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Physiological heterogeneity of Pseudomonas taetrolens during lactobionic acid production

  • Biotechnological Products and Process Engineering
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Abstract

Physiological heterogeneity constitutes a critical parameter in biotechnological systems since both metabolite yield and productivity are often hampered by the presence of undesired physiological cell subpopulations. In the present study, the physiological status and functionality of Pseudomonas taetrolens cells were monitored by multiparameter flow cytometry during fermentative lactobionic acid production at the shake-flask and bioreactor scale. In shake-flask fermentation, the onset of the lactobionic acid production phase was accompanied by a progressive loss of cellular metabolic activity, membrane polarization, and membrane integrity concomitantly to acidification. In fact, population dynamics has shown the prevalence of damaged and dead subpopulations when submitted to a pH < 4 from 16 h onwards. Furthermore, fluorescence-activated cell sorting revealed that these sublethally injured cells were nonculturable. In contrast, P. taetrolens cells exhibited a robust physiological status during bioreactor cultivations performed with a pH-shifted strategy at 6.5, remaining predominantly healthy and metabolically active (>96 %) as well as maintaining bioconversion efficiency throughout the course of the fermentation. Additionally, an assessment of the seed culture’s physiological robustness was carried out in order to determine the best seed culture age. Results showed that bioreactor culture performance, growth, and lactobionic acid production efficiency were strongly dependent on the physiological heterogeneity displayed by the seed culture. This study provides the most suitable criteria for optimizing lactobionic acid production efficiency through a novel flow cytometric-based approach based on the physiological status of P. taetrolens. It also constitutes a valuable, broad-ranging methodology for the enhancement of microbial bioprocesses involved in the production of secondary metabolites.

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Acknowledgments

The authors are grateful for the financial support from the Spanish Ministry of Science and Innovation through project MEC-CTQ2010-14918. The technical assistance of Ana Salas, Magdalena Choda (Flow Cytometry Area, Scientific-Technical Services, University of Oviedo), and Marta Alonso (Process Image Area, Scientific-Technical Services, University of Oviedo) is gratefully acknowledged.

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Authors and Affiliations

  1. Department of Chemical Engineering and Environmental Technology, Faculty of Chemistry, University of Oviedo, C/ Julián Clavería s/n, 33071, Oviedo, Spain

    Saúl Alonso, Manuel Rendueles & Mario Díaz

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  1. Saúl Alonso
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  2. Manuel Rendueles
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  3. Mario Díaz
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Correspondence to Mario Díaz.

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Alonso, S., Rendueles, M. & Díaz, M. Physiological heterogeneity of Pseudomonas taetrolens during lactobionic acid production. Appl Microbiol Biotechnol 96, 1465–1477 (2012). https://doi.org/10.1007/s00253-012-4254-2

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  • DOI: https://doi.org/10.1007/s00253-012-4254-2

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