<P><B>Abstract</B></P> <P>This work investigated the effect of ethylenediamine pretreatment on reducing enzyme loading in high gravity fermentation. At optimal conditions of ethylenediamine pretreatment, 85.5% lignin was removed. Enzyme adsorption analysis using a fluorescent cellulose-binding protein showed 35.2% increase of productive adsorption of enzymes to ethylenediamine pretreated biomass, which was caused by high delignification and dramatically increased surface roughness and porosity. In SScF at 15% glucan loading, up to 82.2 g/L ethanol was achieved with a relatively low enzyme loading of 3.6 FPU/g dry matter. It suggested that the remarkably high digestibility of EDA pretreated corn stover could effectively reduce the enzyme loading in the high gravity fermentation of cellulosic ethanol.</P> <P><B>Highlights</B></P> <P> <UL> <LI> EDA pretreatment removed 85.5% lignin and 42.7% hemicellulose. </LI> <LI> Surface roughness and porosity dramatically increased by EDA pretreatment. </LI> <LI> Productive adsorption of enzymes to EP-CS significantly increased by 35.2%. </LI> <LI> 82.2 g/L ethanol was achieved at low enzyme loading of 3.6 FPU/g dry matter. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>