Purification and partial characterization of two extracellular endoglucanases from Cellulomonas fermentans.
Avicelase assay of gel slices after non-denaturing polyacrylamide gel electrophoresis of concentrated supernatants from Cellulomonas fermentans revealed four active bands. One of them corresponded to the principal active band on CM-cellulose. Among the three others, at least one did not correspond to any active band on CM-cellulose and might reflect the presence of an exoglucanase (EC 3.2.1.91). The active band on CM-cellulose was composed of two endoglucanases (EC 3.2.1.4), called CFA and CFB, which we purified by the means of DEAE-Trisacryl chromatography and high performance liquid chromatography (anion exchange chromatography and gel chromatography). These two monomeric enzymes differ in their molecular weights (40,000 and 57,000 for CFA and CFB, respectively) and in their catalytic constants in the reaction with CM-cellulose (Km were 1.5 g/l and 59 g/l for CFA and CFB, respectively), but have similar modes of action on this substrate and similar substrate specificities.
D000192:Actinomycetales / Q000201:enzymology*; D002480:Cellulase / Q000032:analysis / Q000302:isolation & purification*; D002482:Cellulose / Q000378:metabolism; D006868:Hydrolysis; D007700:Kinetics; D008970:Molecular Weight; D013379:Substrate Specificity
D002482:Cellulose; D002480:Cellulase