NDLI: Overexpression, purification, crystallization and preliminary structural studies of p-coumaric acid decarboxylase from Lactobacillus plantarum.

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Overexpression, purification, crystallization and preliminary structural studies of p-coumaric acid decarboxylase from Lactobacillus plantarum.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Rodríguez, Héctor de las Rivas, Blanca Muñoz, Rosario Mancheño, José M.
Description	Country affiliation: Spain Author Affiliation: Rodríguez H ( Instituto de Fermentaciones Industriales, CSIC, Juan de la Cierva 3, 28006 Madrid, Spain.)
Abstract	The substrate-inducible p-coumaric acid decarboxylase (PDC) from Lactobacillus plantarum has been overexpressed in Escherichia coli, purified and confirmed to possess decarboxylase activity. The recombinant His(6)-tagged enzyme was crystallized using the hanging-drop vapour-diffusion method from a solution containing 20%(w/v) PEG 4000, 12%(w/v) 2-propanol, 0.2 M sodium acetate, 0.1 M Tris-HCl pH 8.0 with 0.1 M barium chloride as an additive. Diffraction data were collected in-house to 2.04 A resolution. Crystals belonged to the tetragonal space group P4(3), with unit-cell parameters a = b = 43.15, c = 231.86 A. The estimated Matthews coefficient was 2.36 A(3) Da(-1), corresponding to 48% solvent content, which is consistent with the presence of two protein molecules in the asymmetric unit. The structure of PDC has been determined by the molecular-replacement method. Currently, the structure of PDC complexed with substrate analogues is in progress, with the aim of elucidating the structural basis of the catalytic mechanism.
File Format	HTM / HTML
ISSN	2053230X
e-ISSN	17443091
Journal	Acta Crystallographica Section F Structural Biology and Crystallization Communications
Issue Number	Pt 4
Volume Number	63
Language	English
Publisher	Wiley
Publisher Date	2007-04-01
Publisher Place	Great Britain (UK)
Access Restriction	Open
Subject Keyword	Discipline Crystallography Discipline Biophysics Discipline Molecular Biology Discipline Biochemistry Carboxy-lyases Chemistry Lactobacillus Plantarum Enzymology Amino Acid Sequence Genetics Isolation & Purification Chromatography, Affinity Crystallization Crystallography, X-ray Dna Primers Electrophoresis, Polyacrylamide Gel Escherichia Coli Molecular Sequence Data Protein Conformation Recombinant Proteins Research Support, Non-u.s. Gov't
Content Type	Text
Resource Type	Article
Subject	Genetics Structural Biology Medicine Biochemistry Biophysics Condensed Matter Physics

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