肝素黄杆菌的固定化及其在低分子肝素制备中的探究

低分子肝素较大分子肝素抗凝血作用小,出血等不良反应发生率低,具有更好的临床应用前景。论文对肝素黄杆菌细胞进行缺壁处理,并考察了温度、pH值、通气量、吐温80用量及溶脂时间等因素对菌体生长和诱导产酶的影响,使用正交实验优化了各个因素,分别确定了菌体培养以及诱导产酶的最佳条件。还初步研究了细胞固定化方法,使用海藻酸钠包埋,得到可重复利用的固定化细胞,其酶活性回收率可达到36.1%。利用固定化细胞裂解大分子肝素,最终电泳观测到相对分子质量为5000左右的目标条带,进一步证明了利用固定化细胞生产低分子质量肝素具有可行性。

The Immobilization of Flavobacterium Heparinum Cells and the Inquiry of Its Application in the Preparation of Low Molecular Weight Heparin

Comparing with normal heparin,low molecular weight heparin(LMWH) has lots of advantages,such as smaller anticoagulant effect and lower rate of bleeding,hence it has a good prospect in clinical application.In this experiment,the cellular wall of Flavobacterium heparinum was partly dissolved to get cells with a higher permeability.Then the cells were immobilized and used to degrade high molecular weight heparin.The enzyme activity recovery of the immobilized cells reached a value of 36.1%.Finally apolyacrylamide gel electrophoresis(PAGE) was applied to detect product and the electrophoretic band of LMWH with an average molecular weight of 5000 was observed.In the whole process,orthogonal experimental design was used to optimize the condition and the optimum temperature,pH value,rotation speed,and the amount or incubation time of Tween 80 were all determined.