Linhardt R J, Fitzgerald G L, Cooney C L, Langer R
Biochim Biophys Acta. 1982 Apr 3;702(2):197-203. doi: 10.1016/0167-4838(82)90503-9.
Heparinase (heparin lyase, EC 4.2.2.7) prepared from Flavobacterium heparinum was used to digest heparin. The products of digestion were examined with a viscosometric assay at various stages of the reaction to measure their average molecular weight. By comparison with computer simulations of various models, heparinase was shown to act in a random endolytic mode. The relative abundance of intermediates in heparin degradation catalyzed by heparinase immobilized on Sepharose 4B was measured by high pressure liquid chromatography (HPLC) at various time points. The results obtained using HPLC were consistent with a random endolytic mechanism. The heparin digestion products were separated and identified using gel permeation chromatography. The final distributions of heparin degradation products for free and immobilized heparinase were identical. Contaminating sulfatases and glycuronidases which could have subsequently acted on heparin degradation products were not found in significant amounts in the heparinase preparation studied.
从肝素黄杆菌制备的肝素酶（肝素裂解酶，EC 4.2.2.7）用于消化肝素。在反应的各个阶段通过粘度测定法检查消化产物，以测量其平均分子量。通过与各种模型的计算机模拟进行比较，表明肝素酶以随机内切模式起作用。在不同时间点通过高压液相色谱（HPLC）测量固定在琼脂糖4B上的肝素酶催化的肝素降解过程中中间体的相对丰度。使用HPLC获得的结果与随机内切机制一致。使用凝胶渗透色谱法分离和鉴定肝素消化产物。游离和固定化肝素酶的肝素降解产物的最终分布是相同的。在所研究的肝素酶制剂中未发现可能随后作用于肝素降解产物的污染性硫酸酯酶和葡萄糖醛酸酶。
