Cloning, expression, purification, crystallization and preliminary X-ray 
analysis of EaLsc, a levansucrase from Erwinia amylovora

Caputi, Lorenzo; Cianci, Michele; Benini, Stefano

doi:10.1107/S1744309113010750

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Cloning, expression, purification, crystallization and preliminary X-ray analysis of EaLsc, a levansucrase from Erwinia amylovora

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Caputi, L., Cianci, M., & Benini, S. (2013). Cloning, expression, purification, crystallization and preliminary X-ray analysis of EaLsc, a levansucrase from Erwinia amylovora. Acta Crystallographica Section F: Structural Biology and Crystallization Communications, F69, 570-573. doi:10.1107/S1744309113010750.

Cite as: https://hdl.handle.net/21.11116/0000-0009-C833-C

Abstract

The Gram-negative bacterium Erwinia amylovora is a destructive pathogen of Rosaceae. During infection, E. amylovora 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 E. amylovora (EaLsc) was heterologously expressed as a GST-fusion protein in Escherichia coli, purified and crystallized after tag removal. The protein crystallized in space group P2(1)2(1)2. X-ray diffraction data were acquired to 2.77 angstrom 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 angstrom(3) Da(-1).