This work reports on production of ethanol with simultaneous fixing of nitrogen (N2) using the anaerobic bacterium Zymomonas mobilis DSM 473. A batch fermentation with an initial glucose concentration of 50 g L-1, an initial pH of ∼5.5, an inoculum size of 10% by volume and a N2 feeding rate of 50 mL min-1 without mechanical agitation was found to provide the highest ethanol productivity (0.401 g L-1 h-1). Ethanol yield on glucose exceeded 97% of the theoretical maximum. The nitrogen content of the microbial biomass was 10.4% w/w at 65 h and all of it was derived by fixation of dinitrogen. Repeated-batch fermentations were investigated for ethanol production using simultaneous nitrogen fixation. A 2-cycle repeated-batch fermentation lasting 71 h gave a maximum ethanol yield on glucose of 0.475 g g-1 and an ethanol productivity of 0.675 g L-1 h-1. The yield (0.415 g g-1) and productivity (0.638 g L-1 h-1) were reduced in a 3-cycle repeated batch operation lasting 94 h. The need to fix nitrogen did not reduce the final achievable ethanol concentration, or the ethanol yield on glucose, relative to fermentations provided with fixed nitrogen, but did reduce the ethanol productivity by ∼82% because less cell mass was produced.
Keywords: Ethanol; Nitrogen fixation; Repeated-batch fermentation; Zymomonas mobilis.
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