title: Expression, purification, crystallization and preliminary X-ray diffraction analysis of the pectin methylesterase from the sugar cane weevil Sphenophorus levis.
abstract: Pectin methylesterase removes the methyl groups from the main chain of pectin, the major component of the middle lamella of the plant cell wall. The enzyme is involved in plant cell-wall development, is part of the enzymatic arsenal used by microorganisms to attack plants and also has a wide range of applications in the industrial sector. Therefore, there is a considerable interest in studies of the structure and function of this enzyme. In this work, the pectin methylesterase from Sphenophorus levis was produced in Pichia pastoris and purified. Crystals belonging to the monoclinic space group C2, with unit-cell parameters a = 122.181, b = 82.213, c = 41.176 Å, β = 97.48°, were obtained by the sitting-drop vapour-diffusion method and an X-ray diffraction data set was collected to 2.1 Å resolution. Structure refinement and model building are in progress.
authors.affiliation: Instituto de Física de São Carlos, Universidade de São Paulo, Avenida Trabalhador Sãocarlense 400, 13566-590 São Carlos-SP, Brazil.
authors.forename: Danilo Elton
authors.initials: DE
authors.lastname: Evangelista
authors.affiliation: Instituto de Física de São Carlos, Universidade de São Paulo, Avenida Trabalhador Sãocarlense 400, 13566-590 São Carlos-SP, Brazil.
authors.forename: Andre
authors.initials: A
authors.lastname: Schutzer de Godoy
authors.affiliation: Departamento de Genética e Evolução, Universidade Federal de São Carlos, Rodovia Washington Luís, Km 235, 13565-905 São Carlos-SP, Brazil.
authors.forename: Fernando
authors.initials: F
authors.lastname: Fonseca Pereira de Paula
authors.affiliation: Departamento de Genética e Evolução, Universidade Federal de São Carlos, Rodovia Washington Luís, Km 235, 13565-905 São Carlos-SP, Brazil.
authors.forename: Flavio
authors.initials: F
authors.lastname: Henrique-Silva
authors.affiliation: Instituto de Física de São Carlos, Universidade de São Paulo, Avenida Trabalhador Sãocarlense 400, 13566-590 São Carlos-SP, Brazil.
authors.forename: Igor
authors.initials: I
authors.lastname: Polikarpov
mesh_terms: D000818:Animals; D002265:Carboxylic Ester Hydrolases / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D003460:Crystallization; D018360:Crystallography, X-Ray; D004926:Escherichia coli; D019476:Insect Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D048889:Weevils / Q000201:enzymology*
chemical_list: D019476:Insect Proteins; D002265:Carboxylic Ester Hydrolases; C022323:pectinesterase
keywords: Sphenophorus levis; pectin methylesterase