Crystallization and preliminary X-ray studies of wild type and catalytic-site mutant alpha-amylase from Bacillus subtilis.
Recombinant alpha-amylase (EC3.2.1.1) from Bacillus subtilis has been crystallized by the hanging drop vapor diffusion method using polyethylene glycol as precipitant. Crystals of wild-type protein diffract to at least 2.2 A resolution, and belong to the space group P2(1)2(1)2(1) with a = 72.2 A, b = 74.9 A, c = 116.1 A with probably one molecule in the asymmetric unit. A catalytic site mutant created by site-directed mutagenesis has also been grown as isomorphous crystals with a = 72.6 A, b = 74.4 A, c = 116.7 A. Structural studies of both wild-type and mutant proteins will provide a basis for understanding the catalytic mechanism of alpha-amylase.
Mizuno
H
H
Department of Molecular Biology, National Institute of Agrobiological Resources, Ibaraki, Japan.
Morimoto
Y
Y
Tsukihara
T
T
Matsumoto
T
T
Takase
K
K
D001412:Bacillus subtilis / Q000201:enzymology*; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000648:ultrastructure; D001665:Binding Sites; D018360:Crystallography, X-Ray; D016297:Mutagenesis, Site-Directed; D011994:Recombinant Proteins; D013329:Structure-Activity Relationship; D000516:alpha-Amylases / Q000737:chemistry / Q000235:genetics / Q000648:ultrastructure*
D001426:Bacterial Proteins; D011994:Recombinant Proteins; D000516:alpha-Amylases