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

摘要

Schwanniomyces occidentalis invertase is an extracellular enzyme that releases\ud -fructose from the nonreducing termini of various -d-fructofuranoside substrates.\udIts ability to produce 6-kestose by transglycosylation makes this enzyme\udan interesting research target for applications in industrial biotechnology. The\udenzyme has been expressed in Saccharomyces cerevisiae. Recombinant and wildtype\udforms, which showed different glycosylation patterns, were crystallized by\udvapour-diffusion methods. Although crystallization trials were conducted on\udboth forms of the protein, crystals suitable for X-ray crystallographic analyses\udwere only obtained from the wild-type enzyme. The crystals belonged to space\udgroup P212121, with unit-cell parameters a = 105.78, b = 119.49, c = 137.68 A ° . A\uddiffraction data set was collected using a synchrotron source. Self-rotation\udfunction and sedimentation-velocity experiments suggested that the enzyme was\uddimeric with twofold symmetry.