title: Crystallization and preliminary X-ray analysis of AAMS amidohydrolase, the final enzyme in degradation pathway I of pyridoxine.
abstract: alpha-(N-Acetylaminomethylene)succinic acid (AAMS) amidohydrolase from Mesorhizobium loti MAFF303099, which is involved in a degradation pathway of vitamin B(6) and catalyzes the degradation of AAMS to acetic acid, ammonia, carbon dioxide and succinic semialdehyde, has been overexpressed in Escherichia coli. To elucidate the reaction mechanism based on the tertiary structure, the recombinant enzyme was purified and crystallized by the sitting-drop vapour-diffusion method using PEG 8000 as precipitant. A crystal of the enzyme belonged to the monoclinic space group C2, with unit-cell parameters a = 393.2, b = 58.3, c = 98.9 A, beta = 103.4 degrees , and diffraction data were collected to 2.7 A resolution. The V(M) value and calculation of the self-rotation function suggested that three dimers with a threefold symmetry were possibly present in the asymmetric unit.
authors: Faculty of Agriculture and Agricultural Science Program, Graduate School of Integral Arts and Science, Kochi University, Nankoku, Kochi 783-8502, Japan.
authors: Jun
authors: J
authors: Kobayashi
authors: Hiromi
authors: H
authors: Yoshida
authors: Huy Nhat
authors: HN
authors: Chu
authors: Yu
authors: Y
authors: Yoshikane
authors: Shigehiro
authors: S
authors: Kamitori
authors: Toshiharu
authors: T
authors: Yagi
mesh_terms: D000581:Amidohydrolases / Q000737:chemistry* / Q000378:metabolism; D003460:Crystallization; D018360:Crystallography, X-Ray; D004591:Electrophoresis, Polyacrylamide Gel; D006868:Hydrolysis; D011487:Protein Conformation; D011736:Pyridoxine / Q000378:metabolism*
chemical_list: D000581:Amidohydrolases; D011736:Pyridoxine