different biotechnological purposes.

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Acknowledgments

The present study was supported by the Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Grants. We gratefully acknowledge Dr. A. L. Sonenshein, Tufts University, for the critical reading the manuscript and helpful suggestions. We also acknowledge the skillful technical assistance of L.C. Silva.

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Authors and Affiliations

  1. Vaccine Development Laboratory, Department of Microbiology, Institute of Biomedical Sciences, University of São Paulo, Avenida Lineu Prestes, 1374, São Paulo, SP, 05508-900, Brazil

    Milene B. Tavares, Renata D. Souza, Wilson B. Luiz, Caroline Casaroli, Eduardo G. Martins, Rita C. C. Ferreira & Luís C. S. Ferreira

  2. Department of Parasitology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil

    Rafael C. M. Cavalcante

Authors
  1. Milene B. Tavares
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  2. Renata D. Souza
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  3. Wilson B. Luiz
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  4. Rafael C. M. Cavalcante
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  5. Caroline Casaroli
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  6. Eduardo G. Martins
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  7. Rita C. C. Ferreira
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  8. Luís C. S. Ferreira
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Corresponding author

Correspondence to Luís C. S. Ferreira.

Additional information

Milene B. Tavares and Renata D. Souza contributed equally to this study.

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Tavares, M.B., Souza, R.D., Luiz, W.B. et al. Bacillus subtilis Endospores at High Purity and Recovery Yields: Optimization of Growth Conditions and Purification Method. Curr Microbiol 66, 279–285 (2013). https://doi.org/10.1007/s00284-012-0269-2

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  • DOI: https://doi.org/10.1007/s00284-012-0269-2

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