Characterization of proteolytic activities of Fusobacterium nucleatum.
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.
Ogawa
Alice Tayoko
AT
Departamento de Microbiologia/CCB, Universidade Estadual de Londrina, Londrina Paraná, Brasil.
Brasil de Souza
Tatiana de Arruda Campos
Tde A
de Uzeda
Milton
M
Jankevicius
Jose V
JV
Jankevicius
Shiduca Itow
SI
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
D011480:Protease Inhibitors; D010447:Peptide Hydrolases