Alpha-glucosidase inhibition assay in an enzyme-immobilized amino-microplate.
Alpha-glucosidase (AGH) from the small intestine of rat was immobilized onto a glutaraldehyde (GA) activated NH(2)-96 well microplate to establish a convenient and rapid AGH inhibition assay system. After AGH immobilization, remaining GA groups were blocked by beta-alanine to induce a negative charge on the surface of the well. The AGH-plate showed an enzyme activity of 444 nU/well under an assayed condition at 37 degrees C for 2 h using 0.3 mM 4-methylumbelliferyl-alpha-D-glucopyranoside as a fluorogenic substrate. Inhibitory powers of voglibose and acarbose as therapeutic AGH inhibitors were successfully evaluated to have IC(50) values of 13 and 114 nM, respectively.
Matsui
Toshiro
T
Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University, Higashi, Fukuoka 812-8581, Japan. tmatsui@agr.kyushu-u.ac.jp
Shimada
Mayu
M
Saito
Nozomi
N
Matsumoto
Kiyoshi
K
D020909:Acarbose / Q000494:pharmacology; D000818:Animals; D055162:Biocatalysis; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000037:antagonists & inhibitors* / Q000737:chemistry*; D005976:Glutaral / Q000737:chemistry*; D065089:Glycoside Hydrolase Inhibitors; D020128:Inhibitory Concentration 50; D007294:Inositol / Q000031:analogs & derivatives / Q000494:pharmacology; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D013997:Time Factors; D000520:alpha-Glucosidases / Q000737:chemistry* / Q000378:metabolism; D015091:beta-Alanine / Q000494:pharmacology
D004791:Enzyme Inhibitors; D004800:Enzymes, Immobilized; D065089:Glycoside Hydrolase Inhibitors; D015091:beta-Alanine; D007294:Inositol; D000520:alpha-Glucosidases; C102817:voglibose; D005976:Glutaral; D020909:Acarbose