固定化原油降解菌的制备及其降解特性研究

发布时间：2018-04-13 00:24

本文选题：原油 + 生物降解　；参考：《吉林农业大学》2015年硕士论文

【摘要】：随着我国经济的迅猛发展,原油造成的环境污染问题日益严重,同时对原油污染处理技术的研究不断深入,人们发现生物修复处理技术对原油降解效果优异,而且该技术具有操作简单、成本低、对环境无二次污染等特点。本试验从长期受原油污染的土壤中筛选获得2株高效原油降解菌,并制备成固定化菌剂,通过对固定化方法选择及微生物降解特性的研究,最终获得最佳固定化方法及降解条件,并将该方法及条件应用于实验室内原油污染土壤的生物修复中。本试验研究结果如下:1.本试验从松原长期受原油污染的土壤中分离、筛选出2株原油降解菌JL8和JL10。通过形态观察、生理生化试验和16S rDNA分子生物学鉴定,确定原油降解菌JL8为恶臭假单胞菌(Pseudomonas putida sp.)、JL10为铜绿假单胞菌(Pseudomonas aeruqinosa sp.),且原油降解菌JL8和JL10均为产表面活性剂菌株。2.以原油降解菌JL10为研究对象,采用海藻酸钠、聚乙烯醇及海藻酸钠+聚乙烯醇三种固定化方法制备固定化菌剂,用于原油降解试验。通过考察固定化小球的性能及其对原油降解率,初步确定选择聚乙烯醇(10.0%)+海藻酸钠(1.0%)的方法固定化原油降解菌,其成球性、机械性、传质性良好,且对原油的降解率高达81.77%。3.对初步确定的固定化方法进行改良,分别为聚乙烯醇(10.0%)+海藻酸钠(1.0%)+硅藻土和聚乙烯醇(10.0%)+海藻酸钠(1.0%)+活性炭两种固定化方法,并以原油降解率为目标对其进行优化。试验结果表明,最佳固定化方法为聚乙烯醇(10.0%)+海藻酸钠(1.0%)+硅藻土(10g/L)与原油降解菌(OD600=0.75)等体积混合,滴入含1.0%CaCl2的饱和硼酸中,固定化30h。4.以原油降解菌JL8和JL10为固定化目标,对其降解特性进行优化。试验结果表明,原油降解菌JL8在原油浓度10.0g/L、添加量为1.0ml菌悬液制备固定化小球、初始pH为8.0、盐浓度为2.0%条件下,原油降解效果最好;原油降解菌JL10在原油浓度为10.0g/L、添加0.5ml菌悬液制备固定化小球、初始pH为9.0、盐浓度为2.0%条件下,原油降解效果最好。5.采用玉米秸秆粉末15g、硅藻土5g作为吸附剂,分别固定原油降解菌JL8和JL10,并用于原油污染土壤试验室模拟试验中。试验结果表明,经75天生物降解后,固定化JL8和JL10对原油的降解率分别为95.98%和87.20%。综上所述,本研究制备的固定化菌剂对原油的降解率较高,为日后原油污染土壤的生物修复提供了一定的物质基础。
[Abstract]:With the rapid development of economy in China, the environmental pollution caused by crude oil is becoming more and more serious. At the same time, it is found that bioremediation technology has excellent degradation effect on crude oil.Moreover, the technology has the characteristics of simple operation, low cost and no secondary pollution to the environment.In this experiment, two strains of high efficient bacteria were obtained from soil contaminated by crude oil for a long time, and immobilized bacteria were prepared. The selection of immobilization methods and the characteristics of microbial degradation were studied.Finally, the best immobilization method and degradation conditions were obtained and applied to the bioremediation of crude oil contaminated soil in laboratory.The results of this experiment are as follows: 1: 1.Two strains of crude oil degrading bacteria JL8 and JL10 were isolated from soil contaminated by crude oil in Songyuan for a long time.By morphological observation, physiological and biochemical tests and molecular biological identification of 16s rDNA, the crude oil degrading bacteria JL8 was identified as Pseudomonas putida sp. JL10 as Pseudomonas aeruqinosa sp.0.The crude oil degrading bacteria JL8 and JL10 were both surfactant producing strains.The immobilized bacteria agent was prepared by three immobilization methods, sodium alginate, polyvinyl alcohol and sodium alginate polyvinyl alcohol, and used in crude oil degradation test.By investigating the properties of immobilized pellets and their degradation rate of crude oil, it was preliminarily determined that the method of immobilized crude oil degradation bacteria was polyvinyl alcohol 10.0) sodium alginate 1.0). Its spherical, mechanical and mass transfer properties were good, and the degradation rate of crude oil was as high as 81.770.3.The immobilization methods were modified as follows: polyvinyl alcohol 10.0) sodium alginate 1.0) diatomite and polyvinyl alcohol 10.0) sodium alginate 1.0) activated carbon, and the degradation rate of crude oil was optimized.The results showed that the optimum immobilization method was as follows: polyvinyl alcohol 10.0) sodium alginate 1.0) diatomite 10g / L) mixed with crude oil degrading bacteria OD6000.75), dripped into saturated boric acid containing 1.0Cl2, immobilized for 30h 路4.The degradation characteristics of crude oil degrading bacteria JL8 and JL10 were optimized.The results showed that when the crude oil concentration of JL8 was 10.0 g / L, the amount of 1.0ml suspension was added to prepare the immobilized pellets, the crude oil degradation efficiency was the best when the initial pH was 8.0 and the salt concentration was 2.0%.When the crude oil concentration of JL10 was 10.0 g / L, the immobilized pellets were prepared by adding 0.5ml bacteria suspension, and the initial pH was 9.0% and the salt concentration was 2.0%, the degradation effect of crude oil was the best.Corn straw powder 15g and diatomite 5g were used as adsorbent to immobilize crude oil degrading bacteria JL8 and JL10 respectively.The results showed that after 75 days of biodegradation, the degradation rates of crude oil by immobilized JL8 and JL10 were 95.98% and 87.20%, respectively.To sum up, the immobilized bacteria prepared in this study has a high degradation rate of crude oil, which provides a certain material basis for the bioremediation of crude oil contaminated soil in the future.
【学位授予单位】：吉林农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：X172

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