Expression, purification, crystallization and preliminary X-ray diffraction analysis of the pectin methylesterase from the sugar cane weevil Sphenophorus levis

• Published in: Acta crystallographica. Section F, Structural biology communications Vol. 70; no. 3; pp. 331 - 334
• Main Authors: Evangelista, Danilo Elton, Schutzer de Godoy, Andre, Fonseca Pereira de Paula, Fernando, Henrique-Silva, Flavio, Polikarpov, Igor
• Format: Journal Article
• Language: English
• Published: 5 Abbey Square, Chester, Cheshire CH1 2HU, England International Union of Crystallography 01-03-2014; Wiley Subscription Services, Inc
• Subjects: Animals; Carboxylic Ester Hydrolases - biosynthesis; Carboxylic Ester Hydrolases - chemistry; Carboxylic Ester Hydrolases - isolation & purification; Crystallization; Crystallization Communications; Crystallography, X-Ray; Escherichia coli; Insect Proteins - biosynthesis; Insect Proteins - chemistry; Insect Proteins - isolation & purification; pectin methylesterase; Sphenophorus levis; Weevils - enzymology; pectin methylesterase; Sphenophorus levis
• ISSN: 2053-230X; 2053-230X
• DOI: 10.1107/S2053230X14001630

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.