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Publication Abstract from PubMed

The alpha-glucosidase from sugar beet (SBG) is an exo-type glycosidase. The enzyme has a pocket-shaped active site, but efficiently hydrolyzes longer maltooligosaccharides and soluble starch due to lower Km and higher kcat/Km for such substrates. To obtain structural insights into the mechanism governing its unique substrate specificity, a series of acarviosyl-maltooligosaccharides was employed for steady-state kinetic and structural analyses. The acarviosyl-maltooligosaccharides have a longer maltooligosaccharide moiety compared to the maltose moiety of acarbose, which is known to be the transition-state analog of alpha-glycosidases. The clear correlation obtained between logKi of the acarviosyl-maltooligosaccharides and log(Km/kcat) for hydrolysis of maltooligosaccharides suggests that the acarviosyl-maltooligosaccharides are transition state mimics. The crystal structure of the enzyme bound with acarviosyl-maltohexaose reveals that substrate binding at a distance from the active site is maintained largely by van der Waals interactions, with the four glucose residues at the reducing terminus of acarviosyl-maltohexaose retaining a left-handed single-helical conformation, as also observed in cycloamyloses and single-helical V-amyloses. The kinetic behavior and structural features suggest that the subsite structure suitable for the stable conformation of amylose lowers the Km for long-chain substrates, which in turn is responsible for higher specificity of the longer substrates.

Structural advantage of sugar beet alpha-glucosidase to stabilize the Michaelis complex with long-chain substrate.,Tagami T, Yamashita K, Okuyama M, Mori H, Yao M, Kimura A J Biol Chem. 2014 Dec 1. pii: jbc.M114.606939. PMID:25451917^[1]

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