Clinch Keith, Evans Gary B, Furneaux Richard H, Rendle Phillip M, Rhodes Phillippa L, Roberton Anthony M, Rosendale Douglas I, Tyler Peter C, Wright Damian P
Industrial Research Ltd., Gracefield Road, PO Box 31-310, Lower Hutt, New Zealand.
Carbohydr Res. 2002 Jun 12;337(12):1095-111. doi: 10.1016/s0008-6215(02)00104-0.
A chromogenic substrate, 4-nitrophenyl 2-acetamido-2-deoxy-beta-D-glucopyranoside 6-sodium sulfate was synthesized and used in combination with beta-N-acetylhexosaminidase for detection of the sulfatase, MdsA, by release of 4-nitrophenol. MdsA was originally isolated from the bacterium Prevotella strain RS2 and is believed to be involved in desulfation of sulfomucins, major components of the mucus barrier protecting the human colon surface. The exo nature of the MdsA sulfatase was indicated by its inability to de-esterify the disaccharide 4-nitrophenyl beta-D-galactopyranosyl-(1-->4)-2-acetamido-2-deoxy-beta-D-glucopyranoside 6-sodium sulfate. This latter compound was prepared from monosaccharide precursors by two different methods, the shorter requiring just six steps from 4-nitrophenyl 2-acetamido-2-deoxy-beta-D-glucopyranoside and giving an overall yield of 26.4%. The syntheses of 4-nitrophenyl beta-D-galactopyranoside 3-triethylammonium sulfate and 6-triethylammonium sulfate and their use in combination with beta-galactosidase as chromogenic substrates for detecting Bacteroides fragilis sulfatases with different specificities was also demonstrated.
合成了一种显色底物4-硝基苯基2-乙酰氨基-2-脱氧-β-D-吡喃葡萄糖苷6-硫酸钠，并将其与β-N-乙酰己糖胺酶联合使用，通过4-硝基苯酚的释放来检测硫酸酯酶MdsA。MdsA最初是从普雷沃氏菌RS2菌株中分离出来的，据信它参与了硫黏蛋白的脱硫过程，硫黏蛋白是保护人类结肠表面的黏液屏障的主要成分。MdsA硫酸酯酶的外切性质表现在它无法使二糖4-硝基苯基β-D-吡喃半乳糖基-(1→4)-2-乙酰氨基-2-脱氧-β-D-吡喃葡萄糖苷6-硫酸钠脱酯。后一种化合物由单糖前体通过两种不同方法制备，较短的方法从4-硝基苯基2-乙酰氨基-2-脱氧-β-D-吡喃葡萄糖苷开始只需六步，总产率为26.4%。还展示了4-硝基苯基β-D-吡喃半乳糖苷3-三乙铵硫酸盐和6-三乙铵硫酸盐的合成及其与β-半乳糖苷酶联合用作显色底物以检测具有不同特异性的脆弱拟杆菌硫酸酯酶的用途。
