Open AccessPurification, crystallization and preliminary X‐ray analysis of a hexameric β‐glucosidase from wheat
Author(s) -
Sue Masayuki,
Yamazaki Kana,
Kouyama Junichi,
Sasaki Yasuyuki,
Ohsawa Kanju,
Miyamoto Toru,
Iwamura Hajime,
Yajima Shunsuke
Publication year - 2005
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309105027028
Subject(s) - crystallization , crystallography , monomer , aglycone , chemistry , substrate (aquarium) , solvent , escherichia coli , hepes , random hexamer , nuclear chemistry , stereochemistry , biochemistry , polymer , biology , organic chemistry , ecology , glycoside , gene
The wheat β‐glucosidase TaGlu1b, which is only active in a hexameric form, was tagged with 6×His at the N‐terminus, overexpressed in Escherichia coli and purified in two steps. The protein complexed with a substrate aglycone was crystallized at 293 K from a solution containing 10 m M HEPES pH 7.2, 1 M LiSO 4 and 150 m M NaCl using the hanging‐drop vapour‐diffusion method. Diffraction data were collected to 1.7 Å at the Photon Factory. The crystal belongs to space group P 4 1 32, with unit‐cell parameters a = b = c = 194.65 Å, α = β = γ = 90°. The asymmetric unit was confirmed by molecular‐replacement solution to contain one monomer, giving a solvent content of 72.1%.