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To elucidate the effects of lignin repolymerization on the combined pretreatment, the sequential acid and deep eutectic solvent (DES) pretreatment was implemented on Populus (poplar) in this study. Effects of acid pretreatment with or without phenolic additives (2-naphthol (2 N) and resorcinol (RS)) on the DES post-treatment efficiency were examined. The results showed that 2 N inhibited the lignin repolymerization in the first-step pretreatment, and slightly enhanced the lignin removal during DES post-treatment. However, RS intensified the lignin repolymerization, significantly decreasing the delignification of DES post-treatment from 73.6%∼79.4% to 39.0∼43.0%. Moreover, the poorer enzymatic hydrolysis was observed in the combined pretreated substrates as compared to the single DES pretreated ones, due to the higher surface lignin coverage. The addition of 2 N counteracted the adverse effects of acid pretreatment on the hydrolysis yields of combined pretreated substrates, and facilitated the yields from 25.8%∼36.0% to 26.2∼67.3%, while the RS addition further decreased the hydrolysis yields to 8.6∼9.6%. The mechanism for the changes on enzymatic digestibilities of the combined pretreated substrates was elucidated. [Display omitted] • 2-Naphthol inhibited lignin polymerization, while resorcinol intensified it. • Lignin repolymerization hindered the delignification of the DES post-treatment. • 2-Naphthol promoted the hydrolysis of pretreated poplar, but resorcinol impeded it. • 2-Naphthol addition benefited the lignin fractionation and valorization. [ABSTRACT FROM AUTHOR]

Highlights • Alkali and acid pretreated biomass slurries were mixed for ethanol production. • This approach saved 40–50% alkali/acid use. • This approach improved fermentability. Abstract Alkali and acid pretreatment based processes for lignocellulosic biofuel production typically consume substantial amount of alkali and acid. In addition, alkali pretreatment generates compounds with high toxicity rendering washing/detoxification inevitable. In this work, alkali (AL) and dilute acid (DA) pretreatment based processes were compared side by side for ethanol production from corn stover (CS) using commercial enzymes and Zymomonas mobilis 8b strain. Next, AL and DA pretreated CS slurries (without removing liquid stream) were mixed to neutralize for enzymatic hydrolysis and ethanol fermentation. The digestibility and fermentability of the mixed pretreated biomass were systematically studied and compared to unmixed pretreated biomass. Results showed that mixed biomass process saved 40–50% alkali/acid use and enhanced fermentability compared to AL-CS. This work suggests that it could be beneficial to have both alkali and acid pretreatments operating in a biorefinery, which could accommodate various feedstocks. [ABSTRACT FROM AUTHOR]