Abstract

Degenerate strains of Clostridium acetobutylicum lack the ability to produce solvents and sporulate and remain in a permanent acidogenic state, allowing continuous hydrogen and organic acid production through anaerobic fermentation. Eichhornia crassipes, an invasive aquatic plant, emerges as a promising source of fermentable sugars for hydrogen production via anaerobic fermentation. In this study, a degenerated strain of Clostridium acetobutylicum was isolated and subsequently cultivated in the presence of a hydrolysate solution obtained from the alkaline pre-treatment and enzymatic hydrolysis of Eichhornia crassipes. The hydrolysate was mixed with a defined medium and served the dual purpose of providing essential nutrients and mitigating inhibitors, eliminating the need for an additional detoxification step. A pure defined culture medium served as a control. The extraction methods employed led to the release of low concentrations of inhibitors, reaching 0.1 g/L of furfural and 0.18 g/L of HMF. Kinetic characterization revealed that in the presence of Eichhornia crassipes hydrolysate, the degenerate strain exhibited lower specific growth rates ranging from 0.114 to 0.156 h?1, compared with the control medium which ranged from 0.131 to 0.179 h?1. This was accompanied by lower yields, ranging from 0.115 to 0.167 gDCW/g in the presence of hydrolysate versus 0.178 to 0.190 gDCW/g in the control medium, and diminished butyric acid production of 1.318 to 2.932 g/L in the presence of hydrolysate versus 1.749 to 3.471 g/L in control cultures. Despite reduced growth, high biohydrogen volumetric productivity was achieved, reaching 7.3 L/L·d, along with a significant yield of 2.642 mol of hydrogen per mole of glucose consumed. This represents 66.05% of the maximum stoichiometric yield calculated when acetic acid is the sole byproduct. Apparently, the presence of low concentrations of furfural and HMF released during the pre-treatment of Eichhornia crassipes not only negatively affects growth capacity but also diminishes butyric acid production, favoring biohydrogen production. © The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature 2024.