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Título

Overexpression, purification, crystallization and preliminary structural studies of catabolic ornithine transcarbamylase from Lactobacillus hilgardii

AutorRivas, Blanca de las ; Rodríguez, Héctor; Angulo, Iván ; Muñoz, Rosario ; Mancheño, Jose M.
Palabras clavecatabolic ornithine transcarbamylase
Lactobacillus hilgardii
arginine degradation
Fecha de publicación2007
EditorJohn Wiley & Sons
CitaciónActa Crystallographica Section F : Structural Biology and Crystallization Communications 63(7): 563-567 (2007)
ResumenThe catabolic ornithine transcarbamylase (cOTC; EC 2.1.3.3) from the lactic acid bacteria Lactobacillus hilgardii is a key protein involved in the degradation of arginine during malolactic fermentation. cOTC containing an N-terminal His6 tag has been overexpressed in Escherichia coli, purified and crystallized under two different experimental conditions using the hanging-drop vapour-diffusion method. Crystals obtained from a solution containing 8%(w/v) PEG 4000, 75 mM sodium acetate pH 4.6 belong to the trigonal space group P321 and have unit-cell parameters a = b = 157.04, c = 79.28 Å. Conversely, crystals grown in 20%(v/v) 2-methyl-2,4-pentanediol, 7.5%(w/v) PEG 4000, 100 mM HEPES pH 7.8 belong to the monoclinic space group C2 and have unit-cell parameters a = 80.06, b = 148.90, c = 91.67 Å, [beta] = 100.25°. Diffraction data were collected in-house to 3.00 and 2.91 Å resolution for trigonal and monoclinic crystals, respectively. The estimated Matthews coefficient for the crystal forms were 2.36 and 2.24 Å3 Da-1, respectively, corresponding to 48% and 45% solvent content. In both cases, the results are consistent with the presence of three protein subunits in the asymmetric unit. The structure of cOTC has been determined by the molecular-replacement method using the atomic coordinates of cOTC from Pseudomonas aeruginosa (PDB code 1dxh) as the search model
Versión del editorhttp://dx.doi.org/10.1107/S1744309107025195
URIhttp://hdl.handle.net/10261/46370
DOI10.1107/S1744309107025195
ISSN1744-3091
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