Preparation of Non-Reducing-End Substituted p-Nitrophenyl α-Maltopentaoside (FG5P) as a Substrate for a Coupled Enzymatic Assay for α-Amylases

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Abstract
p-Nitrophenyl O-6-deoxy-6-[(2-pyridyl)amino]-α-D-glucopyranosyl-(1↑4)-O-α-D-glu-copyranosyl-(1↑4)-O-α-D-glucopyranosyl-(1↑4)-O-α-D-glucopyranosyl-(1—4)-α-D-glucopyranoside, FG5P, was prepared, taking advantage of the action of Bacillus macerans cyclodextrin glucanotransferase on a mixture of O-6-deoxy-6-[(2-pyridyl)-amino]-α-D-glucopyranosyl-(1↑4)-O-α-D-glucopyranosyl-(1↑4)-O-α-D-glucopyranosyl-(1↑4)-O-α-D-glucopyranosyl-(1↑4)-O-α-D-glucopyranosyl-(1↑4)-D-glucose and p-nitrophenyl α-glucoside. The maltopentaose derivative is resistant to α-glucosidase and is suitable as a substrate for the α-amylase assay coupled with α-glucosidase in which the activity of α-amylase is determined by measuring the amount of p-nitrophenyl liberated by α-glucosidase from p-nitrophenyl α-glucoside and p-nitrophenyl α-maltoside produced by the action of α-amylase. This α-amylase assay method was applied for determination of α-amylases in human serum.

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