Crystallization and preliminary X-ray diffraction analysis of glutaryl 7-aminocephalosporanic acid acylase from Pseudomonas sp. GK16

Taek H. Kwon, Sangkee Rhee, Youngsik Lee, Sung Soo Park, Kyung H. Kim

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Glutaryl-7-aminocephalosporanic acid 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 A resolution. The crystal is tetragonal with the space group P41212 or P43212 and the unit cell parameters are a = b = 73.5 A, c = 380.3. Considerations of the possible values of V(m) account for the presence of a tetramer in the asymmetric unit. (C) 2000 Academic Press.

Original languageEnglish
Pages (from-to)79-81
Number of pages3
JournalJournal of Structural Biology
Volume131
Issue number1
DOIs
Publication statusPublished - 2000 Sep 25

Fingerprint

amidase
Crystallization
Pseudomonas
X-Ray Diffraction
Enzymes
Penicillin Amidase
Calcium Chloride
Sequence Alignment
Anti-Bacterial Agents
Peptides
glutaryl-7-aminocephalosporanic acid

Keywords

  • Glutaryl-7-aminocephalosporanic acid acylase
  • Protein crystallization
  • Pseudomonas sp. GK16
  • X-ray diffraction

ASJC Scopus subject areas

  • Structural Biology

Cite this

Crystallization and preliminary X-ray diffraction analysis of glutaryl 7-aminocephalosporanic acid acylase from Pseudomonas sp. GK16. / Kwon, Taek H.; Rhee, Sangkee; Lee, Youngsik; Park, Sung Soo; Kim, Kyung H.

In: Journal of Structural Biology, Vol. 131, No. 1, 25.09.2000, p. 79-81.

Research output: Contribution to journalArticle

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