Fed-batch strategies using butyrate for high cell density cultivation of Pseudomonas putida and its use as a biocatalyst

Appl Microbiol Biotechnol. 2014 Nov;98(22):9217-28. doi: 10.1007/s00253-014-5989-8. Epub 2014 Aug 8.

Abstract

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(-1)over the lifetime of the fermentation, with a maximum mcl-PHA accumulation of 65 % of the cell dry weight.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Batch Cell Culture Techniques
  • Biomass
  • Biotransformation
  • Butyrates / metabolism*
  • Carbon / metabolism
  • Decanoates / metabolism
  • Enzymes*
  • Models, Theoretical
  • Polyhydroxyalkanoates / metabolism
  • Pseudomonas putida / enzymology*
  • Pseudomonas putida / growth & development*

Substances

  • Butyrates
  • Decanoates
  • Enzymes
  • Polyhydroxyalkanoates
  • Carbon