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Study of metabolism and identification of productive regions in filamentous fungi via spatially resolved time-of-flight secondary ion mass spectrometry

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Abstract

Filamentous fungi are well-established production hosts that feature a strong interconnection between morphology, physiology, and productivity. For penicillin production in Penicillium chrysogenum, industrial processes frequently favor a pellet morphology comprising compact hyphal agglomerates. Inherently these tightly packed entanglements lead to inactive, degrading sections within the pellet’s core because of limitations. Optimal process design requires detailed knowledge of the nature of the limitations and localization of productive zones in the biomass, which is generally obtainable through modeling and complex analytical methods such as oxygen microelectrode and histological investigations. Methods that combine physiological and morphological insight are crucial yet scarce for filamentous fungi. In this study, we used time-of-flight secondary ion mass spectrometry in combination with oxygen and glucose tracer substrates, requiring little effort for sample preparation and measurement. Our method is capable of analyzing oxygen and substrate uptake in various morphological structures by the use of 18O as a tracer. In parallel, we can assess productive biomass regions through identification of penicillin mass fragments to simultaneously study oxygen diffusion, substrate incorporation, and productive biomass sections.

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Acknowledgements

We thank the Austrian Federal Ministry of Science, Research and Economy through the Christian Doppler Laboratory for financial support. Strains for the experiments were gratefully provided by Sandoz (Kundl, Austria).

Funding

This study was funded by Austrian Federal Ministry of Science, Research and Economy through the Christian Doppler Laboratory for Mechanistic and Physiological Methods for Improved Bioprocesses (grant number 171).

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Authors and Affiliations

  1. Research Area Biochemical Engineering, Institute of Chemical, Environmental and Bioscience Engineering, Technische Universität Wien, Gumpendorfer Straße 1a, 1060, Vienna, Austria

    Lukas Veiter, Christoph Herwig & Christoph Slouka

  2. Christian Doppler Laboratory for Mechanistic and Physiological Methods for Improved Bioprocesses, Technische Universität Wien, Gumpendorfer Straße 1a, 1060, Vienna, Austria

    Lukas Veiter, Christoph Herwig & Christoph Slouka

  3. Institute of Chemical Technologies and Analytics, Technische Universität Wien, Getreidemarkt 9/164, 1060, Vienna, Austria

    Markus Kubicek, Herbert Hutter & Ernst Pittenauer

Authors
  1. Lukas Veiter
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  2. Markus Kubicek
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  3. Herbert Hutter
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  4. Ernst Pittenauer
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  5. Christoph Herwig
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  6. Christoph Slouka
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Contributions

LV and CS designed the study, performed the experiments, analyzed the data, and wrote the article. MK performed and analyzed the ToF-SIMS measurements. EP performed mass spectrometry measurements. HH provided valuable scientific input. CH supervised the work.

Corresponding author

Correspondence to Christoph Slouka.

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The authors declare that they have no competing interests.

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This article does not contain any studies with human participants or animals performed by any of the authors.

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Published in the topical collection Advances in Process Analytics and Control Technology with guest editor Christoph Herwig.

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Veiter, L., Kubicek, M., Hutter, H. et al. Study of metabolism and identification of productive regions in filamentous fungi via spatially resolved time-of-flight secondary ion mass spectrometry. Anal Bioanal Chem 412, 2081–2088 (2020). https://doi.org/10.1007/s00216-019-01980-2

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  • DOI: https://doi.org/10.1007/s00216-019-01980-2

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