木质素细菌降解酶的分离、筛选与优化本科毕业设计（论文）.docVIP

XX 本科毕业设计（论文） 题 目： 木质素细菌降解酶的 分离、筛选与优化 学 院： 专 业： 学 号： 学生姓名： 指导教师： 职 称： 二O一 年 月 日 摘 要 生物质原料作为地球上数量最大的可再生资源之一，逐渐受到人们的重视。目前，采用木质素细菌降解酶作为预处理微生物，正成为新的研究方向。本实验以南京紫金山土壤为原料，通过稀释平板法，固体培养基培养以及试管斜面保种等步骤，筛选得到纯种细菌。在筛选细菌过程中，挑选部分采用液体培养基培养，得到细菌降解酶，并测其漆酶和锰过氧化物酶的酶活力，结果发现酶活力都不高。最后通过正交试验优化菌28的产酶条件，发现在pH为3、Cu2+ 浓度为5mol/100mL、Mn2+浓度为1mg/100mL的条件下，细菌产漆酶量最大。 关键字： 木质素；稀释分离；酶活；优化 Abstract As one of the largest number of renewable resources on the earth, biological raw material has been paid more and more attention. At present, biological pretreatment used by bacteria lignin degrading enzymes is becoming a new kind of research interest. In this paper, raw material was obtained the soil in the Nanjing Zijin Mountain, and then finally lignin degradation bacterium were isolated. in test tubes which were preserved in refrigerator with suitable centigrade by means like dilution，screening and solid nutrient medium. During the process in screening,we selected some bacteria to be fostered in liquid nutrient medium and measured their laccase enzymes activity and manganese peroxidase activity.The result indicated that both laccase enzymes activity and manganese peroxidase activity were not high .At last, we tried to optimize the requirement that the bacteria No.28 need to produce enzymes by orthogonal test,and discovered that the bacteria could produce the largest quantity of Lac in the condition where the pH was 3,and the consistence of Cu2+ was 5mol/100mL,and the consistence of Mn2+ was 1mg/100mL. Key words: lignin，isolation，enzymes activity，optimization 目 录 第一章 绪论 1 1.1 本课题的目的、意义 1 1.2 本课题的研究内容 1 1.3 文献综述 1 1.3.1 木质素的生物降解 1 1.3.2 木质素降解菌 2 1.3.3 国内外研究概况 2 1.3.4 木质素降解酶及其作用机理 3 1.3.5 微生物产木质素降解酶的影响因素 4 1.3.6 木质素降解酶的应用 5 第二章 实验部分 5 2.1 实验仪器 5 2.2 实验试剂及材料 6 2.3 实验内容与步骤（第一部分） 6 2.3.1 菌种土壤的采集 6 2.3.2 培养基的制备 6 2.3.3 细菌的初步纯种分离 8 2.3.4 细菌的纯种分离 8 2.3.5 纯种