title: Crystallization and preliminary X-Ray diffraction analysis of glutaryl-7-aminocephalosporanic acid acylase from Pseudomonas sp. GK16.
abstract: Glutaryl-7-aminocephalosporanicacid acylase from Pseudomonas sp. GK16 produces glutaryl-7-aminocephalosporanic acid, a key intermediate for the synthesis of cephem antibiotics. Sequence alignment suggests that the enzyme may belong to the N-terminal nucleophile hydrolase superfamily including penicillin G acylase. The enzyme is an (alphabeta)(2) heterotetramer of two nonidentical subunits. These subunits are derived from a nascent precursor polypeptide that is cleaved proteolytically through a two-step autocatalytic process upon folding. The enzyme has been crystallized using the vapor diffusion method. A bipyramidal crystal form was obtained from a solution containing polyethylene glycol (MW 3350) and calcium chloride. Complete diffraction data sets have been collected up to 2.8 A resolution. The crystal is tetragonal with the space group P4(1)2(1)2 or P4(3)2(1)2 and the unit cell parameters are a = b = 73.5 A, c = 380.3 A. Considerations of the possible values of V(m) account for the presence of a tetramer in the asymmetric unit.
authors.affiliation: Graduate School of Biotechnology, Korea University, Seoul, 136-701, Korea.
authors.forename: T H
authors.initials: TH
authors.lastname: Kwon
authors.forename: S
authors.initials: S
authors.lastname: Rhee
authors.forename: Y S
authors.initials: YS
authors.lastname: Lee
authors.forename: S S
authors.initials: SS
authors.lastname: Park
authors.forename: K H
authors.initials: KH
authors.lastname: Kim
mesh_terms: D000581:Amidohydrolases / Q000737:chemistry* / Q000235:genetics; D003460:Crystallization; D018360:Crystallography, X-Ray; D009154:Mutation; D010399:Penicillin Amidase; D011487:Protein Conformation; D011498:Protein Precursors / Q000737:chemistry / Q000235:genetics; D011499:Protein Processing, Post-Translational; D011549:Pseudomonas / Q000201:enzymology*
chemical_list: D011498:Protein Precursors; D000581:Amidohydrolases; D010399:Penicillin Amidase; C046586:glutarylamidocephalosporanic acid acylase