Summary
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1.
The fact that β-cyclodextrin inhibits the hydrolysis of amylase suggests that β-cyclodextrin binds to the active center of pancreatic α-amylase EC 3.2.1.1.
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2.
As shown by preparative ultracentrifugation, amylase binds specifically three moles of β-cyclodextrin per mole enzyme and the binding can be characterized by a single dissociation constant.
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3.
The dissociation constant of amylase-β-cyclodextrin complex determined by kinetic methods is KI = 200 µm, which agrees well with the value KD = 140 µm, determined by preparative ultracentrifugation, and with the value KS = 220 µm arrived at by spectrophotometric titration.
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4.
Solvent perturbation studies indicate that out of the three bound β-cyclodextrin molecules only one interacts with a tryptophyl side chain of amylase.
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Móra, S., Simon, I. & Elödi, P. Studies on the active center of pancreatic amylase. Mol Cell Biochem 4, 205–209 (1974). https://doi.org/10.1007/BF01731482
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DOI: https://doi.org/10.1007/BF01731482