title: Characterization of proteolytic activities of Fusobacterium nucleatum.
abstract: This investigation attempted to detect the proteolytic activity of Fusobacterium nucleatum in living cells, lysate cells, and supernatant of cultures. The reactions were optimized in their pH, temperature, reaction time, enzyme source, and substrate volume. Synthetic substrates beta-naphthylamides (Cys-Na, Ser-Na, Leu-Na, Glu-Na, Lys-Na and BANA), carbobenzoxy L-tirosine p-nitrophenylester (CTN), and natural substrate azoalbumin were used. Reaction occurred with Cys-Na, Ser-Na, and Glu-Na in living cells and with Glu-Na, Leu-Na, and CTN substrates in lysate cells. The supernatant reacted only with Glu-Na. Optimal pH ranged from 6.0 to 7.5, except for CTN (pH 13), and optimal temperature, between 30 and 40 degrees C. Optimal reaction time was 60 min, except for Glu-Na in living cells (40 min), lysate cells (20 min), and CTN substrate (80 min). There was no activity with Lys-Na, BANA, and azoalbumin. Proteolytic activity was assessed by several inhibitors and the presence of metallo, serine, cysteine, and aspartic proteases were detected.
authors.affiliation: Departamento de Microbiologia/CCB, Universidade Estadual de Londrina, Londrina Paraná, Brasil.
authors.forename: Alice Tayoko
authors.initials: AT
authors.lastname: Ogawa
authors.forename: Tatiana de Arruda Campos
authors.initials: Tde A
authors.lastname: Brasil de Souza
authors.forename: Milton
authors.initials: M
authors.lastname: de Uzeda
authors.forename: Jose V
authors.initials: JV
authors.lastname: Jankevicius
authors.forename: Shiduca Itow
authors.initials: SI
authors.lastname: Jankevicius
mesh_terms: D000818:Animals; D002417:Cattle; D016967:Fusobacterium nucleatum / Q000201:enzymology*; D010447:Peptide Hydrolases / Q000032:analysis / Q000187:drug effects; D010518:Periodontitis / Q000201:enzymology*; D011480:Protease Inhibitors / Q000378:metabolism / Q000494:pharmacology
chemical_list: D011480:Protease Inhibitors; D010447:Peptide Hydrolases