• Clostridium tyrobutyricum;
  • butyric acid;
  • two-phase partitioning bioreactor (TPPB);
  • in situ product recovery (ISPR);
  • carbon dioxide


Production of organic acids in solid–liquid two-phase partitioning bioreactors (TPPBs) is challenging, and highly pH-dependent, as cell growth occurs near neutral pH, while acid sorption occurs only at low pH conditions. CO2 sparging was used to achieve acidic pH swings, facilitating undissociated organic acid uptake without generating osmotic stress inherent in traditional acid/base pH control. A modified cultivation medium was formulated to permit greater pH reduction by CO2 sparging (pH 4.8) compared to typical media (pH 5.3), while still possessing adequate nutrients for extensive cell growth. In situ product recovery (ISPR) of butyric acid (pKa = 4.8) produced by Clostridium tyrobutyricum was achieved through intermittent CO2 sparging while recycling reactor contents through a column packed with absorptive polymer Hytrel® 3078. This polymer was selected on the basis of its composition as a polyether copolymer, and the use of solubility parameters for predicting solute polymer affinity, and was found to have a partition coefficient for butyric acid of 3. Total polymeric extraction of 3.2 g butyric acid with no CO2 mediated pH swings was increased to 4.5 g via CO2-facilitated pH shifting, despite the buffering capacity of butyric acid, which resists pH shifting. This work shows that CO2-mediated pH swings have an observable positive effect on organic acid extraction, with improvements well over 150% under optimal conditions in early stage fermentation compared to CO2-free controls, and this technique can be applied other organic acid fermentations to achieve or improve ISPR. Biotechnol. Bioeng. 2014;111: 537–544. © 2013 Wiley Periodicals, Inc.