Enhanced Butanol Production by Free and Immobilized Clostridium sp. Cells Using Butyric Acid as Co-Substrate

Abstract: Butanol production by four different Clostridium sp. strains was investigated using glucoseP2-medium supplemented with increasing concentrations of butyric acid, added as cosubstrate.Batch fermentations were carried out in serum bottles (freely-suspended cellcultures) and fibrous-bed bioreactor (FBB) with medium recirculation (immobilized cells).Butyric acid clearly revealed to inhibit cellular growth with all specific growth rates decliningupon the increase of butyrate concentrations. However, the presence of low and moderatelevels in the medium can readily enhance the ABE-fermentation and increase butanolproduction through a shift induction towards the solventogenic phase controlled by themedium pH. In all cases it was found that 4.0 g⋅l-1 is the optimal concentration of butyratethat maximizes the yields for all ABE-solvents and butanol productivities. The non-mutant C.acetobutylicum ATCC 824 was singled out as the most efficient butanol productive strainamong all bacteria tested (10.3 g⋅l-1 butanol versus 0.72 g⋅l-1 with and without 4.0 g⋅l-1butyrate, respectively) showing a productivity augment in the order of 0.078 g⋅l-1⋅h-1 (78.5%)and yields of 0.3 g⋅g-1 from substrate and 7.6 g⋅g-1 from biomass versus 0.072 g⋅g-1 and 0.41g⋅g-1 with and without the optimal butyrate concentration, respectively. This strain alsorevealed the best overall tolerance over increasing butyrate concentrations up to ∼6.0 g⋅l-1 andthe highest glucose uptake (65.5%) among all bacteria. Furthermore, the beneficial effects ofbutyric acid were also observed through the use of a fibrous bed-bioreactor when the mutatedstrains of C. beijerinckii ATCC 55025 and BA 101 were tested. The use of this immobilizedcell system effectively improved butanol production over the free system with butanol titersin the fermentation broth around 11.5 g⋅l-1 and 9.4 g⋅l-1 for the two bacteria, respectively,roughly doubling the values attained with the corresponding suspended cell cultures when themedia were supplemented with 4.0 g⋅l-1 of butyrate. All these results confirm theenhancement of butanol formation using either free or immobilized cell culturessupplemented with butyric acid concentrations up to 4.0 g⋅l-1 in the media.