Figure 1
TLC analysis of the products of RBcel1_WT activity on ClPNP β-G3 over a range of pH values from 4 to 9. 10 µM enzyme was incubated with 2 mM ClPNP β-G3 in 20 mM citrate–phosphate–CHES buffer for 5 min at 25°C. The following cellooligosaccharides and ClPNP derivatives were used as standards (1 mM of each; lane ML): glucose (G1), cellobiose (G2), cellotriose (G3), cellotetraose (G4), cellopentaose (G5), 4-nitrophenyl β-glucoside (PNP β-G1), 2-chloro-4-nitrophenyl β-cellobioside (ClPNP β-G2) and 2-chloro-4-nitrophenyl β-cellotrioside (ClPNP β-G3). The presence of 2-chloro-4-nitrophenyl β-cellotetraoside is marked by an arrow. |