4.6 Article

High Cell Density Cultivation of Pseudomonas putida KT2440 Using Glucose Without the Need for Oxygen Enriched Air Supply

Journal

BIOTECHNOLOGY AND BIOENGINEERING
Volume 112, Issue 4, Pages 725-733

Publisher

WILEY
DOI: 10.1002/bit.25474

Keywords

bioprocess engineering; modeling; Pseudomonas putida KT2440; fed-batch; glucose; polyhydroxyalkanoates

Funding

  1. Enterprise Ireland
  2. Industrial Development Agency

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High Cell Density (HCD) cultivation of bacteria is essential for the majority of industrial processes to achieve high volumetric productivity (gL(-1) h(-1)) of a bioproduct of interest. This study developed a fed batch bioprocess using glucose as sole carbon and energy source for the HCD of the well described biocatalyst Pseudomonas putida KT2440 without the supply of oxygen enriched air. Growth kinetics data from batch fermentations were used for building a bioprocess model and designing feeding strategies. An exponential followed by linearly increasing feeding strategy of glucose was found to be effective in maintaining biomass productivity while also delaying the onset of dissolved oxygen (supplied via compressed air) limitation. A final cell dry weight (CDW) of 102gL(-1) was achieved in 33h with a biomass productivity of 3.1gL(-1) h(-1) which are the highest ever reported values for P. putida strains using glucose without the supply of pure oxygen or oxygen enriched air. The usefulness of the biomass as a biocatalyst was demonstrated through the production of the biodegradable polymer polyhydroxyalkanoate (PHA). When nonanoic acid (NA) was supplied to the glucose grown cells of P. putida KT2440, it accumulated 32% of CDW as PHA in 11h (2.85gL(-1) h(-1)) resulting in a total of 0.56kg of PHA in 18L with a yield of 0.56g PHA g NA(-1). Biotechnol. Bioeng. 2015;112: 725-733. (c) 2014 Wiley Periodicals, Inc.

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