Influence of Inhibitory Compounds on Biofuel Production from Oxalate-Rich Rhubarb Leaf Hydrolysates Using Thermoanaerobacter thermohydrosulfuricus Strain AK91

Abstract

The present investigation is on bioethanol and biohydrogen production from oxalate-rich rhubarb leaves which are an underutilized residue of rhubarb cultivation. Rhubarb leaves can be the feedstock for bioethanol and biohydrogen production using thermophilic, anaerobic bacteria. The fermentation of second-generation biomass to biofuels by Thermoanaerobacter has already been reported as well as their high ethanol and hydrogen yields although rhubarb biomass has not been examined for this purpose. Thermoanaerobacter thermohydrosulfuricus strain AK91 was characterized (temperature and pH optima, substrate utilization spectrum) which demonstrates that the strain can utilize most carbohydrates found in lignocellulosic biomass. Additionally, the influence of specific culture conditions, namely the partial pressure of hydrogen and initial glucose concentration, were investigated in batch culture and reveals that the strain is inhibited. Additionally, batch experiments containing common inhibitory compounds, namely carboxylic acids and aldehydes, some of which are present in high concentrations in rhubarb. Strain AK91 is not affected by alkanoic carboxylic acids and oxalate up to at least 100 mM although the strain was inhibited by 40 mM of malate. Interestingly, strain AK91 demonstrated the ability to reduce alkanoic carboxylic acids to their primary alcohols; more detailed studies with propionate as a model compound demonstrated that AK91’s growth is not severally impacted by high propionate loadings although 1-propanol titers did not exceed 8.5 mM. Additionally, ethanol and hydrogen production from grass and rhubarb leaf hydrolysates was investigated in batch culture for which AK91 produced 7.0 and 6.3 mM g−1, respectively.