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Analysis of two distinct mycelial populations in liquid-grown Streptomyces cultures using a flow cytometry-based proteomics approach

  • Genomics, transcriptomics, proteomics
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Abstract

Streptomycetes are proficient producers of enzymes and antibiotics. When grown in bioreactors, these filamentous microorganisms form mycelial pellets that consist of interconnected hyphae. We here employed a flow cytometry approach designed for large particles (COPAS) and demonstrate that liquid-grown Streptomyces cultures consist of two distinct populations of pellets. One population consists of mycelia with a constant mean diameter of approximately 260 μm, whereas the other population contains larger mycelia whose diameter depends on the strain, the age of the culture, and medium composition. Quantitative proteomics analysis revealed that 37 proteins differed in abundance between the two populations of pellets. Stress-related proteins and biosynthetic proteins for production of the calcium-dependent antibiotic were more abundant in the population of large mycelia, while proteins involved in DNA topology, modification, or degradation were overrepresented in the population of small mycelia. Deletion of genes for the cellulose synthase-like protein CslA and the chaplins affected the average size of the population of large pellets but not that of small pellets. Considering the fact that the production of enzymes and metabolites depends on pellet size, these results provide new leads toward rational strain design of Streptomyces strains tailored for industrial fermentations.

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Acknowledgments

MLCP and DC were appointed at IBL from the award to Prof. Dr. P.J.J. Hooykaas as Academy Professor. Bogdan Florea and Hermen Overkleeft are thanked for help with mass spectrometry. HABW and GPvW gratefully acknowledge the Dutch Applied Research Council (STW) for continuing financial support

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Authors and Affiliations

  1. Department of Microbiology, Institute of Biomembranes, University of Utrecht, Padualaan 8, 3584 CH, Utrecht, The Netherlands

    G. Jerre van Veluw & Han A. B. Wösten

  2. Molecular Biotechnology, Institute Biology Leiden (IBL), Leiden University, Sylviusweg 72, 2300 RA, Leiden, The Netherlands

    Marloes L. C. Petrus, Richard de Graaf, Inez P. de Jong, Gilles P. van Wezel & Dennis Claessen

  3. Department of Molecular and Developmental Genetics, Institute Biology Leiden (IBL), Leiden University, Sylviusweg 72, 2300 RA, Leiden, The Netherlands

    Marloes L. C. Petrus, Richard de Graaf, Inez P. de Jong & Dennis Claessen

  4. Molecular Biotechnology, Leiden Institute of Chemistry (LIC), Leiden University, Einsteinweg 55, 2333 CC, Leiden, The Netherlands

    Jacob Gubbens

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  1. G. Jerre van Veluw
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  2. Marloes L. C. Petrus
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  7. Han A. B. Wösten
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  8. Dennis Claessen
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Correspondence to Dennis Claessen.

Additional information

G. Jerre van Veluw, Marloes L.C. Petrus, and Jacob Gubbens contributed equally to the work.

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van Veluw, G.J., Petrus, M.L.C., Gubbens, J. et al. Analysis of two distinct mycelial populations in liquid-grown Streptomyces cultures using a flow cytometry-based proteomics approach. Appl Microbiol Biotechnol 96, 1301–1312 (2012). https://doi.org/10.1007/s00253-012-4490-5

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