Open AccessExpression, purification, crystallization and preliminary X‐ray diffraction analysis of EtFPOX from Eupenicillium terrenum sp.
Author(s) -
Xing Keke,
Gan Weiqiong,
Jia Minze,
Gao Feng,
Gong Weimin
Publication year - 2013
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309113012128
Subject(s) - d amino acid oxidase , hydrogen peroxide , polyethylene glycol , chemistry , size exclusion chromatography , crystallography , crystallization , solvent , amino acid , x ray , molecule , escherichia coli , oxidase test , nuclear chemistry , enzyme , organic chemistry , biochemistry , physics , quantum mechanics , gene
The flavoenzyme fructosyl peptide oxidase (FPOX) catalyses the oxidative deglycation of fructosyl amino acids or fructosyl dipeptides to produce amino acids, glucosone and hydrogen peroxide. In this study, FPOX protein from Eupenicillium terrenum sp. (EtFPOX) was expressed in Escherichia coli and purified by Ni‐affinity and gel‐filtration chromatography. EtFPOX crystals were obtained using the sitting‐drop vapour‐diffusion method with polyethylene glycol 3350 as precipitant. X‐ray diffraction data were collected to 1.90 Å resolution using a synchrotron‐radiation source. The crystals belonged to space group P 2 1 2 1 2 1 , with unit‐cell parameters a = 65.6, b = 80.0, c = 83.4 Å, and contained one molecule in the asymmetric unit. The calculated Matthews coefficient and solvent content were 2.22 Å 3 Da −1 and 44.62%, respectively.