Pseudomonas putida KT2440 endures temporary oxygen limitations

Biotechnology and Bioengineering, vol. 118, pp. 4735–4750

Abstract

The obligate aerobic nature of Pseudomonas putida, one of the most prominent whole-cell biocatalysts emerging for industrial bioprocesses, questions its ability to be cultivated in large-scale bioreactors, which exhibit zones of low dissolved oxygen tension. P. putida KT2440 was repeatedly subjected to temporary oxygen limitations in scale-down approaches to assess the effect on growth and an exemplary production of rhamnolipids. At those conditions, the growth and production of P. putida KT2440 were decelerated compared to well-aerated reference cultivations, but remarkably, final biomass and rhamnolipid titers were similar. The robust growth behavior was confirmed across different cultivation systems, media compositions, and laboratories, even when P. putida KT2440 was repeatedly exposed to dual carbon and oxygen starvation. Quantification of the nucleotides ATP, ADP, and AMP revealed a decrease of intracellular ATP concentrations with increasing duration of oxygen starvation, which can, however, be restored when re-supplied with oxygen. Only small changes in the proteome were detected when cells encountered oscillations in dissolved oxygen tensions. Concluding, P. putida KT2440 appears to be able to cope with repeated oxygen limitations as they occur in large-scale bioreactors, affirming its outstanding suitability as a whole-cell biocatalyst for industrial-scale bioprocesses.

Authors 7

1. Philipp Demling Aachen Institute of Applied Microbiology

   RWTH Aachen University

   Affiliation as printed

   Institute of Applied Microbiology (iAMB), Aachen Biology and Biotechnology (ABBt) RWTH Aachen University Aachen Germany

   Institute of Applied Microbiology (iAMB), Aachen Biology and Biotechnology (ABBt), RWTH Aachen University, Aachen, Germany

2. Andreas Ankenbauer

   University of Stuttgart

   Affiliation as printed

   Institute of Biochemical Engineering University of Stuttgart Stuttgart Germany

   Institute of Biochemical Engineering, University of Stuttgart, Stuttgart, Germany

3. Bianca Klein

   Forschungszentrum Jülich

   Affiliation as printed

   Institute of Bio‐ and Geosciences (IBG‐1): Biotechnology Forschungszentrum Jülich GmbH Jülich Germany

4. Stephan Noack

   Forschungszentrum Jülich

   Affiliation as printed

   Institute of Bio‐ and Geosciences (IBG‐1): Biotechnology Forschungszentrum Jülich GmbH Jülich Germany

5. Till Tiso corresponding Aachen Institute of Applied Microbiology

   RWTH Aachen University

   Affiliation as printed

   Institute of Applied Microbiology (iAMB), Aachen Biology and Biotechnology (ABBt) RWTH Aachen University Aachen Germany

   Institute of Applied Microbiology (iAMB), Aachen Biology and Biotechnology (ABBt), RWTH Aachen University, Aachen, Germany

6. Ralf Takors

   University of Stuttgart

   Affiliation as printed

   Institute of Biochemical Engineering University of Stuttgart Stuttgart Germany

   Institute of Biochemical Engineering, University of Stuttgart, Stuttgart, Germany

7. Lars M. Blank corresponding Aachen Institute of Applied Microbiology

   RWTH Aachen University

   Affiliation as printed

   Institute of Applied Microbiology (iAMB), Aachen Biology and Biotechnology (ABBt) RWTH Aachen University Aachen Germany

   Institute of Applied Microbiology (iAMB), Aachen Biology and Biotechnology (ABBt), RWTH Aachen University, Aachen, Germany