Open AccessA rapid method for purification of homogenous Legionella pneumophila cytotoxic protease using fast protein liquid chromatography
Author(s) -
Rechnitzer C.,
Tvede M.,
Döring G.
Publication year - 1989
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.1574-6968.1989.tb03079.x
Subject(s) - fast protein liquid chromatography , protease , legionella pneumophila , cytotoxic t cell , hela , microbiology and biotechnology , chromatography , biology , legionella , ultrafiltration (renal) , biochemistry , chemistry , bacteria , enzyme , in vitro , genetics
A purification method was developed to isolate Legionella pneumophila cytotoxic protease in a form suitable for biological assays. Culture supernatant of a clinical isolate of L. pneumophila , Knoxville 1 strain, was used as the starting material. The protease was purified by FPLC on a Mono Q column followed by ultrafiltration. The isolated proteolytic enzyme has a specific activity of 90 azocasein units/mg protein and is a 42 kDa monomeric protein as determined by SDS‐PAGE and gel filtration chromatography. It is heat‐labile and toxic to a variety of cells e.g. McCoy, SIRC, HeLa, and rhabdomyosarcoma cells, baby hamster and green monkey kidney cells, and human embryonic lung fibroblasts.