【作者】 李卓；

【导师】 王继勇； 陶正恒；

【作者基本信息】 武汉理工大学 ， 化学工程， 2012， 硕士

【摘要】 水资源是强国之本,但随着中国工业化进程的推进和中国社会经济的不断发展,我国的环境污染已经非常严重,水资源的状况也越来越令人堪忧。这其中污泥对水资源的影响特别严重,如何合理利用污泥、治理污泥,是当今我国乃至世界的重大课题之一。本文旨在从样品中分离出能高效产生蛋白酶和纤维素酶的高温菌菌株,并通过实验分析和优化高温菌菌株的产酶条件,逐步达到高温菌菌株产酶的最大效率,检测它们的酶活力,评估其应用潜能。其次把分离出的高效产酶的高温菌菌株作为生物添加剂,添加到污泥当中,混合均匀后进行污泥的高温堆肥,通过一个星期左右的堆肥使污泥达到国家农业部有机腐熟料的标准,使其变废为宝。从样本中分离到3株产酶菌株,其细胞均为球形,均能产纤维素酶和蛋白酶,用Al,A2,A3表示,其纤维素酶活分别为21.34U/ml,13.46U/ml,30.77U/ml。其蛋白酶酶活分别为20.44U/ml,23.48U/ml,26.36U/ml。采用单因素实验进行生长条件的优化,分别探索碳源、氮源、温度、pH对A1,A2,A3三种高温菌酶活力的影响。同时得出A1,A2,A3三种高温菌产纤维素酶和蛋白酶的最大值条件。通过单因素实验进行生长条件的优化后,分析A1,A2,A3的数据结果,全面考虑后,A2高温菌的产纤维素酶和蛋白酶的效率高,并把A2高温菌的培养温度定为60℃,培养基pH值定为7.0后进行培养基成分『正交实验,得出结论。高温菌A2的培养基成分为葡萄糖4g/L,羧甲基纤维素钠1lg/L,硝酸铵8g/L,蛋白胨2g/L,硫酸镁0.25g/L、磷酸二氢钾0.5g/L后,高温菌A2生长最旺盛,其产纤维素酶和蛋白酶效率最高。根据国家机肥腐熟剂和卫生指标的合格条件,把高温菌A2进行液体的扩大培养和固体的扩大培养,得到符合国家机肥腐熟剂和卫生指标的规定,固体堆肥温度在55℃以上达到了3天,各处固体堆肥后pH值介于7.2之间,菌数为2.4×1012个,纤维素酶活260.11U/ml,蛋白酶活137.25U/m。把制得的有机腐熟剂作为生物添加剂,添加到污泥当中,与污泥,稻草,麦秆等一起进行高温堆肥,通过一个多星期的高温堆肥后,备处堆肥后的pH值介于7.4之间,菌数为2.6×1011个,纤维索酶活336.67U/ml,蛋白酶活76.67U/ml。更多还原

【Abstract】 The water resources are very important for a country. But with the advance of China’s industrialization process and the continuous development of China’s social-economic. The environmental pollution of China has been very serious, and the status of water resources is more and more concerned, of which the sludge has particularly serious influence on water resources. How to make reasonable use of sludge and practical treatment of sludge is one of the major issues of our country and current world.This paper aims at separating the high temperature bacteria strains which can effectively produce protease and cellulose enzyme from the sample, analyzing and optimizing the high temperature bacteria strains enzyme production conditions through the experimental analysis and, gradually achieving the high temperature bacteria strains of enzyme production’s maximum efficiency, testing their enzyme, and evaluating the potential of application.Secondly add the high temperature bacteria strains with efficient enzyme production capacity which separated from the sample to the sludge as biological additives, make the high-temperature composting after blending. Through a week or so, the sludge reaches the national ministry of agriculture composting of organic compost material standard, then make its reuse.Three strains were isolated from the sample, and they are all the coccus. They were separated for producing cellulase, designated as A1, A2and A3, with enzymatic activity of21.34U/ml,13.46U/ml,30.77U/ml,respectively. They also produce protease, with enzymatic activity of20.44U/ml,23.48U/ml,26.36U/ml, respectively.Respectively explore the optimized parameters for the three strains containing the types of carbon source, nitrogen source, the fermentation temperature and pH to A1, A2, A3three high temperature bacteria the influence of the enzyme activity. And draw A1, A2, A3three high temperature bacteria produce cellulose enzyme and protease maximum conditions.After optimizing the growth condition through single factor experiment, analyze the data of A1, A2, and A3. After the comprehensive consideration, the thermophilic bacterium in A2is the most efficient when producing the cellulase and protease. Then draw the conclusion by taking a medium components of orthogonal experiment after setting the temperature of the high temperature bacteria of A2as60℃, and medium with pH7.0.The composition of the medium of high temperature bacteria in A2includes glucose4g/L, sodium carboxy methyl cellulose11g/L, ammonium nitrate8g/L, specially designed2g/L, magnesium sulfate0.25g/L, potassium dihydrogen phosphate0.5g/L. The A2high-temperature bacteria grows most eugenic, and has the highest efficiency of producing cellulose enzyme and protease.According to the national machine fat rotten agent and health indicators of qualified conditions, expand the culture of A2high-temperature bacteria of liquid and of solid to conform the national standard. Set the temperature of solid compost above55℃for three days; the pH value is between7.2after composting; the bacterium number is2.4x1012; cellulose enzyme activity is260.11U/ml and protease activity is137.25U/m.Add the organic composting agents we made to the sludge as biological additives, then make high temperature composting with sludge and straws together. After high temperature composting for a week, the pH value everywhere after composting is between7.4; the bacterium number is2.6x1011; cellulose enzyme activity is336.67U/ml; protease activity is76.67U/ml.更多还原