Open AccessOverexpression, crystallization and preliminary X‐ray analysis of xylulose‐5‐phosphate/fructose‐6‐phosphate phosphoketolase from Bifidobacterium breve
Author(s) -
Suzuki Ryuichiro,
Kim ByungJun,
Shibata Tsuyoshi,
Iwamoto Yuki,
Katayama Takane,
Ashida Hisashi,
Wakagi Takayoshi,
Shoun Hirofumi,
Fushinobu Shinya,
Yamamoto Kenji
Publication year - 2010
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309110023845
Subject(s) - escherichia coli , bifidobacterium breve , phosphate , chemistry , crystallization , thiamine , biochemistry , microbiology and biotechnology , biology , bifidobacterium , gene , fermentation , organic chemistry , lactobacillus
The xylulose‐5‐phosphate/fructose‐6‐phosphate phosphoketolase gene from Bifidobacterium breve was cloned and overexpressed in Escherichia coli . The enzyme was purified to homogeneity and crystallized by the sitting‐drop vapour‐diffusion method. Crystals were obtained at 293 K using 0.05 m M thiamine diphosphate, 0.25 m M MgCl 2 , 24%( w / v ) PEG 6000 and 0.1 M Bicine pH 9.0. The crystals belonged to the tetragonal space group I 422, with unit‐cell parameters a = b = 174.8, c = 163.8 Å, and diffracted to beyond 1.7 Å resolution.