【作者】 张鑫欣；

【导师】 董明盛；

【作者基本信息】 南京农业大学 ， 食品科学， 2006， 硕士

【摘要】 纳豆激酶（Nattokinase）是由纳豆菌在发酵大豆过程中产生的一种具有纤溶活性的丝氨酸蛋白酶。国内外研究表明，在纤维蛋白平板试验、动物血栓模型试验和临床试验中，纳豆激酶都表现了明显的溶栓作用。其通过食品发酵而来，在胃肠环境中不会失活，可通过消化道吸收，不仅溶栓效率高、疗效时间长，且安全可靠。将纳豆激酶制品开发成口服型溶栓药物及保健食品的前景广阔。 液芯海藻酸钙微胶囊是起源于海藻酸钙凝胶珠的一种固定化体系，它不仅具有海藻酸钙凝胶珠的来源广泛，价格低廉，固定化条件温和，操作简单，生物相容性好，基质和产物扩散阻力小等众多优点，而且还能克服凝胶珠的营养物质和氧气不易传递到内部，细胞只能生长在胶珠表面和凝胶孔中等缺点，因而是一种有广阔发展前景的新型微胶囊体系。 多孔淀粉又称为微孔淀粉，孔径在1μm左右，能同时吸附多种物质。在微胶囊的液芯中加入了多孔淀粉则进一步增强了微胶囊的包埋能力，产生良好的固定化作用。 本文首次研究了将生物微胶囊和多孔淀粉同时应用于纳豆菌及纳豆激酶的固定化，确定其最优制囊条件，并研究了生物微胶囊各制囊成分及多孔淀粉与纳豆菌的生物相容性，采用响应曲面法（Response Surface Methodology，RSM）优化了产酶条件，并通过发酵罐放大试验进一步确定了固定化纳豆菌的最优发酵条件，为生物微胶囊／多孔淀粉固定化纳豆菌在纳豆激酶工业发酵中的应用打下基础。主要结果如下： 1．研究了生物微胶囊／多孔淀粉固定纳豆菌的最适制囊条件，结论为：海藻酸钠含量为0.8％、CaCl₂含量为1.5％时产酶效果最好，多孔淀粉含量为4.0％，黄原胶含量为0.25％时产酶效果与制囊条件的可行性达到最佳。并确定了微胶囊囊材与纳豆菌之间具有良好的生物相容性。 2．采用Plackett-Burman设计（Plackett-Burman Design，PB）与中心组合设计（Central Composite Design，CCD）法，对纳豆菌DU115的发酵培养基，对影响纳豆菌发酵液纳豆激酶活性的9个相关因素进行了研究和探讨。结果表明：影响DU115发酵液纳豆激酶活性的培养基组分的关键因子为发豆粕粉、葡萄糖、MgSO₄；在其它更多还原

【Abstract】 Nattokinase, a kind of serine enzyme produced by Bacillus subtilis natto during fermentation, has fibrinolytic activity. Studies showed that nattokinase could obviously dissolve fibrin and blood clot, enhance fibrinolytic system’s function of animal and human thrombus model. Nattokinase from fermented food with high efficiency of fibrinolytic was stable in enteron and easy to absorb by intestine. Therefore development of nattokinase for healthy food and oral thrombolysis medicament had a bright prospect.Intra-hollow Ca-alginate microcapsule comes of the Ca-alginate bead. It not only owns the advantages of broad source, inexpensive cost, gentle immobilization condition, easy operation, good biocompatibility, and less resistance to the substrates and the oxygen, but also can overcome the disadvantages of the difficulty that the substrates and oxygen can hardly transfer into the bead core and the limited space that the cells can only grow on the bead surface and in the gel holes. Therefore, the intra-hollow Ca-alginate capsule is a promising immobilization carrier.Microporous starch can absorb materials in its micropore. Put microporous starch into intra-hollow Ca-alginate capsule can augment the content ability of microcapsule.Biomicrocapsule and microporous starch used to immobilize Bacillus subtilis natto and Nattokinase in the same time was studied for the first time in this thesis. Response Surface Methodology was used to optimize the fermentation condition of immobilized Nattokinase. Also the enzyme activity of Nattokinase in fermentation tank and three batches continuous culture fermentation was studied in this thesis. The main results were as follows:1. The preparation of Biomicrocapsule and microporous starch immobilizing Bacillus subtilis natto was studied and the result was: when the concentration Na -alginate 0.8%, CaCl₂ 1.5%, Microporous starch 4.0%, Xanthan 0.25%, the preparation was feasible and更多还原