Fed-batch strategies using butyrate for high cell density cultivation of Pseudomonas putida and its use as a biocatalyst
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A mathematically based fed-batch bioprocess demonstrated the suitability of using a relatively cheap and renewable substrate (butyric acid) for Pseudomonas putida CA-3 high cell density cultivation. Butyric acid fine-tuned addition is critical to extend the fermentation run and avoid oxygen consumption while maximising the biomass volumetric productivity. A conservative submaximal growth rate (μ of 0.25 h−1) achieved 71.3 g L−1 of biomass after 42 h of fed-batch growth. When a more ambitious feed rate was supplied in order to match a μ of 0.35 h−1, the volumetric productivity was increased to 2.0 g L−1 h−1, corresponding to a run of 25 h and 50 g L−1 of biomass. Both results represent the highest biomass and the best biomass volumetric productivity with butyrate as a sole carbon source. However, medium chain length polyhydroxyalkanoate (mcl-PHA) accumulation with butyrate grown cells is low (4 %). To achieve a higher mcl-PHA volumetric productivity, decanoate was supplied to butyrate grown cells. This strategy resulted in a PHA volumetric productivity of 4.57 g L−1 h−1 in the PHA production phase and 1.63 g L−1 h−1over the lifetime of the fermentation, with a maximum mcl-PHA accumulation of 65 % of the cell dry weight.
KeywordsHigh cell density Fed-batch bioprocess Butyric acid Biocatalyst Polyhydroxyalkanoate
Federico Cerrone was funded by Enterprise Ireland and the Irish Industrial Development Agency (IDA) through the Technology Center for Biorefining and Bioenergy (project no. CC20090004). The authors have no conflict of interest to declare.
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