Open AccessRecombinant expression and characterization of a Xylella fastidiosa cysteine protease differentially expressed in a nonpathogenic strain
Author(s) -
Nogaroto Viviane,
Tagliavini Sandra A.,
Gianotti Andréia,
Mikawa Angela ,
Barros Nilana M.T.,
Puzer Luciano,
Carmona Adriana K.,
Costa Paulo I.,
HenriqueSilva Flávio
Publication year - 2006
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.1574-6968.2006.00348.x
Subject(s) - xylella fastidiosa , recombinant dna , biology , microbiology and biotechnology , strain (injury) , bacteria , protease , cysteine protease , enzyme , biochemistry , gene , genetics , anatomy
Xylella fastidiosa is a xylem‐limited, Gram‐negative bacterium responsible for citrus variegated chlorosis (CVC) in sweet oranges. In the present study, we present the recombinant expression, purification and characterization of an X. fastidiosa cysteine protease (dubbed Xylellain). The recombinant Xylellain ( HIS Xylellain) was able to hydrolyze carbobenzoxy‐Phe‐Arg‐7‐amido‐4‐methylcoumarin (Z‐FR‐MCA) and carbobenzoxy‐Arg‐Arg‐7‐amido‐4‐methylcoumarin (Z‐RR‐MCA) with similar catalytic efficiencies, suggesting that this enzyme presents substrate specificity requirements similar to cathepsin B. The immunization of mice with HIS Xylellain provided us with antibodies, which recognized a protein of c . 31 kDa in the X. fastidiosa pathogenic strains 9a5c, and X. fastidiosa isolated from coffee plants. However, these antibodies recognized no protein in the nonpathogenic X. fastidiosa J1a12, suggesting the absence or low expression of this protein in the strain. These findings enabled us to identify Xylellain as a putative target for combating CVC and other diseases caused by X. fastidiosa strains.