Open AccessCrystallization and preliminary X‐ray study of the deaminase AmnE from Pseudomonas sp. AP‐3
Author(s) -
Yu Dan,
Jiang Yongji,
Hou Jianfeng,
Chen Shuai,
Zhang Guofang,
Liu Xiang,
Dong Hui,
Yu Bo
Publication year - 2013
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309113016709
Subject(s) - orthorhombic crystal system , escherichia coli , crystallization , resolution (logic) , pseudomonas , chemistry , gene , microbiology and biotechnology , biochemistry , biology , crystallography , crystal structure , bacteria , organic chemistry , genetics , artificial intelligence , computer science
The amnE gene from Pseudomonas sp. AP‐3 has been verified as encoding a deaminase with 142 amino‐acid residues. In order to change the substrate specificity via structure‐based protein engineering, the amnE gene, after gene‐code optimization, was chemically synthesized and cloned into the expression vector pET‐28a. The protein was expressed in Escherichia coli BL21 (DE3) and purified by Ni 2+ ‐chelating affinity chromatography. Diffraction‐quality crystals were obtained using the hanging‐drop vapour‐diffusion method and diffracted to a resolution of 2.09 Å. The crystals belonged to the orthorhombic space group C 222 1 , with unit‐cell parameters a = 63.23, b = 88.93, c = 137.83 Å.