Journal
RESEARCH IN MICROBIOLOGY
Volume 155, Issue 1, Pages 31-38Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.resmic.2003.09.010
Keywords
Prevotella nigrescens; Prevotella intermedia; hemolysins
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Hemolytic activity was evaluated in the putative periodontopathogens Prevotella intermedia and Prevotella nigrescens. Whole cells of both species present weak hemolytic activity evidenced only by solid media assays after 48 h of bacterial growth or after 5 h of interaction with erythrocytes at 37 degreesC in liquid assays. In this work we show that the use of crude extract allowed the detection of a higher hemolytic activity for P. intermedia, but surprisingly not for P. nigrescens. Incubation at 37 degreesC for 9 h, or treatment with trypsin or proteinase K, increased or exposed the hemolytic activity of R intermedia and P. nigrescens crude extract, respectively. The activation process was inhibited by TLCK and PMSF but not by EDTA, E-64 or pepstatin A, indicating the serino-protease nature of the factor involved in activation of P intermedia and P. nigrescens hemolysins. Both the buffer and the pH employed for cell fractionation influenced the activation of hemolysin, and the best results were obtained with Universal buffer at pH 8.0. The activated hemolysins acted optimally at pH 6.5 at 37 degreesC and the maximum hemolytic activity was detected at the early log phase of growth. The results of this study show for the first time a strong hemolytic activity for P. nigrescens and evidence of proteolytic activation of hemolysins produced by periodontopathogens. (C) 2003 Elsevier SAS. All rights reserved.
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