Crystallization and Preliminary X-Ray Diffraction Analysis of Glutaryl-7-aminocephalosporanic Acid Acylase from Pseudomonas sp. GK16
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 penici...
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Published in | Journal of structural biology Vol. 131; no. 1; pp. 79 - 81 |
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Main Authors | , , , , |
Format | Journal Article |
Language | English |
Published |
United States
Elsevier Inc
01.07.2000
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Subjects | |
Online Access | Get full text |
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Summary: | 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. |
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Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
ISSN: | 1047-8477 1095-8657 |
DOI: | 10.1006/jsbi.2000.4256 |