The samples were taken regularly for conductivity analysis using a DDS-307A conductivity meter (Shanghai INESA and Scientific Instrument Co., Shanghai, China), and sugars and inhibitors analysis on HPLC. The stover sugar hydrolysate was concentrated to a 300–350 g/L sugar concentration

by steam evaporation before hydrogenolysis. Then the concentrated stover sugar hydrolysate was sent to the hydrogenolysis C59 wnt cost reactor supplemented with 4% (w/w) sodium hydroxide and 15% modified Raney nickel catalyst #12-2 (w/w, based on the total sugar weight in system). The purified hydrogen was ventilated into the reactor to remove the inert air in the reactor and heated to 230 °C and 11.0 MPa slowly in an oil bath, then maintained for 120 min until glucose and signaling pathway xylose were completely converted. After each batch reaction, the Raney nickel catalyst was recycled by washing with deionized water then sent to the next round of catalytic operation. Glucose, xylose, inhibitory compounds, such as formic acid, furfural, 5-hydroxymethylfurfural (HMF), acetic acid and levulinic acid, and hydrogenolysis products, including ethanediol, 1,2-propanediol, butanediol, glycerol, sorbitol, lactic acid were determined using high-performance liquid chromatography (LC-20AD, refractive index detector RID-10A, Shimadzu, Japan) with a Bio-Rad Aminex

HPX-87H column at the column temperature of 65 °C. The mobile phase was 0.005 M H2SO4 at the rate of 0.6 mL/min. All the samples were diluted properly and filtered through a 0.22 μm filter before analysis. The protein content in the hydrolysate at different purification stages was determined according to Bradford using bovine serum albumin Fossariinae (BSA) for making

standard protein curve [17]. All the assays were performed in triplicates and the average data were presented. The compositions of virgin corn stover were analyzed using ANKOM 200 Cellulose Analyzer (ANKOM Technology, Macedon, NY, USA) [14]. The original corn stover contained 45.09 ± 0.08% glucan, 31.74 ± 0.18% xylan, 5.15 ± 0.34% acid-insoluble lignin, and 4.98 ± 0.28% ash. All the above data were calculated on the dry solid matter. The glucose and xylose yields were calculated using the following equations [18]: Glucoseyield(%)=[Glu]×Vf×[Biomass]×m×1.111×100% Xyloseyield(%)=[Xyl]×Vh×[Biomass]×m×1.136×100%where [Glu] and [Xyl] were the glucose and xylose concentration at the end of the hydrolysis (g/L), respectively; V was the final liquid volume of the hydrolysis system (L); f was the cellulose content in corn stover (g/g); h was the hemicellulose content in corn stover (g/g); [Biomass] was the solids loading of corn stover in the enzymatic hydrolysis system (%, w/w); m was the total weight of the hydrolysis system (g).