Endoglucanase Z from the phytopathogenic bacterium Erwinia chrysanthemi (strain 3937) was purified by affinity chromatography on microcrystalline cellulose Avicel PH101. A kinetic characterization using p-nitrophenyl β-d-cellobioside and p-nitrophenyl β-d-lactoside as substrates was conducted: endoglucanase Z exhibited Km values of 3 mM and 7.5 mM and Vm values of 129 and 40 nmol·min-1·mg-1 towards p-nitrophenyl β-d-cellobioside (kcat=0.1 s-1) and p-nitrophenyl β-d-lactoside (kcat=0.03 s-1), respectively). The hydrolysis of cellotetraitol by endoglucanase Z was followed by HPLC and 1H NMR. Results show that cellobiitol and β-cellobiose are initially formed, demonstrating that the enzyme is acting by a molecular mechanism retaining the anomeric configuration. This suggests the involvement of a glycosyl-enzyme intermediate. © 1992.
Barras, F., Bortoli-German, I., Bauzan, M., Rouvier, J., Gey, C., Heyraud, A., & Henrissat, B. (1992). Stereochemistry of the hydrolysis reaction catalyzed by endoglucanase Z from Erwinia chrysanthemi. FEBS Letters, 300(2), 145–148. https://doi.org/10.1016/0014-5793(92)80183-H