title: Isolation, heterologous expression and characterization of an endo-polygalacturonase produced by the phytopathogen Burkholderia cepacia.
abstract: Endo-polygalacturonases (endoPGs) belong to the glycoside hydrolase family 28 and hydrolyze the alpha-1,4 glycosidic bond present in the smooth regions of pectins. Pectic substances are among the principal macromolecular components of the primary plant cell walls and are subjected to enzymatic degradation not only in the course of important physiological processes such as plant senescence and ripening, but also during infection events by plant pathogens. Here we report, for the first time, the isolation and the purification of an endoPG (PehA) from the supernatant of the plant pathogen Burkholderia cepacia strain ATCC 25416. In order to obtain adequate amounts of protein required for structural and functional studies, the gene coding for pehA was PCR-amplified and cloned in Escherichia coli cells. The recombinant protein was purified to homogeneity and characterized. PehA exhibited a pI value of 8.0 and an optimal activity at pH 3.5. Far-UV circular dichroism (CD) measurements show that PehA assumes a beta-helix fold super-secondary structural motif.
authors.affiliation: International School for Advanced Studies, Trieste, Italy. massa@sissa.it <massa@sissa.it>
authors.forename: Claudia
authors.initials: C
authors.lastname: Massa
authors.forename: Giuliano
authors.initials: G
authors.lastname: Degrassi
authors.forename: Giulia
authors.initials: G
authors.lastname: Devescovi
authors.forename: Vittorio
authors.initials: V
authors.lastname: Venturi
authors.forename: Doriano
authors.initials: D
authors.lastname: Lamba
mesh_terms: D000595:Amino Acid Sequence; D016956:Burkholderia cepacia / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D004926:Escherichia coli / Q000378:metabolism; D008969:Molecular Sequence Data; D011096:Polygalacturonase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D011994:Recombinant Proteins / Q000302:isolation & purification
chemical_list: D011994:Recombinant Proteins; D011096:Polygalacturonase