MOESM1 of Performance of three delignifying pretreatments on hardwoods: hydrolysis yields, comprehensive mass balances, and lignin properties

Additional file 1: Table S1. Composition and mass yields of hybrid poplar and eucalyptus prior to and following pretreatment. Cu-AHP: Cu-catalyzed alkaline hydrogen peroxide pretreatment; [Ch][Lys]: cholinium lysinate pretreatment; CELF: co-solvent enhanced lignocellulosic fractionation. Table S2. Assignments of signals observed in 13C/1H 2D (HSQC) NMR spectra of lignins. Figure S1. Enzymatic hydrolysis xylose yields for pretreated solids of hybrid poplar (A and B) and eucalyptus (C and D) prepared by Cu-AHP, CELF, and [Ch][Lys] pretreatments as a function of enzyme loading (mg protein/g glucan in pretreated solids) and hydrolysis time. Hydrolysis was performed at a 10% (wt/vol) solids loading with the pH buffered at 5.0 for 24 or 72 h. Figure S2. Size-exclusion chromatography (SEC) elution profiles and estimated values for number average molar mass ( $$ \bar{M}_{\text{N}} $$ M ¯ N ), weight average molar mass ( $$ \bar{M}_{\text{W}} $$ M ¯ W ), and polydispersity index (PDI) for (A) eucalyptus and (B) hybrid poplar. SEC was performed using a Waters UltrahydrogelTM 250 column with a mobile phase comprising a 80:20 (v/v) solution of 0.1 M NaNO3:0.005 M NaOH/CH3CN. Figure S3. Summary of the fate of the (A, D) glucan, (B, E) xylan, and (C, F) lignin for the three pretreatments for (A, B, C) hybrid poplar and (D, E, F) eucalyptus. Hydrolysis yields are for 30 mg/g glucan enzyme loading for 72 h. Figure S4. 13C/1H 2D (HSQC) NMR spectra of (a) native cellulolytic eucalyptus lignin, (b) eucalyptus 1st-stage Cu-AHP lignin, (c) eucalyptus 2nd-stage Cu-AHP lignin, (d) eucalyptus CELF lignin, (e) eucalyptus [Ch][Lys] lignin, (f) native cellulolytic poplar lignin, (g) poplar 1st-stage Cu-AHP lignin, (h) poplar 2nd-stage Cu-AHP lignin, (i) poplar CELF lignin, and (j) poplar [Ch][Lys] lignin. All NMR spectra were recorded in DMSO-d6 solvent and signals at δ = 2.5 ppm for 1H NMR and δ = 39.50 ppm for 13C NMR were considered as reference peaks to assign the 2D (HSQC) NMR signals. Figure S5. Comparison of S/G ratios as determined by quantitative thioacidolysis versus (semi-quantitative) 2-D HSQC NMR.