成都理工大学毕业设计I氧化亚铁硫杆菌的分离及生物学特性研究作者姓名:冯雷专业:生物工程指导老师:谢鸿观摘要氧化亚铁硫杆菌(Thiobacillusferrooxidans,简称T.f)属革兰氏阴性无机化能自养菌、嗜酸、专性好氧,靠氧化基质中的Fe2+为Fe3+和低价态硫为硫酸根而获取能量。它广泛存在于土壤海水、淡水、垃圾、硫磺泉和沉积硫内,尤以金属硫化矿和煤矿等酸性矿坑水中最为常见。因此,本论文从T.f菌的分离纯化出发,进行了T.f菌的生理生化及其生物活性的研究。本文从浸矿溶液中分离得到的氧化亚铁硫杆菌进行研究,结果表明硫酸亚铁相对于硫来说是一种速效能源,单质硫是长效能源,并且能提供比亚铁更高的能量。关键词:氧化亚铁硫杆菌,微生物冶金,生物浸矿,分离纯化成都理工大学毕业设计IIIsolationofThiobacillusandbiologicalcharacteristicsName:FengLeiMajor:BioengineeringSupervisor:XieHongguanAbstractFerrooxidans(Thiobacillusferrooxidans,calledT.f),aninorganicchemoautotrophGramnegativebacteria,acidophilus,specificaerobic,relyingonsubstrateoxidationinFe3+andFe2+forthelowstateofsulfurtosulfuricacidrootsandobtainenergy,whichiswidelyfoundinsoilwater,freshwater,waste,anddepositionofsulfurinsulfursprings,especiallyinsulfidemineralssuchasacidminewaterandcoalthemostcommon.Therefore,thispaperfromtheT.fpurificationandidentificationofbacteriainconductingthephysiologicalstrainAfitsbiologicalactivity.Thisseparationfromtheleachingsolutionobtainedferrooxidansstudyresultsshowthatcomparedwithferroussulfatesulfurisakindofquickenergy,sulfurisalong-lastingenergy,andcanprovidemoreenergythantheferrous.Keywords:Thiobacillusferrooxidans,microbioleaehin,Biologicalleaching,isolation成都理工大学毕业设计III目录摘要.....................................IAbstract................................II一、前言............................11.1生物冶金的概念............................................................................................................................11.2碲矿的生物冶金...........................................................................................................................21.3氧化亚铁硫杆菌的作用................................................................................................................31.4本实验研究内容...........................................................................................................................4二、实验部分.............................52.1实验材料与设备............................................................................................................................52.1.1菌种采集.................................................................................................................................52.1.2实验试剂.................................................................................................................................52.1.3实验仪器.................................................................................................................................52.1.4培养基.....................................................................................................................................62.2研究方法........................................................................................................................................72.2.1菌种富集培养.........................................................................................................................72.2.2分离纯化.................................................................................................................................72.2.3单层平板的制作.....................................................................................................................82.2.4双层平板的制作.....................................................................................................................92.2.5革兰氏染色.............................................................................................................................92.2.6pH测定................................................................................................................................92.2.7细菌计数方法.........................................................................................................................92.4生物特性研究..............................................................................................................................112.4.1生长曲线...............................................................................................................................112.4.2氧化亚铁硫杆菌菌株对能源的利用情况...........................................................................112.4.3最适生长温度的测定...........................................................................................................112.4.4最适初始pH值的测定........................................................................................................11结果与讨论..............................12结论....................................15致谢....................................16参考文献................................17成都理工大学毕业设计1一、前言矿物质是在工业生产中不可缺少的物质,是经济建设的重要物质基础。我国含有的矿产资源种类丰富,矿产已探明储量丰富。我国是一个地大物博的强大国家,但由于人口基数大,导至各种资源人均占有量低于世界平均水平。所以我国矿产资源相对紧缺,富矿多数已开发利用,还有很多贫瘠矿,矿含量相对低于其它富矿,开采难度大。现有开采技术的开采方式对环境的破坏程度大,提取矿物质也不完全正确,很多矿仍不能开采。针对目前的情况,微生物浸矿是一种新的开采方式,具有溶矿率高,不污染环境等独特优势。1.1生物冶金的概念生物冶金是利用矿物为营养基质的微生物的作用,将矿物中金属溶解,进分离、富集、纯化而提取金属的湿法冶金技术。该技术具有流程短、成本低、环境友好和低污染等特点,特别适于处理贫矿、废矿、表外矿及难采、难选、难冶矿的堆浸和就地浸出。从文献记载来看,中国是世界上最早采用生物冶金技术的国家,早在公元前六、七世纪,就在采铜、铁过程中不自觉地利用了自发生长的某些自养细菌。而在欧洲,这种技术的应用至少始于公元二世纪,从1687年开始,瑞典中部Falun矿山的铜矿至少已经浸出了2百万吨铜[1]。在国外,铜,铀的生物提取以及含砷金矿的预氧化已实现产业化。在铜的