Figure 3

Capillary zone electrophoresis of APTS-labeled oligosaccharides and SDS-PAGE analysis of FPLC positive screened derived fraction. (A) Enzymatic activity mediated by IEX4-37 on cellopentaose, a positively screened FPLC fraction derived from C. gestroi crude extract with a unique peptide match to a GH7. The enzyme source specifically cleaved internal glycosidic bonds of APTS-reducing-end-labeled-cellopentaose (arrows), releasing mainly cellotriose as hydrolysis product. (B) Complete hydrolysis of APTS-reducing-end-labeled-mannohexaose by IEX1-26, a positively screened FPLC fraction derived from C. gestroi crude extract with a unique peptide match to a GH2. The enzyme source specifically cleaved internal glycosidic bonds of mannohexaose (arrows), releasing mainly mannobiose and mannotriose as final product. (C) SDS-PAGE analysis of fractions IEX4-37 (lane1) and IEX1-26 (lane 2). A unique band of 50 kDa was observed after Coomassie blue staining for IEX4-37 (lane 1), and bands of about 80 kDa were observed for IEX1-26 after silver staining. (M) Protein Ladder. APTS: 8-aminopyreno-1,3,6-trisulfonic acid; FPLC: fast performance liquid chromatography; GH: glycoside hydrolases; IEX: ion-exchange.