Purification, crystallization and preliminary X-ray analysis of l-sorbose reductase from Gluconobacter frateurii complexed with l-sorbose or NADPH
July 1st, 2009 Posted by: admin
NADPH-dependent l-sorbose reductase (SR) from Gluconobacter frateurii was expressed in Escherichia coli, purified and crystallized with l-sorbose or NADPH using the sitting-drop vapour-diffusion method at 293 K. Crystals of the SR[ndash]l-sorbose complex and the SR[ndash]NADPH complex were obtained using reservoir solutions containing PEG 2000 or PEG 400 as precipitants and diffracted X-rays to 2.38 and 1.90 Å resolution, respectively. The crystal of the SR[ndash]l-sorbose complex belonged to space group C2221, with unit-cell parameters a = 124.2, b = 124.1, c = 60.8 Å. The crystal of the SR[ndash]NADPH complex belonged to space group P21, with unit-cell parameters a = 124.3, b = 61.0, c = 124.5 Å, [beta] = 89.99°. The crystals contained two and eight molecules, respectively, in the asymmetric unit.
