Crystallization and X-ray diffraction analysis of nylon hydrolase (NylC) from Arthrobacter sp. KI72

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 C222₁, 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.