用于测定2-乙酰氨基-2-脱氧-D-葡萄糖6-硫酸酯硫酸酯酶和（1→3）-N-乙酰-β-D-氨基葡萄糖苷酶的硫酸角质素底物的制备。

Preparation from keratin sulfate of substrates for the measurement of 2-acetamido-2-deoxy-D-glucose 6-sulfate sulfatase and (1 goes to 3)-N-acetyl-beta-D-glucosaminidase.

作者信息

Toma S, Di Ferrante D T, Tenni R, Di Ferrante N

出版信息

Carbohydr Res. 1981 Jan 15;88(1):93-105. doi: 10.1016/s0008-6215(00)84604-2.

Abstract

An extract of bacterial cells Pseudomonas sp. IFO-13309 grown on medium containing 0.1% bovine cornea keratan sulfate of low sulfate content degraded exhaustively bovine cornea keratan sulfate to give 2-acetamido-2-deoxy-beta-D-gluco-pyranosyl 6-sulfate-(1 goes to 3)-D-galactose, isolated by gel filtration on Sephadex G-25 and purified by preparative paper chromatography. This was reduced with sodium borotritide to give 2-acetamido-2-deoxy-beta-D-glucopyranosyl 6-sulfate-(1 goes to 3)-D-[1-3H]galactitol, purified by gel filtration on Sephadex G-15, which was an excellent substrate for the measurement of 2-acetamido-2-deoxy-D-glucose 6-sulfate sulfatase. The reduced, radioactive monosulfated disaccharide was desulfated with methanolic 70mM hydrogen chloride and purified by gel filtration on Sephadex G-15 to give O-(2-acetamido-2-deoxy-beta-D-glucopyranosyl)-(1 goes to 3)-D-[1-3H]galactitol, which allowed the measurement of (1 goes to 3)-N-acetyl-beta-D-glucosaminidase. This enzyme may participate in the normal degradation of keratan sulfate.

摘要

在含有0.1%低硫酸含量牛角膜硫酸角质素的培养基上生长的假单胞菌属IFO-13309细菌细胞提取物能将牛角膜硫酸角质素彻底降解，生成2-乙酰氨基-2-脱氧-β-D-吡喃葡萄糖基6-硫酸盐-(1→3)-D-半乳糖，通过在Sephadex G-25上进行凝胶过滤分离，并通过制备性纸色谱法纯化。用硼氢化三钠将其还原得到2-乙酰氨基-2-脱氧-β-D-吡喃葡萄糖基6-硫酸盐-(1→3)-D-[1-³H]半乳糖醇，通过在Sephadex G-15上进行凝胶过滤纯化，它是用于测定2-乙酰氨基-2-脱氧-D-葡萄糖6-硫酸盐硫酸酯酶的优良底物。还原后的放射性单硫酸化二糖用70mM甲醇盐酸脱硫，并通过在Sephadex G-15上进行凝胶过滤纯化得到O-(2-乙酰氨基-2-脱氧-β-D-吡喃葡萄糖基)-(1→3)-D-[1-³H]半乳糖醇，可用于测定(1→3)-N-乙酰-β-D-氨基葡萄糖苷酶。这种酶可能参与硫酸角质素的正常降解。