使用β-麦芽糖苷（对硝基苯基-β-D-麦芽糖苷、2-氯-4-硝基苯基-β-D-麦芽糖苷和4-甲基伞形酮基-β-D-麦芽糖苷）作为底物测定人肾和尿液中的中性α-葡萄糖苷酶。

Use of beta-maltosides (p-nitrophenyl-beta-D-maltoside, 2-chloro-4- nitrophenyl-beta-D-maltoside and 4-methylumbelliferyl-beta-D-maltoside) as substrates for the assay of neutral alpha-glucosidase from human kidney and urine.

作者信息

Lukomskaya I S, Voznyi Y V, Lanskaya I M, Podkidisheva E I

机构信息

Institute of Biomedical Chemistry of RAMS, Moscow, Russian Federation.

出版信息

Clin Chim Acta. 1996 Jan 31;244(2):145-54. doi: 10.1016/0009-8981(95)06189-4.

DOI:10.1016/0009-8981(95)06189-4

PMID:8714432

Abstract

The method of assay of neutral alpha-glucosidase from human kidney and urine using beta-maltosides (p-nitrophenyl-beta-D-maltoside [NP-beta-D-maltoside], 2-chloro-4-nitrophenyl-beta-D-maltoside]) [CNP-beta-D-maltoside] and 4-methylumbelliferyl-beta-D-maltosides ([MU-beta-D-maltoside]) as substrates and beta-glucosidase as an auxiliary enzyme is proposed. All three beta-maltosides are suitable substrates for the determination of neutral alpha-glucosidase activity but MU-beta-D-maltoside is the most sensitive due to its methylumbelliferyl moiety. The method is simple, convenient and 10-fold more sensitive than the commonly used alpha-glucosidase assay procedure with the corresponding synthetic alpha-glucosides, p-nitrophenyl- alpha-D-glucoside (NP-alpha-D-glucoside) and 4-methylumbelliferyl-alpha-D-glucoside (MU-alpha-D-glucoside). A modification of the method, with p-nitrophenyl-beta-D-maltoside as substrate, was applied to the semiautomatic assay of urinary alpha-glucosidase in 96-well microtitre plates.

摘要

提出了一种以β-麦芽糖苷（对硝基苯基-β-D-麦芽糖苷[NP-β-D-麦芽糖苷]、2-氯-4-硝基苯基-β-D-麦芽糖苷[CNP-β-D-麦芽糖苷]）和4-甲基伞形酮基-β-D-麦芽糖苷[（MU-β-D-麦芽糖苷]）为底物，β-葡萄糖苷酶为辅助酶，用于检测人肾和尿液中中性α-葡萄糖苷酶的方法。所有这三种β-麦芽糖苷都是测定中性α-葡萄糖苷酶活性的合适底物，但由于其甲基伞形酮基部分，MU-β-D-麦芽糖苷最为灵敏。该方法简单、方便，比常用的以相应的合成α-葡萄糖苷对硝基苯基-α-D-葡萄糖苷（NP-α-D-葡萄糖苷）和4-甲基伞形酮基-α-D-葡萄糖苷（MU-α-D-葡萄糖苷）检测α-葡萄糖苷酶的方法灵敏10倍。以对硝基苯基-β-D-麦芽糖苷为底物的该方法的一种改进形式，被应用于在96孔微量滴定板中半自动检测尿液α-葡萄糖苷酶。