Expression, crystallization and preliminary X-ray crystallographic study of ethanolamine ammonia-lyase from Escherichia coli.
Ethanolamine ammonia-lyase (EAL) catalyzes the adenosylcobalamin-dependent conversion of ethanolamine to acetaldehyde and ammonia. The wild-type enzyme shows a very low solubility. N-terminal truncation of the Escherichia coli EAL beta-subunit dramatically increases the solubility of the enzyme without altering its catalytic properties. Two deletion mutants of the enzyme [EAL(betaDelta4-30) and EAL(betaDelta4-43)] have been overexpressed, purified and crystallized using the sitting-drop vapour-diffusion method. Crystals of EAL(betaDelta4-30) and EAL(betaDelta4-43) diffracted to approximately 8.0 and 2.1 A resolution, respectively.
Department of Life Science, Graduate School of Science, University of Hyogo, 3-2-1 Koto, Kamigori-cho, Ako-gun, Hyogo 678-1297, Japan. shibach@sci.u-hyogo.ac.jp
Naoki
N
Shibata
Hiroko
H
Tamagaki
Shungo
S
Ohtsuki
Naoki
N
Hieda
Keita
K
Akita
Hirofumi
H
Komori
Yasuhito
Y
Shomura
Shin-ichi
S
Terawaki
Tetsuo
T
Toraya
Noritake
N
Yasuoka
Yoshiki
Y
Higuchi
D003460:Crystallization; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000201:enzymology*; D004981:Ethanolamine Ammonia-Lyase / Q000737:chemistry* / Q000235:genetics; D015870:Gene Expression; D009154:Mutation
D004981:Ethanolamine Ammonia-Lyase