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[Display omitted] • High proportion of di-rhamnolipid can enhance enzymatic hydrolysis. • Incorporation of di-rhamnolipid in enzymatic hydrolysis yielded 196 g/L glucose. • 89.1 g/L ethanol was obtained from the fermentation of enzymatic hydrolysate. High-solids enzymatic hydrolysis is the premise of obtaining high concentration ethanol by fermentation. In this study, corn stover was first pretreated with formic acid under mild conditions, and more than 70 % of xylan and lignin were removed within the first hour. 173.0 g/L glucose was achieved from total 30 % solid of the pretreated corn stover via fed-batch mode. Moreover, the glucose concentration rose to 194.5 g/L and the hydrolysis time was significantly reduced by 42.9 % with the addition of di-rhamnolipid. On this basis, 89.1 g/L ethanol was obtained by fermentation, and the presence of di-rhamnolipid had no negative effect on fermentation. The effective conversion of corn stover to high titer ethanol provides support for the conversion of stover to ethanol in industrial production. [ABSTRACT FROM AUTHOR]

[Display omitted] • An ordered pre-incubated strategy is key to the auxiliary effect of PEG in high-solids hydrolysis. • The viscosity in a high-solids system was greatly decreased by pre-incubating substrate with PEG. • PEG pre-incubated enzyme demonstrated the least lignin adsorption capacity. High-solids enzymatic hydrolysis for biomass has currently received considerable interest. However, the solid effect during the process limits its economic feasibility. This work presented an ordered polyethylene glycol (PEG) pre-incubated strategy for enhancing the auxiliary effect of PEG in a high-solids enzymatic hydrolysis system. The substrate and enzyme were separately pre-incubated with PEG in this strategy. The ordered PEG pre-incubated strategies yielded a maximum glucose concentration of 166.6 g/L from 32 % (w/v) pretreated corncob with an enzymatic yield of 94.1 % by 72 h hydrolysis. Using this method, PEG not only lessened the lignin adsorption to cellulase but also altered particle rheological characteristics in the high-solids enzymatic hydrolysis system as a viscosity modifier. This study offered a new insight into the mechanism behind the PEG synergistic effect and would make it possible to achieve efficient high-solids loading hydrolysis in the commercial manufacture of cellulosic ethanol. [ABSTRACT FROM AUTHOR]