Open AccessCrystallization and preliminary X‐ray analysis of pyridoxine 4‐oxidase, the first enzyme in pyridoxine degradation pathway I
Author(s) -
Mugo Andrew Njagi,
Kobayashi Jun,
Mikami Bunzo,
Ohnishi Kouhei,
Yagi Toshiharu
Publication year - 2012
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309111048688
Subject(s) - pyridoxine , enzyme , chemistry , pyridoxal , oxidase test , crystallization , pyridoxamine , pyridoxal phosphate , stereochemistry , biochemistry , crystallography , cofactor , organic chemistry
Vitamin B 6 ‐degradation pathway I has recently been identified in Mesorhizobium loti MAFF303099. Pyridoxine 4‐oxidase, an FAD‐dependent enzyme, is the first enzyme in this pathway and catalyzes the irreversible oxidation of pyridoxine to pyridoxal. The enzyme was overexpressed in Escherichia coli with a His 6 tag and purified. The recombinant enzyme was crystallized at 277 K by the sitting‐drop vapour‐diffusion method using PEG 4000 as the precipitant. The crystal, which belonged to space group P 2 1 2 1 2 1 with unit‐cell parameters a = 62.38, b = 79.44, c = 136.43 Å, diffracted to 2.2 Å resolution. The calculated V M value (3.19 Å 3 Da –1 ) suggested that the asymmetric unit contained one molecule.