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Publication Abstract from PubMed

beta-galactosidase (EC3.2.1.23) from Bacillus circulans ATCC 31382, designated BgaD, exhibits high transglycosylation activity to produce galacto-oligosaccharides. BgaD has been speculated to have a multiple domain architecture including a F5/8 type C domain or a discoidin (DS) domain in the C-terminal peptide region from amino acid sequence analysis. Here, we solved the first crystal structure of the C-terminal deletion mutant BgaD-D, consisting of sugar binding, Glyco_hydro, catalytic, and bacterial Ig-like domains, at 2.5 A. In the asymmetric unit, two molecules of BgaD-D were identified and the value of VM was estimated to be 5.0 A3 /Da. It had been speculated that the structure of BgaD-D consists of four domains. From the structural analysis, however, we clarified that BgaD-D consists of five domains. We identified a new domain structure comprised of beta-sheets in BgaD. The catalytic domain exhibits a TIM barrel structure with a small pocket suited for accommodating the disaccharides. Detailed structural information for the amino acid residues related to activity and substrate specificity was clarified in the catalytic domain. Furthermore, using the structural information, we successfully constructed some thermostable mutants via protein engineering method. This article is protected by copyright. All rights reserved.

Crystal structure of beta-galactosidase from Bacillus circulans ATCC 31382 (BgaD) and the construction of the thermophilic mutants.,Ishikawa K, Kataoka M, Yanamoto T, Nakabayashi M, Watanabe M, Ishihara S, Yamaguchi S FEBS J. 2015 Apr 16. doi: 10.1111/febs.13298. PMID:25879162^[1]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.