Rice K G, Linhardt R J
Division of Medicinal and Natural Products Chemistry, College of Pharmacy, University of Iowa, Iowa City 52242.
Carbohydr Res. 1989 Jul 15;190(2):219-33. doi: 10.1016/0008-6215(89)84127-8.
The rapid preparation of multimilligram quantities of five heparin-derived oligosaccharides (1-5) is described. These oligosaccharides are the final products obtained from the action of heparin lyase (heparinase, E.C. 4.2.2.7) at its primary sites in the heparin polymer. Five oligosaccharides comprise from 75-85 wt% of commercial porcine mucosal heparins and are recovered in good yield and high purity. Four of these five oligosaccharides were further acted upon at much lower rates by prolonged treatment with heparin lyase or heparan monosulfate lyase (heparitinase, E.C. 4.2.2.8), revealing the subspecificities of these enzymes. These oligosaccharides were used as defined substrates for heparin lyase and heparan monosulfate lyase and their kinetic constants were obtained. Potential applications for these oligosaccharides include their use as defined substrates for purification of heparin monosulfate lyases, and for establishing the catalytic purity of enzyme preparations.
本文描述了快速制备数毫克量的五种肝素衍生寡糖（1-5）的方法。这些寡糖是肝素裂解酶（肝素酶，E.C. 4.2.2.7）作用于肝素聚合物主要位点后得到的最终产物。这五种寡糖占商业猪黏膜肝素重量的75-85%，且能以高收率和高纯度回收。通过用肝素裂解酶或硫酸乙酰肝素单硫酸酯裂解酶（硫酸乙酰肝素酶，E.C. 4.2.2.8）长时间处理，这五种寡糖中的四种以低得多的速率进一步反应，揭示了这些酶的底物特异性。这些寡糖用作肝素裂解酶和硫酸乙酰肝素单硫酸酯裂解酶的特定底物，并获得了它们的动力学常数。这些寡糖的潜在应用包括用作纯化硫酸乙酰肝素单硫酸酯裂解酶的特定底物，以及用于确定酶制剂的催化纯度。
