Open AccessCrystallization and X‐ray diffraction analysis of nylon hydrolase (NylC) from Arthrobacter sp. KI72
Author(s) -
Nagai Keisuke,
Yasuhira Kengo,
Tanaka Yusuke,
Kato Daiichiro,
Takeo Masahiro,
Higuchi Yoshiki,
Negoro Seiji,
Shibata Naoki
Publication year - 2013
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309113024263
Subject(s) - crystallization , orthorhombic crystal system , ammonium sulfate , column chromatography , chemistry , size exclusion chromatography , chromatography , arthrobacter , crystal (programming language) , crystallography , nuclear chemistry , crystal structure , organic chemistry , enzyme , computer science , programming language
Nylon hydrolase (NylC) encoded by Arthrobacter plasmid pOAD2 (NylC p2 ) was expressed in Escherichia coli JM109 and purified by ammonium sulfate fractionation, anion‐exchange column chromatography and gel‐filtration chromatography. NylC p2 was crystallized by the sitting‐drop vapour‐diffusion method with ammonium sulfate as a precipitant in 0.1 M HEPES buffer pH 7.5 containing 0.2 M NaCl and 25% glycerol. Diffraction data were collected from the native crystal to a resolution of 1.60 Å. The obtained crystal was spindle shaped and belonged to the C ‐centred orthorhombic space group C 222 1 , with unit‐cell parameters a = 70.84, b = 144.90, c = 129.05 Å. A rotation and translation search gave one clear solution containing two molecules per asymmetric unit.