title: PelN is a new pectate lyase of Dickeya dadantii with unusual characteristics.
abstract: The plant-pathogenic bacterium Dickeya dadantii produces several pectinolytic enzymes that play a major role in the soft-rot disease. Eight characterized endopectate lyases are secreted in the extracellular medium by the type II secretion system, Out. They cleave internal glycosidic bonds of pectin, leading to plant tissue maceration. The D. dadantii pectate lyases belong to different families, namely, PL1, PL2, PL3, and PL9. Analysis of the D. dadantii 3937 genome revealed a gene encoding a new protein of the PL9 family, which already includes the secreted endopectate lyase PelL and the periplasmic exopectate lyase PelX. We demonstrated that PelN is an additional extracellular protein secreted by the Out system. However, PelN has some unusual characteristics. Although most pectate lyases require a very alkaline pH and Ca²⁺ for their activity, the PelN activity is optimal at pH 7.4 and in the presence of Fe²⁺ as a cofactor. PelN is only weakly affected by the degree of pectin methyl esterification. The PelN structural model, constructed on the basis of the PelL structure, suggests that the PelL global topology and its catalytic amino acids are conserved in PelN. Notable differences concern the presence of additional loops at the PelN surface, and the replacement of PelL charged residues, involved in substrate binding, by aromatic residues in PelN. The pelN expression is affected by different environmental conditions, such as pH, osmolarity, and temperature. It is controlled by the repressors KdgR and PecS and by the activator GacA, three regulators of D. dadantii pectinase genes. Since a pelN mutant had reduced virulence on chicory leaves, the PelN enzyme plays a role in plant infection, despite its low specific activity and its unusual cofactor requirement.
authors.affiliation: Université de Lyon, INSA-Lyon, MAP, CNRS UMR5240, Domaine Scientifique de la Doua, Villeurbanne, France.
authors.forename: Susan
authors.initials: S
authors.lastname: Hassan
authors.forename: Vladimir E
authors.initials: VE
authors.lastname: Shevchik
authors.forename: Xavier
authors.initials: X
authors.lastname: Robert
authors.forename: Nicole
authors.initials: N
authors.lastname: Hugouvieux-Cotte-Pattat
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015153:Blotting, Western; D004755:Enterobacteriaceae / Q000201:enzymology* / Q000472:pathogenicity; D008969:Molecular Sequence Data; D011133:Polysaccharide-Lyases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D011198:Solanum tuberosum / Q000382:microbiology
chemical_list: D001426:Bacterial Proteins; D011133:Polysaccharide-Lyases; C023911:pectate lyase