title: Characterization of endoglucanase A from Clostridium cellulolyticum.
abstract: A construction was carried out to obtain a high level of expression in Escherichia coli of the gene celCCA, coding for the endoglucanase A from Clostridium cellulolyticum (EGCCA). The enzyme was purified in two forms with different molecular weights, 51,000 and 44,000. The smaller protein was probably the result of proteolysis, although great care was taken to prevent this process from occurring. Evidence was found for the loss of the conserved reiterated domains which are characteristic of C. thermocellum and C. cellulolyticum cellulases. The two forms were extensively studied, and it was demonstrated that although they had the same pH and temperature optima, they differed in their catalytic properties. The truncated protein gave the more efficient catalytic parameters on carboxymethyl cellulose and showed improved endoglucanase characteristics, whereas the intact enzyme showed truer cellulase characteristics. The possible role of clostridial reiterated domains in the hydrolytic activity toward crystalline cellulose is discussed.
authors.affiliation: Laboratoire de Chimie Bactérienne, Marseille, France.
authors.forename: H P
authors.initials: HP
authors.lastname: Fierobe
authors.forename: C
authors.initials: C
authors.lastname: Gaudin
authors.forename: A
authors.initials: A
authors.lastname: Belaich
authors.forename: M
authors.initials: M
authors.lastname: Loutfi
authors.forename: E
authors.initials: E
authors.lastname: Faure
authors.forename: C
authors.initials: C
authors.lastname: Bagnara
authors.forename: D
authors.initials: D
authors.lastname: Baty
authors.forename: J P
authors.initials: JP
authors.lastname: Belaich
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D002480:Cellulase / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D002482:Cellulose / Q000378:metabolism; D003001:Cloning, Molecular; D003013:Clostridium / Q000201:enzymology / Q000235:genetics*; D004269:DNA, Bacterial; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D004926:Escherichia coli / Q000235:genetics; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008969:Molecular Sequence Data; D010957:Plasmids; D013696:Temperature
chemical_list: D004269:DNA, Bacterial; D002482:Cellulose; D002480:Cellulase