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• W1953670791 abstract "Some starch‐degrading enzymes accommodate carbohydrates at sites situated at a certain distance from the active site. In the crystal structure of barley α‐amylase 1, oligosaccharide is thus bound to the ‘sugar tongs’ site. This site on the non‐catalytic domain C in the C‐terminal part of the molecule contains a key residue, Tyr380, which has numerous contacts with the oligosaccharide. The mutant enzymes Y380A and Y380M failed to bind to β‐cyclodextrin‐Sepharose, a starch‐mimic resin used for α‐amylase affinity purification. The K d for β‐cyclodextrin binding to Y380A and Y380M was 1.4 m m compared to 0.20–0.25 m m for the wild‐type, S378P and S378T enzymes. The substitution in the S378P enzyme mimics Pro376 in the barley α‐amylase 2 isozyme, which in spite of its conserved Tyr378 did not bind oligosaccharide at the ‘sugar tongs’ in the structure. Crystal structures of both wild‐type and S378P enzymes, but not the Y380A enzyme, showed binding of the pseudotetrasaccharide acarbose at the ‘sugar tongs’ site. The ‘sugar tongs’ site also contributed importantly to the adsorption to starch granules, as K d = 0.47 mg·mL −1 for the wild‐type enzyme increased to 5.9 mg·mL −1 for Y380A, which moreover catalyzed the release of soluble oligosaccharides from starch granules with only 10% of the wild‐type activity. β‐cyclodextrin both inhibited binding to and suppressed activity on starch granules for wild‐type and S378P enzymes, but did not affect these properties of Y380A, reflecting the functional role of Tyr380. In addition, the Y380A enzyme hydrolyzed amylose with reduced multiple attack, emphasizing that the ‘sugar tongs’ participates in multivalent binding of polysaccharide substrates." @default.
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• W1953670791 date "2007-09-04" @default.
• W1953670791 modified "2023-10-18" @default.
• W1953670791 title "The ‘pair of sugar tongs’ site on the non-catalytic domain C of barley α-amylase participates in substrate binding and activity" @default.
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• W1953670791 doi "https://doi.org/10.1111/j.1742-4658.2007.06024.x" @default.
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