Crystallization and Preliminary X-Ray Diffraction Analysis of Glutaryl-7-aminocephalosporanic Acid Acylase from Pseudomonas sp. GK16

• Published in: Journal of structural biology Vol. 131; no. 1; pp. 79 - 81
• Main Authors: Kwon, Taek H., Rhee, Sangkee, Lee, Young S., Park, Sung S., Kim, Kyung H.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.07.2000
• Subjects: Amidohydrolases - chemistry; Amidohydrolases - genetics; Crystallization; Crystallography, X-Ray; glutaryl-7-aminocephalosporanic acid acylase; Mutation; Penicillin Amidase; Protein Conformation; protein crystallization; Protein Precursors - chemistry; Protein Precursors - genetics; Protein Processing, Post-Translational; Pseudomonas - enzymology; Pseudomonas sp. GK16; X-ray diffraction; X-ray diffraction; protein crystallization; Pseudomonas sp. GK16; glutaryl-7-aminocephalosporanic acid acylase
• ISSN: 1047-8477; 1095-8657
• DOI: 10.1006/jsbi.2000.4256

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 (αβ)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 Å resolution. The crystal is tetragonal with the space group P41212 or P43212 and the unit cell parameters are a = b = 73.5 Å, c = 380.3 Å. Considerations of the possible values of Vm account for the presence of a tetramer in the asymmetric unit.