几丁质酶的分离纯化及其降解性质的性质研究

文档编号:SW162  文档字数:11543,页数:27

 摘要：几丁质是广泛存在于自然界的一种含氮多糖类生物高分子，其降解产物壳寡糖具有许多优异的性能，已被应用于生产生活等各方面。几丁质酶是一种降解几丁质的专一性降解酶，降解产物为小分子片段的壳寡糖。本文主要讲述从嗜热芽孢杆菌HD1发酵产生的粗酶液出发，通过(NH4)2SO4沉淀、DEAE阴离子交换树脂层析和Sephacryl S-400凝胶柱层析，得到纯化后的几丁质酶，该酶的相对分子质量为80.8KD，具有较好的pH稳定性和较高的酶活力。经过薄层层析对酶解产物的分析表明，此酶对底物几丁质的切割方式为内切且降解产物具有良好的水溶解性。
 关键词：几丁质酶；分离纯化；酶解
 
 Abstract: Chitin is widely distributed in nature, a nitrogen-containing polysaccharide biopolymer.Chitosan degradation products have many excellent properties, and have been used in production and other aspects of life.Chitinase is an enzyme which catalyze the glycosidic band of chitin specificitily,and its product is small fragments of chitin.This thesis describes the processes of extracting chitinase produced by HD1 bacteria. The purified enzyme was purified by precipitation,DEAE-Sephadex ion-exchang chromatography and Sephacryl S-400 gel filtration.Its molecular mass was estimated to be about 80.8KD by SDS-PAGE. The hydrolysis product assayed by TLC suggested that the chitinase catalyzed an endo-type cleavage reaction,and the hydrolysis product was water solubility.

Keywords： Chitinase; Purification; Hydrolysis 
 

目录
中文摘要 I
英文摘要 II
目录 III
1.绪论 1
 1.1 壳寡糖在医药卫生方面的应用 2
 1.2 壳寡糖在农业中的应用 3
 1.3 壳寡糖在食品工业的应用 3
 1.4 壳寡糖在化妆品方面的应用 3
 1.5 壳寡糖的制备方法 3
2.实验部分 7
 2.1实验材料和仪器设备 7
 2.2 几丁质粗酶的制取 9
  2.3 几丁质粗酶的纯化 10
 2.4 薄层层析分析酶解产物 11
3.结果与讨论 13
 3.1 浓缩酶液最佳盐析浓度 13
 3.2 DEAE-Sepharose色谱图 14
 3.3 Sephacryl S-400色谱图 15
 3.4 酶纯度鉴定和分子量测定 16
 3.5 薄层层析分析酶解产物结果 17
4.总结与展望 22
致谢 23
参考文献 24

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