Crystallization and preliminary X-ray diffraction analysis of the fructofuranosidase from Schwanniomyces occidentalis

Schwanniomyces occidentalis invertase is an extracellular enzyme that releases -fructose from the nonreducing termini of various -d-fructofuranoside substrates. Its ability to produce 6-kestose by transglycosylation makes this enzyme an interesting research target for applications in industrial biot...

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Detalles Bibliográficos
Autores: Polo, Aitana, Álvaro-Benito, Miguel, Fernández Lobato, María, Sanz-Aparicio, J.
Tipo de recurso: artículo
Fecha de publicación:2010
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/24450
Acceso en línea:http://hdl.handle.net/10261/24450
Access Level:acceso abierto
Palabra clave:Schwanniomyces occidentalis
X-ray crystallographic analyses
Descripción
Sumario:Schwanniomyces occidentalis invertase is an extracellular enzyme that releases -fructose from the nonreducing termini of various -d-fructofuranoside substrates. Its ability to produce 6-kestose by transglycosylation makes this enzyme an interesting research target for applications in industrial biotechnology. The enzyme has been expressed in Saccharomyces cerevisiae. Recombinant and wildtype forms, which showed different glycosylation patterns, were crystallized by vapour-diffusion methods. Although crystallization trials were conducted on both forms of the protein, crystals suitable for X-ray crystallographic analyses were only obtained from the wild-type enzyme. The crystals belonged to space group P212121, with unit-cell parameters a = 105.78, b = 119.49, c = 137.68 A ° . A diffraction data set was collected using a synchrotron source. Self-rotation function and sedimentation-velocity experiments suggested that the enzyme was dimeric with twofold symmetry.