Crystallization and preliminary X-ray diffraction studies on recombinant isopenicillin N synthase from Aspergillus nidulans.
Recombinant Aspergillus nidulans isopenicillin N synthase was purified from an Escherichia coli expression system. The apoenzyme in the presence of saturating concentrations of MnCl2 could be crystallized by either macro- or microseeding, using the hanging drop vapor diffusion technique with polyethylene glycol 8000 as precipitant. The crystals (0.5-1.0 mm overall dimensions) diffract X-rays to at least 2.0 A resolution at synchrotrons and belong to space group P212121 with unit cell dimensions of a = 59.2 A, b = 127.0 A, and c = 139.6 A. The asymmetric unit contains one dimer, and the solvent content of the crystals is 60%. The crystals are radiation sensitive.
Roach
P L
PL
Dyson Perrins Laboratory, University of Oxford, United Kingdom.
Schofield
C J
CJ
Baldwin
J E
JE
Clifton
I J
IJ
Hajdu
J
J
D001233:Aspergillus nidulans / Q000201:enzymology; D003460:Crystallization; D018360:Crystallography, X-Ray; D015394:Molecular Structure; D010088:Oxidoreductases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D011092:Polyethylene Glycols; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification
D011994:Recombinant Proteins; D011092:Polyethylene Glycols; D010088:Oxidoreductases; C038370:isopenicillin N synthetase