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High cell density cultivation of the chemolithoautotrophic bacterium Nitrosomonas europaea

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Abstract

Nitrosomonas europaea is a chemolithoautotrophic nitrifier, a gram-negative bacterium that can obtain all energy required for growth from the oxidation of ammonia to nitrite, and this may be beneficial for various biotechnological and environmental applications. However, compared to other bacteria, growth of ammonia oxidizing bacteria is very slow. A prerequisite to produce high cell density N. europaea cultures is to minimize the concentrations of inhibitory metabolic by-products. During growth on ammonia nitrite accumulates, as a consequence, N. europaea cannot grow to high cell concentrations under conventional batch conditions. Here, we show that single-vessel dialysis membrane bioreactors can be used to obtain substantially increased N. europaea biomasses and substantially reduced nitrite levels in media initially containing high amounts of the substrate. Dialysis membrane bioreactor fermentations were run in batch as well as in continuous mode. Growth was monitored with cell concentration determinations, by assessing dry cell mass and by monitoring ammonium consumption as well as nitrite formation. In addition, metabolic activity was probed with in vivo acridine orange staining. Under continuous substrate feed, the maximal cell concentration (2.79 × 1012/L) and maximal dry cell mass (0.895 g/L) achieved more than doubled the highest values reported for N. europaea cultivations to date.

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Acknowledgments

The research was supported by the EU and cofinanced by the European Social Fund under the project ENVIKUT (TÁMOP-4.2.2.A-11/1/KONV-2012-0043) and by the Hungarian Scientific Research Fund (OTKA K1006600 and NN116519). The authors are grateful to Zoltán Németh, Zoltán Fekete, Antal Kökényesi, Csilla Noémi Lipták, and István Kolláth for their help. We thank József Kozma (Gedeon Richter Plc, Budapest, Hungary) for critically reading the manuscript.

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Authors and Affiliations

  1. Department of Biochemical Engineering, Faculty of Science and Technology, University of Debrecen, H-4032, Egyetem tér 1, Debrecen, Hungary

    Benedek Papp, Erzsébet Fekete, Michel Flipphi & Levente Karaffa

  2. Safety and Environmental Department, TEVA Pharmaceutical Ltd., H-4032, Pallagi u. 13, Debrecen, Hungary

    Tibor Török

  3. Institute of Food Science, Faculty of Agricultural and Food Sciences and Environmental Management, University of Debrecen, H-4032, Böszörményi út 138, Debrecen, Hungary

    Erzsébet Sándor

Authors
  1. Benedek Papp
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  2. Tibor Török
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  3. Erzsébet Sándor
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  4. Erzsébet Fekete
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  5. Michel Flipphi
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  6. Levente Karaffa
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Corresponding author

Correspondence to Levente Karaffa.

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Fig. S1

Microscopic image of struvite crystals formed in the early stages of N. europaea fermentations. For technical details, see Materials and methods section. (PDF 43 kb)

Fig. S2

Microscopic images of N. europaea cells stained with Acridine Orange at 20 (image 1.), 40 (2.), 70 (3.) and 80 h (4.) of the conventional batch fermentation presented in Fig. 3. For technical details, see Materials and methods section. (PDF 85 kb)

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Papp, B., Török, T., Sándor, E. et al. High cell density cultivation of the chemolithoautotrophic bacterium Nitrosomonas europaea . Folia Microbiol 61, 191–198 (2016). https://doi.org/10.1007/s12223-015-0425-8

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  • DOI: https://doi.org/10.1007/s12223-015-0425-8

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