吴珊论文-4.docVIP

本科毕业论文 题目： 纤维素降解菌的分离 学 院: 化学工程与技术学院 专 业: 生物工程 学 号: 200501153042 学生姓名: 吴珊 指导教师: 陈俊 日 期: 2009年6月10日 摘 要 纤维素是地球上最丰富的可再生有机资源，因为难分解大部分未被人类利用。另外，纤维素是造纸废水的COD和SS的主要来源之一。分解纤维素并将其转化成动物易吸收或利用的能源、食物、饲料或化工原料，是纤维素合理应用的重要途径。筛选高效纤维素分解菌，确定其酶学性质是降解纤维素的关键。 本研究从不同环境的土壤中取样，先经过富集培养，然后用纤维素选择琼脂培养基分离菌种，再将单一菌落接种于复筛培养基上，经过刚果红染色，选取在平板上生长快、透明水解圈大的为纤维素降解菌。将此菌株接入滤纸崩解培养基中，观察滤纸降解情况及溃烂速度，比较各株纤维素降解菌降解纤维素的能力。将此菌株制成菌悬液后，接种于液体摇瓶培养基中，取样测定CMC酶活性、并测定滤纸酶活，筛选出酶活力及残余还原糖浓度高的菌株并研究其特性。实验结果表明： （1）实验所取的不同环境的土样中，均有纤维素降解菌的存在。经过富集培养和初筛，分离活性较高的纤维素降解菌26株。 （2）经过刚果红染色鉴定，筛选到10株水解圈较大的纤维素降解菌。 （3）以选取的菌株做滤纸崩解实验，对滤纸均有不同程度的降解。 （4）以选取的菌株测定CMC酶活性，菌株c、E和1的CMC酶活相对较高。 （5）以选取的菌株测定滤纸酶活，菌株c与F的滤纸酶活相对较高。 从刚果红平板水解圈的大小、CMC酶活性和滤纸酶活三个指标判断，菌株c具有较高的纤维素酶活性。这为下一步纤维素酶基因的克隆和基因工程菌的构建打下了基础。 关键词： 纤维素降解菌； 刚果红染色； CMC酶活； 滤纸酶活 Abstract Cellulose is the most abundant earth organic renewable resources, but most of it has not been used by us because it is difficult to break down. Also, cellulose is one of the major sources of paper-making waste-water COD and SS. It is an important way to make use of cellulose that cellulose is decomposed and transformed into energy, food, feed or chemical raw materials which are more easily absorbed or used by animal. Screening high activity cellulose-decomposing bacteria is the key factor to degrade cellulose. This study was disigned to sample from different soil under kraurotic branches and putrescent leaves, separate the strain by cellulose agar selection, follow by congo red staining single colony. Single-colonies of fast-growing, big transparent hydrolysis circle were chosen as the cellulose degration bacteria. Inoculating the screened strains on collapse of the medium filter paper culture medium, degradation extent and speed of filter paper was observed and compared. Preparing the screened bacteria suspension and inoculating in liquid medium, enzyme activation of CMC and filter paper were measured. High efficient strains could be selected by the residual