Recombinant expression and characterization of a Xylella fastidiosa cysteine protease differentially expressed in a nonpathogenic strain

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...

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Bibliographic Details
Authors: Nogaroto, Viviane, Tagliavini, Sandra A., Gianotti, Andreia, Mikawa, Angela, Barros, Nilana M. T. [UNIFESP], Puzer, Luciano [UNIFESP], Carmona, Adriana K. [UNIFESP], Costa, Paulo I., Henrique-Silva, Flavio
Format: article
Status:Published version
Publication Date:2006
Country:Brasil
Institution:Universidade Federal de São Paulo (UNIFESP)
Repository:Repositório Institucional da UNIFESP
Language:English
OAI Identifier:oai:repositorio.unifesp.br:11600/29057
Online Access:http://dx.doi.org/10.1111/j.1574-6968.2006.00348.x
http://repositorio.unifesp.br/handle/11600/29057
Access Level:Open access
Keyword:Xylella fastidiosa
citrus variegated chlorosis
phytopathogen
recombinant expression
cysteine protease
xylellain
Description
Summary: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.

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