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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。
Low molecular weight heparin larger molecular heparin anticoagulant effect is small, the incidence of adverse reactions such as bleeding is low, with better clinical application prospects. In this paper, the cells of Flavobacterium heparinus were treated with anaerobic treatment, and the effects of temperature, pH value, aeration volume, amount of Tween 80 and lipolysis time on the growth and induction of enzyme production were studied. Factors, respectively, to determine the best conditions for bacterial culture and induction of enzyme production. The method of cell immobilization was also preliminarily studied. The immobilized cells could be reused after being embedded in sodium alginate, and the recovery rate of enzyme activity reached 36.1%. The immobilized cells were used to lyse the macromolecule heparin, and the molecular weight of about 5000 was observed by the final electrophoresis. It was further proved that the immobilized cells could produce low molecular weight heparin.