Open AccessCrystallization and preliminary X‐ray analysis of AAMS amidohydrolase, the final enzyme in degradation pathway I of pyridoxine
Author(s) -
Kobayashi Jun,
Yoshida Hiromi,
Chu Huy Nhat,
Yoshikane Yu,
Kamitori Shigehiro,
Yagi Toshiharu
Publication year - 2009
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309109026864
Subject(s) - amidohydrolase , crystallization , chemistry , enzyme , escherichia coli , monoclinic crystal system , stereochemistry , crystallography , biochemistry , crystal structure , organic chemistry , gene
α‐( 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 C 2, with unit‐cell parameters a = 393.2, b = 58.3, c = 98.9 Å, β = 103.4°, and diffraction data were collected to 2.7 Å 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.