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Crystallization and preliminary X-ray studies of wild type and catalytic-site mutant alpha-amylase from Bacillus
H Mizuno1, Y Morimoto, T Tsukihara
1Department of Molecular Biology, National Institute of Agrobiological Resources, Ibaraki, Japan.
Abstract:
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.