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

ABSTRACT: Nylon hydrolase (NylC) encoded by Arthrobacter plasmid pOAD2 (NylCp2) was expressed in Escherichia coli JM109 and purified by ammonium sulfate fractionation, anion-exchange column chromatography and gel-filtration chromatography. NylCp2 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.

SUBMITTER: Nagai K

PROVIDER: S-EPMC3792678 | biostudies-literature | 2013 Oct

REPOSITORIES: biostudies-literature

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

Nagai Keisuke K Yasuhira Kengo K Tanaka Yusuke Y Kato Dai-ichiro D Takeo Masahiro M Higuchi Yoshiki Y Negoro Seiji S Shibata Naoki N

Acta crystallographica. Section F, Structural biology and crystallization communications 20130928 Pt 10

Nylon hydrolase (NylC) encoded by Arthrobacter plasmid pOAD2 (NylCp2) was expressed in Escherichia coli JM109 and purified by ammonium sulfate fractionation, anion-exchange column chromatography and gel-filtration chromatography. NylCp2 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 obtaine ...[more]

PMID: 24100570

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