Open AccessPurification, crystallization and preliminary X‐ray analysis of l ‐sorbose reductase from Gluconobacter frateurii complexed with l ‐sorbose or NADPH
Author(s) -
Kubota Keiko,
Nagata Koji,
Miyazono Kenichi,
Toyama Hirohide,
Matsushita Kazunobu,
Tanokura Masaru
Publication year - 2009
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309109014687
Subject(s) - sorbose , chemistry , crystallization , reductase , stereochemistry , enzyme , biochemistry , fructose , organic chemistry
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– l ‐sorbose complex and the SR–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– l ‐sorbose complex belonged to space group C 222 1 , with unit‐cell parameters a = 124.2, b = 124.1, c = 60.8 Å. The crystal of the SR–NADPH complex belonged to space group P 2 1 , with unit‐cell parameters a = 124.3, b = 61.0, c = 124.5 Å, β = 89.99°. The crystals contained two and eight molecules, respectively, in the asymmetric unit.