Enzymatic synthesis of p-nitrophenyl 3(5)-O-beta-N-acetylglucosaminyl-alpha-maltopentaoside by lysozyme; a novel substrate for human amylase assay.
Transglycosylation from di-N-acetylchitobiose to the 3-position at the nonreducing end glucosyl group of p-nitrophenyl alpha-maltopentaoside was regioselectively induced through the use of hen egg-white lysozyme. The enzyme formed p-nitrophenyl 3(5)-O-beta-N-acetylglucosaminyl-alpha-maltopentaoside (5% of the enzyme-catalyzed net decrease of p-nitrophenyl alpha-maltopentaoside) from di-N-acetylchitobiose as a donor and p-nitrophenyl alpha-maltopentaoside as an acceptor. The rate of the transglycosylation depended on the concentration of substrate, the temperature and the pH. The hydrolytic actions of human pancreatic and salivary alpha-amylase on this derivative were examined. The maltopentaoside derivative was shown to be useful as a substrate for alpha-amylase assay through a coupled reaction involving alpha-D-glucosidase and glucoamylase.
Department of Applied Biochemistry, Faculty of Agriculture, Shizuoka University, Japan.
H
H
Matsui
H
H
Kawagishi
T
T
Usui
D000117:Acetylglucosamine / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000378:metabolism; D000681:Amylases / Q000032:analysis*; D002151:Calorimetry; D002240:Carbohydrate Sequence; D002851:Chromatography, High Pressure Liquid; D005944:Glucosamine / Q000031:analogs & derivatives*; D006801:Humans; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D009113:Muramidase / Q000378:metabolism*; D009844:Oligosaccharides / Q000138:chemical synthesis / Q000378:metabolism*; D013379:Substrate Specificity; D000520:alpha-Glucosidases / Q000378:metabolism*
D009844:Oligosaccharides; C065379:4-nitrophenyl-3(5)-O-N-acetylglucosaminyl-maltopentaoside; D000681:Amylases; D009113:Muramidase; D000520:alpha-Glucosidases; D005944:Glucosamine; D000117:Acetylglucosamine