Open AccessProteolytic activity among various oral Treponema species and cloning of a prtP ‐like gene of Treponema socranskii subsp. socranskii 1
Author(s) -
Heuner Klaus,
Bergmann Ingo,
Heckenbach Kirsten,
Göbel Ulf B
Publication year - 2001
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2001.tb10752.x
Subject(s) - treponema , microbiology and biotechnology , proteases , chymotrypsin , biology , casein , gene , protease , cloning (programming) , molecular cloning , trypsin , biochemistry , chemistry , enzyme , peptide sequence , virology , syphilis , human immunodeficiency virus (hiv) , computer science , programming language
The proteolytic activity of 11 treponemal strains representing different phylogenetic groups was investigated by SDS–polyacrylamide gel electrophoresis with copolymerised casein, gelatin and fibrinogen as substrates. The activity was specified to be trypsin‐ and chymotrypsin‐like by the cleavage of synthetic substrates BAPNA and SAAPFNA, respectively. Nine strains degrade casein and the synthetic substrate BAPNA. Chymotrypsin‐like activity specifically inhibited by phenylmethylsulfonyl fluoride was found in four treponemes. Southern blot analysis using a Treponema socranskii subsp. socranskii ‐specific prtP probe confirmed the presence of prtP homologous genes in these four strains. The internal fragments of the chymotrypsin‐like protease genes were cloned and sequenced after PCR amplification. Here we report the cloning of the complete prtP ‐like gene of T. socranskii subsp. socranskii , an organism shown to possess epidemiologic relevance in periodontitis.