Crystallization and preliminary X-ray diffraction experiments of arylmalonate decarboxylase from Alcaligenes bronchisepticus.
Arylmalonate decarboxylase catalyses the enantioselective decarboxylation of alpha-aryl-alpha-methylmalonates to produce optically pure alpha-arylpropionates. The enzyme was crystallized with ammonium sulfate under alkaline pH conditions with the aim of understanding the mechanism of the enantioselective reaction. X-ray diffraction data collected to a resolution of 3.0 A at cryogenic temperature showed that the crystals belonged to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 83.13, b = 99.62, c = 139.64 A. This suggested that the asymmetric unit would contain between four and six molecules. Small-angle X-ray scattering revealed that the enzyme exists as a monomer in solution. Thus, the assembly of molecules in the asymmetric unit was likely to have been induced during the crystallization process.
Nakasako
Masayoshi
M
Department of Physics, Faculty of Science and Technology, Keio University, 3-14-1 Hiyoshi, Kohoku-ku, Kanagawa 223-8522, Japan. nakasako@phys.keio.ac.jp
Obata
Rika
R
Okubo
Ryosuke
R
Nakayama
Shyuichi
S
Miyamoto
Kenji
K
Ohta
Hiromichi
H
D000421:Alcaligenes / Q000201:enzymology*; D001426:Bacterial Proteins / Q000032:analysis / Q000737:chemistry* / Q000302:isolation & purification; D002262:Carboxy-Lyases / Q000032:analysis / Q000737:chemistry* / Q000302:isolation & purification; D003460:Crystallization; D005615:Freezing; D014961:X-Ray Diffraction
D001426:Bacterial Proteins; D002262:Carboxy-Lyases; C071852:malonate decarboxylase