Open AccessExpression, purification, crystallization and preliminary X‐ray diffraction analysis of Bifidobacterium adolescentis xylose isomerase
Author(s) -
dos Reis Caio Vinicius,
Bernardes Amanda,
Polikarpov Igor
Publication year - 2013
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s174430911301110x
Subject(s) - xylose isomerase , isomerase , xylose , chemistry , biochemistry , escherichia coli , polyethylene glycol , recombinant dna , enzyme , fermentation , gene
Xylose isomerase (EC 5.3.1.5) is a key enzyme in xylose metabolism which is industrially important for the transformation of glucose and xylose into fructose and xylulose, respectively. The Bifidobacterium adolescentis xylA gene (NC_008618.1) encoding xylose isomerase (XI) was cloned and the enzyme was overexpressed in Escherichia coli . Purified recombinant XI was crystallized using the sitting‐drop vapour‐diffusion method with polyethylene glycol 3350 as the precipitating agent. A complete native data set was collected to 1.7 Å resolution using a synchrotron‐radiation source. The crystals belonged to the orthorhombic space group P 2 1 2 1 2, with unit‐cell parameters a = 88.78, b = 123.98, c = 78.63 Å.