摘 要：利用里氏木霉产纤维素酶，在工业生产中日趋重要。本论文研究的是里氏木霉产纤维素酶时种子培养的优化，实验分别以不同培养时间的种子，按照3%、5%、10%的接种量接种发酵瓶考察酶活变化；分别以添加不同C源、N源的种子培养及培养24h，按照3%、5%、10%的接种量接种发酵培养基考察酶活变化，以期通过最佳接种量和最佳种子获得较高的酶活。实验表明，种子最佳接种量为5%，培养18h的种子为最佳，获得较高酶活应在发酵时间≥132h后。实验为纤维素酶在解决环境污染问题和在饲料工业上的应用提供实验基础，也为实际生产中利用纤维素酶提高纤维素的利用率提供依据。57498

毕业论文关键词：里氏木霉Rut C-30，纤维素酶，酶活，种子优化

Abstract: Using the Trichoderma reesei as cellulase producer has become increasingly important in industrial cellulase production. This thesis is to optimize the inoculum of T. reesei as cellulase producer. The experiment uses inoculum with different incubation time (8h, 12h, 18h, 28h, 48h and 72h) and different amount (3%, 5% and 10% )of the fermentation medium or inoculum added different C sources and N sources into the inoculum medium with 3%, 5% and 10% of inoculation volume to observe the changes of cellulase activity over time. The purpose of this experiment is to achieve the best quantity inoculum to produce higher enzyme activity. The results show that the optimal inoculation volume is 5% with 18 hours incubation, and the cellulase activity starts to rapid increasing after 132 fermentation. This result could be used to solve the problem of environmental pollution and provide experimental basis for application in feed industry, as well as provide the basis of raising the utilization rate of cellulose by using cellulase enzyme in the actual production.

Keywords：Trichoderma reesei Rut C-30, cellulases, enzyme activity, inoculum optimization

目录

1  前言 3

1.1  研究背景 3

1.2  本文研究的目的和意义 4

2  材料与方法 4

2.1  实验材料 4

2.1.1  菌种 4

2.1.2  实验材料 4

2.1.3  仪器 5

2.2  实验方法 5

2.2.1  培养基 5

2.2.1.1  PDA培养基 5

2.2.1.2  玉米浆种子培养基 7

2.1.2.3  发酵基本培养基 7

2.2.2  培养方法 8

2.2.2.1  菌种制备 8

2.2.2.2  纤维素酶制备 8

2.3  试剂配制 8

2.3.1  柠檬酸缓冲液的配制 8

2.3.2  DNS试剂的配制 8

2.4  摇瓶发酵阶段实验设计 9

2.5  纤维素酶滤纸活力测定 9

2.5.1  原理 9

2.5.2  制备葡萄糖标准曲线 10

2.5.3  测滤纸酶活 10

3  结果和讨论 11

3.1  葡萄糖（G）标准曲线