Exploring Ternary Deep Eutectic Solvent Pretreatment in a One-Pot Process with Napier Grass for Bioethanol Production

Effective pretreatment is essential for successfully utilizing renewable resources such as lignocellulosic biomass in the production of bioethanol. In this study, ternary deep eutectic solvents (DESs), namely choline chloride/lactic acid/glycerol (ChCl/LA/Gly), choline chloride/oxalic acid/glycerol (ChCl/OA/Gly), choline chloride/lactic acid/ethylene glycol (ChCl/LA/EG), and choline chloride/oxalic acid/ethylene glycol (ChCl/OA/EG) were prepared and employed for the pretreatment of cellulose-rich Napier grass (NG). Post treatment, the NG hydrolysate was subjected to enzymatic saccharification followed by ethanol fermentation. The results showed effective delignification of NG after treatment with the prepared ternary DESs, with ChCl/LA/EG removing a maximum of 92.89% lignin. The efficiency of the prepared DESs is attributed to their low densities, pH, and viscosity. Enzymatic saccharification of ChCl/LA/EG-treated NG resulted in a 1.68 fold increase in reducing sugar yield compared to that of untreated NG. All pretreated NG produced more bioethanol via a separate hydrolysis and fermentation (SHF) process than untreated NG after Saccharomyces cerevisiae fermentation. A maximum of 0.37 g bioethanol/g of biomass was obtained from the one-pot process using ChCl/LA/Gly pretreatment. FTIR and XRD analyses of untreated and pretreated NG corroborated the efficacy of the ternary DESs on cellulose recovery and delignification. Also, enzymatic and microbial inhibition studies on the prepared DESs show their potential to be employed in a one-pot process for biorefinery. The results of the present investigation show the potential of utilizing eco-friendly DESs and renewable resources for the production of bioethanol, a viable option to fossil fuels.