Overproduction, crystallization and preliminary X-ray analysis of the putative L-ascorbate-6-phosphate lactonase UlaG from Escherichia coli.
UlaG, the putative L-ascorbate-6-phosphate lactonase encoded by the ulaG gene from the utilization of L-ascorbate regulon in Escherichia coli, has been cloned, overexpressed, purified using standard chromatographic techniques and crystallized. Crystals were obtained by sitting-drop vapour diffusion at 293 K. Preliminary X-ray diffraction analysis revealed that the UlaG crystals belonged to the monoclinic space group C2, with unit-cell parameters a = 104.52, b = 180.69, c = 112.88 A, beta = 103.26 degrees. The asymmetric unit is expected to contain six copies of UlaG, with a corresponding volume per protein weight of 2.16 A3 Da(-1) and a solvent content of 43%.
Garces
Fernando
F
Structural Biology Department, Institut de Biologia Molecular de Barcelona and Institut de Recerca Biomèdica, E-08028 Barcelona, Spain.
Fernández
Francisco J
FJ
Pérez-Luque
Rosa
R
Aguilar
Juan
J
Baldomà
Laura
L
Coll
Miquel
M
Badía
Josefa
J
Vega
M Cristina
MC
D001483:Base Sequence; D002265:Carboxylic Ester Hydrolases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000201:enzymology*; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D014961:X-Ray Diffraction
D029968:Escherichia coli Proteins; D011994:Recombinant Proteins; D002265:Carboxylic Ester Hydrolases; C527649:L-ascorbate 6-phosphate lactonase, E coli; C019103:delta-lactonase