title: Protease activity of Clostridium difficile strains.
abstract: The production of proteolytic enzymes by 10 Clostridium difficile isolates of varying toxigenicity and clinical origin was studied to determine if all isolates secreted proteases. Different protease substrates were studied: gelatin, collagen, phenylazobenzyloxycarbonyl-leucyl-glycyl-L-prolyl-D-arginine (Pz-peptide), casein, azocasein, and azocoll. All isolates degraded gelatin, collagen, and azocoll. The supernatants of all isolates contained an enzyme capable of attacking gelatin incorporated in a polyacrylamide gel (zymograms) and forming two closely spaced lytic bands with an estimated molecular mass of 35-40 kDa. Polyclonal antibodies, produced against the C. difficile gelatinase, revealed in Western blots a 35-kDa protein in the culture supernatants of all C. difficile isolates. In the same manner, Clostridium perfringens collagenase polyclonal antibodies detected a 120-kDa protein in the culture supernatants of all isolates; this suggests that at least two proteases may exist in C. difficile. The protease activities of the 10 strains examined did not seem strikingly different quantitatively but were in general weak and their role in pathogenicity is suspect.
authors.affiliation: Département de microbiologie, Faculté de pharmacie, Université Paris-Sud, Chatenay-Malabry, France.
authors.forename: I
authors.initials: I
authors.lastname: Poilane
authors.forename: T
authors.initials: T
authors.lastname: Karjalainen
authors.forename: M C
authors.initials: MC
authors.lastname: Barc
authors.forename: P
authors.initials: P
authors.lastname: Bourlioux
authors.forename: A
authors.initials: A
authors.lastname: Collignon
mesh_terms: D002364:Caseins / Q000378:metabolism; D016360:Clostridioides difficile / Q000201:enzymology* / Q000472:pathogenicity; D003094:Collagen / Q000378:metabolism; D010450:Endopeptidases / Q000378:metabolism*; D004761:Enterocolitis, Pseudomembranous / Q000382:microbiology; D005780:Gelatin / Q000378:metabolism; D006801:Humans
chemical_list: D002364:Caseins; D005780:Gelatin; D003094:Collagen; D010450:Endopeptidases