Cloning, expression, purification, crystallization and preliminary X‐ray analysis of EaLsc, a levansucrase from Erwinia amylovora. Issue 5 (22nd May 2013)

• Record Type: Journal Article
• Authors: Caputi, Lorenzo; Cianci, Michele; Benini, Stefano
• Is Part Of: Acta crystallographica. Volume 69:Issue 5(2013:May)
• Page Start: 570
• Page End: 573
• Publication Date: 2013-05-22
• Subjects: Crystallography -- Periodicals; Crystals -- Periodicals; 548
• DOI: 10.1107/S1744309113010750
• Languages: English
• ISSNs: 1744-3091

Abstract

<abstract abstract-type="main" xml:lang="en"> <title> <x xml:space="preserve">Abstract</x> </title> <p>The Gram‐negative bacterium <italic>Erwinia amylovora</italic> is a destructive pathogen of Rosaceae. During infection, <italic>E. amylovora</italic> produces the exopolysaccharide levan, which contributes to the occlusion of plant vessels, causing the wilting of shoots. Levan is a fructose polymer that is synthesized by multifunctional enzymes called levansucrases. The levansucrase from <italic>E. amylovora</italic> (<italic>Ea</italic>Lsc) was heterologously expressed as a GST‐fusion protein in <italic>Escherichia coli</italic>, purified and crystallized after tag removal. The protein crystallized in space group <italic>P</italic>2₁2₁2. X‐ray diffraction data were acquired to 2.77 Å resolution. The structure of the enzyme was solved by molecular replacement. The asymmetric unit contains eight enzyme molecules, giving a solvent content of 58.74% and a Matthews coefficient of 2.98 ų Da^‐1.</p> </abstract>