Long-Term Continuous Cultivation of Clostridium beijerinckii in a Two-Stage Chemostat with On-Line Solvent Removal

Abstract

A two-stage continuous cultivation experiment with Clostridium beijerinckii NRRL B592 is described. This strain maintained its ability to produce neutral solvents (acetone, n-butanol, and ethanol) at an overall dilution rate of 0.13 h(sup-1) and achieved an average overall solvent concentration of 9.27 g/liter and an overall solvent productivity of 1.24 g/liter/h for more than 100 overall retention times. The experiment was performed without pH control on a semisynthetic medium containing yeast extract, and product inhibition was the limiting factor. Solid carrier material was present in both stages, and the solvent productivity in both stages was similar. A membrane evaporation module integrated into the recirculation loop of a second-stage bioreactor after 2,166 h increased solvent productivity and improved the yield of solvents by about 40%. The membrane reduced the concentration of solvents, which would otherwise inhibit the fermentation. Additionally, the integrated membrane evaporation dampened metabolic oscillations, which are characteristic of continuous cultivation of clostridia. It was also demonstrated that a moderate concentration buildup (approximately 30% of bioreactor inflow) caused by water flux through the membrane caused no detrimental effects to the bacterial cells. However, much higher water fluxes through the membrane, associated with a much more dramatic increase in the concentration of salts in the medium, did appear to favor cell degeneration.