Overexpression, crystallization and preliminary X-ray analysis of xylulose-5-phosphate/fructose-6-phosphate phosphoketolase from Bifidobacterium breve.
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 mM thiamine diphosphate, 0.25 mM MgCl2, 24%(w/v) PEG 6000 and 0.1 M Bicine pH 9.0. The crystals belonged to the tetragonal space group I422, with unit-cell parameters a=b=174.8, c=163.8 A, and diffracted to beyond 1.7 A resolution.
Suzuki
Ryuichiro
R
Department of Biotechnology, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
Kim
Byung-Jun
BJ
Shibata
Tsuyoshi
T
Iwamoto
Yuki
Y
Katayama
Takane
T
Ashida
Hisashi
H
Wakagi
Takayoshi
T
Shoun
Hirofumi
H
Fushinobu
Shinya
S
Yamamoto
Kenji
K
D000446:Aldehyde-Lyases / Q000737:chemistry* / Q000235:genetics; D001644:Bifidobacterium / Q000201:enzymology*; D003460:Crystallization; D018360:Crystallography, X-Ray; D015870:Gene Expression
D000446:Aldehyde-Lyases; C065556:fructose-6-phosphate phosphoketolase