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Preparation of nonreducing end substituted p nitrophenyl alpha maltopentaoside fg 5p as a substrate for a coupled enzymatic assay for alpha amylase


, : Preparation of nonreducing end substituted p nitrophenyl alpha maltopentaoside fg 5p as a substrate for a coupled enzymatic assay for alpha amylase. Journal of Biochemistry (Tokyo) 97(4): 977-982

p-Nitrophenyl O-6-deoxy-6-[(2-pyridyl)amino]-.alpha.-D-glucopyranosyl-(1 .fwdarw. 4)-O-.alpha.-D-glucopyranosyl-(1 .fwdarw. 4)-O-.alpha.-D-glucopyranosyl-(1 .fwdarw. 4)-O-.alpha.-D-glucopyranosyl-(1 .fwdarw. 4)-.alpha.-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]-.alpha.-D-glucopyranosyl-(1 .fwdarw. 4)-O-.alpha.-D-glycopyranosyl-(1 .fwdarw. 4)-O-.alpha.-D-glucopyranosyl-(1 .fwdarw. 4)-O-.alpha.-D-glycopyranosyl-(1 .fwdarw. 4)-O-.alpha.-D-glucopyranosyl-(1 .fwdarw. 4)-D-glucose and p-nitrophenyl .alpha.-glucoside. The maltopentaose derivative is resistant to .alpha.-glucosidase and is suitable as a substrate for the .alpha.-amylase assay coupled with .alpha.-glucosidase in which the activity of .alpha.-amylase is determined by measuring the amount of p-nitrophenol liberated by .alpha.-glucosidase from p-nitrophenyl .alpha.-glucoside and p-nitrophenyl .alpha.-maltoside produced by the action of .alpha.-amylase. This .alpha.-amylase assay method was applied for determination of .alpha.-amylases in human serum.

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