论文总字数：16472字

摘 要

自然界中最普遍的一类未被人们有效利用起来的碳水化合物纤维素。想要有效的利用它们，我们就需要得到纤维素酶，而里氏木霉在生产纤维素酶方面具有很多优点。但因为纤维素酶是一种诱导酶，碳源对所产酶的活性及酶组分有显著影响，所以本文探究了比较了里氏木霉对碳源、氮源、固定化载体等条件不同，所获得的纤维素酶活差异。进行了对里氏木霉产纤维素酶进行了液体摇瓶发酵实验，并用条件控制法对发酵培养基进行优化。采用了A滤纸酶活力（FPA）测定方法测定纤维素酶活力，DNS法测定还原糖浓度。实验结果表明：①.以乳糖为底物的游离和固定化发酵酶活差别不明显；②.无论是以乳糖或微晶纤维素为底物，加麸皮均有利于提高酶活；③.以微晶纤维素为底物的固定化发酵酶活远高于游离发酵；④.乳糖量高，发酵产的酶活相应的也较高；⑤.加酵母粉对发酵酶活有明显提高的影响；⑥.乳糖分批添加对发酵酶活有提高作用；⑦.微晶纤维素发酵产的酶活远比乳糖的发酵效果好；⑧.固定化载体对纤维素酶发酵影响较大，且影响表现为载体孔隙越大所得纤维素酶活力越高；⑨.碳酸钙的添加对酶活有提高作用。

关键词：里氏木霉；发酵条件；纤维素酶

Immobilized on the Richter scale on trichoderma viride cellulase production optimization

Abstract

The most common type of nature has not been organized carbohydrates cellulose. Want to use them effectively, we need to get the cellulose enzyme, and the magnitude of trichoderma viride has many advantages in terms of production of cellulase. But because of cellulase is an inducible enzyme, carbon source and enzyme activity of enzyme production by components have a significant impact, so this article explores the comparison of the magnitude of trichoderma viride on carbon source, nitrogen source, the condition such as immobilized carrier, cellulose enzyme activity gained by the difference. Was carried out on the magnitude of trichoderma viride produces cellulose enzyme liquid shaking flask fermentation experiments, and optimize the fermentation conditions of control laws. Adopted A filter paper enzyme (FPA) determination method to determine the cellulose enzyme activity, DNS method for the determination of reducing sugar concentration. The experimental results show that: (1). Lactose as the substrate of the free and immobilized fermentation enzyme activity difference is not obvious; (2). Whether to lactose or microcrystalline cellulose as the substrate, and bran to improve enzyme activity; (3). With microcrystalline cellulose as the substrate fermentation of immobilized enzyme activity was much higher than the free fermentation; (4). High lactose, fermentation to produce the enzyme activity of the corresponding also higher; 5. Add yeast powder has obviously improved the effect on fermentation enzyme activity; 6. Partial added lactose increase for fermentation, enzyme activity; All landowners. Microcrystalline cellulose fermentation to produce the enzyme activity than lactose fermentation effect is good; Today. Had a greater influence on the fermentation of cellulose enzyme immobilization carrier, and the greater the performance as the carrier pore the cellulose enzyme activity is higher; Pet-name ruby. Calcium carbonate are added to improve the enzyme activity.

Key words: magnitude of trichoderma viride, The fermentation conditions; cellulase

目录

摘 要 I

Abstract II

第一章 引言 1

1.1 前言 1

1.2 里氏木霉及其纤维素酶 1

1.2.1 纤维素酶 1

1.2.2 里氏木霉 2

1.3 固定化细胞产酶 2

1.4 产酶条件优化 3

1.5 本课题研究思路和意义 3

第二章 材料与方法 5

2.1 实验材料 5

2.1.1 实验菌种 5

2.1.2 实验仪器 5

2.1.3 实验试剂 6

2.2 实验方法 7

2.2.1 实验一：不同发酵培养基对纤维素酶活影响 7

2.2.2 实验二：摇瓶实验条件优化 9

2.2.3 分析检测方法 10

第三章 结果与讨论 13

3.1 标准曲线制作及回归方程 13

3.1.1 DNS测残糖的标准曲线制作 13

3.1.2 DNS测酶活的标准曲线制作 14

3.2 条件优化结果及分析 15

3.2.1固定化载体筛选 15

3.2.2添加碳酸钙对发酵的影响 16

3.2.3 游离和固定化发酵对比及麸皮对发酵的影响 16

3.2.4 乳糖添加方式及氮源（酵母粉）对发酵的影响 17

3.2.5 微晶纤维素对发酵的影响 17

第四章 结论与展望 19

4.1 结论 19

4.2 展望 19

参考文献 21

致谢 23

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