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摘要：本实验在纤维素降解菌能把纤维素降解为葡萄糖的特性下，用DNS法测定含纤维素降解菌的培养基中还原糖的含量来比较不同纤维素降解菌降解能力的大小。首先通过筛选出较为优势的纤维素降解菌进行纯培养，然后在以纤维素为唯一碳源的培养基上对各种菌种进行摇瓶培养并测定不同时期不同菌种的还原糖含量，最后通过还原糖含量计算出酶活来进行比较确定酶活大小。结果表明，NF 16-3和 NF 7-2的酶活力较高，NF 6-1和 NF 5-2的酶活力较低。通过酶活测定实验，对纤维素降解菌某些种类有了一定的了解，为纤维素酶进一步用于工业化生产奠定了基础。59239
毕业论文关键词：DNS法，纤维素降解菌，纤维素酶，酶活测定及比较
Abstract: The experiment is based on characteristics that the cellulose degradation bacteria can degradate the cellulose into glucose. Using the method of DNS determination to measure the Sugar content of Containing cellulose degrading bacteria on the medium to compare the size of the different ability of degradation of cellulose degradation bacteria. First screen the cellulose degradation bacteria with more advantage and do pure culture, and then do shake flask culture of various strains and determine the reducing sugar contenting in different strains of different period on the medium which take cellulose as the sole carbon source. At last, compare and determine the size of the enzyme afer calculating the contenting reducing sugar.The results showed that enzyme NF 16-3 and NF 7-2 is more active, while enzyme NF 6-1 and NF 5-2 is less active. Through experiment for the Determination of enzyme activity , a better understanding of certain kinds of cellulose degradation bacteria is gained, which lays a
groundwork for future industrialized production of cellulase.
Keywords: DNS method, cellulose degrading bacteria, cellulose enzyme, enzyme activity determination and comparison
目 录
1 前言 4
2 实验材料与方法 5
2.1 菌种 5
2.2 培养基 5
2.2.1 纤维素培养基 5
2.3主要仪器和试剂 5
2.3.1仪器： 5
2.4实验方法 5
2.4.1培养基的处理 5
2.4.2缓冲液的制备 6
2.4.3菌种活化 6
2.4.4液体摇瓶培养 6
2.4.5粗酶液的制备 6
2.4.6酶活测定 6
3结果与分析 7
3.1葡萄糖标准曲线的配置 7
3.1.1葡糖糖标准液的配置 7
3.1.2葡萄糖标准曲线的绘制 8
3.2酶活测定 8
3.2.1不同时间对同种菌菌液酶活力的影响 8
3.2.2同一时间对不同菌种菌液酶活力的影响 11
3.2.3不同时间对不同菌种菌液酶活力的影响 13
3.2.4菌种菌液酶活与菌种菌体酶活比较 14
3.3不同时期菌液情况比较 16
结 论 18
参考文献 19