本文選題：微生物吸附　切入點(diǎn)：納米磁珠　出處：《哈爾濱工業(yè)大學(xué)》2015年碩士論文

【摘要】：微生物在環(huán)境治理中發(fā)揮著重要的作用,在環(huán)境微生物技術(shù)中存在各種形式的界面反應(yīng),影響微生物在介質(zhì)表面的貼附。研究微生物與金屬氧化物的界面反應(yīng),有助于解決膜污染、生物腐蝕等一系列的環(huán)境問題,也可指導(dǎo)開發(fā)新型微生物選擇性富集分離技術(shù),因而在微生物檢測以及環(huán)境風(fēng)險(xiǎn)預(yù)警方面都具有重要的意義。本研究基于DLVO理論指導(dǎo),開發(fā)納米磁珠分離技術(shù),實(shí)現(xiàn)對玻璃纖維濾膜上微生物的分離;結(jié)合經(jīng)典過濾理論與鐵載體在金屬氧化物表面形成化學(xué)鍵的新型作用形式,表征鐵載體介導(dǎo)下,微生物吸附行為特征;開發(fā)鐵載體修飾的磁性材料,探究修飾后材料選擇性富集特定微生物的可行性,取得如下研究成果:納米磁珠分離技術(shù)在酸性條件下,吸附玻璃纖維濾膜上的微生物,具有最大的收率;經(jīng)16sRNA基因高通量測序及群落分析,酸性條件下磁珠法富集微生物的群落結(jié)構(gòu)與原有微生物組成高度近似。共線性分析顯示酸性條件下吸附微生物樣品與對照樣品間的夾角(16.2°)與同一微生物群落DNA兩次PCR擴(kuò)增的結(jié)果近似(14.2°),顯示該方法能夠?qū)崿F(xiàn)對濾膜上微生物無差別的吸附。構(gòu)建了基于濾柱實(shí)驗(yàn)的微生物吸附表征體系。胞外聚合物(Extracellular Polymeric Substances,EPS)缺失型菌株EPS-與野生型具有EPS菌株EPS+在玻璃珠濾料中,表現(xiàn)出不同的過濾形式。EPS-符合經(jīng)典過濾理論的log-linear模型。而EPS+由于EPS影響,表現(xiàn)出與經(jīng)典過濾模型相偏差的形式,即在濾層15 cm位置出現(xiàn)最大截留濃度的顯著拐點(diǎn)。在甲磺酸去鐵胺(Deferoxamine B,DFOB)修飾濾料實(shí)驗(yàn)中,BW25113表現(xiàn)為與經(jīng)典過濾理論相偏差。濾料中截留細(xì)菌濃度先增后減,在濾層15 cm處能截留最大的細(xì)菌濃度。作為對照,在未用DFOB浸泡過的濾料中,由于EPS的存在截留細(xì)菌也表現(xiàn)為先增后減,在濾層下10 cm具有最大的截留細(xì)菌量。相比較對照組,實(shí)驗(yàn)組截留BW25113的細(xì)菌量顯著增高了50%。制備的鐵載體修飾的磁性納米材料,在編碼去鐵胺受體蛋白過表達(dá)菌fhuE++和編碼去鐵胺受體蛋白缺失型菌fhuE-組成的混合體系中,鐵載體修飾后的磁珠能夠優(yōu)先吸附fhuE++菌株,吸附fhuE++的數(shù)量是fhuE-的4倍,且遠(yuǎn)高于裸Fe_3O_4吸附fhuE++的量。同時,甲磺酸去鐵胺修飾后的納米磁珠,在酸性和中性條件下,吸附深圳灣海水中最易與DFOB結(jié)合的放線菌數(shù)量顯著的高于堿性條件和裸Fe_3O_4磁珠吸附的放線菌量。
[Abstract]:Microorganisms play an important role in environmental treatment. There are various forms of interfacial reactions in environmental microbial technology, which affect the adhesion of microbes to the surface of media. The interfacial reactions between microbes and metal oxides are studied. It helps to solve a series of environmental problems, such as membrane fouling, biological corrosion and so on, and can also guide the development of new microorganism selective enrichment and separation technology. Therefore, it is of great significance in microbial detection and environmental risk early warning. Based on the guidance of DLVO theory, the separation technology of nano-magnetic beads is developed to achieve the separation of microbes on glass fiber filter membrane. Combined with the classical filtration theory and the new form of iron carrier forming chemical bond on the surface of metal oxide, the adsorption behavior of microorganism was characterized by iron carrier, and the magnetic material modified by iron carrier was developed. The feasibility of selective enrichment of specific microbes by modified materials was explored. The following results were obtained: under acidic conditions, the nanomagnetic bead separation technology adsorbed microorganism on glass fiber filter membrane with maximum yield; High throughput sequencing and community analysis of 16sRNA gene were performed. The community structure of microbe enriched by magnetic bead method under acidic condition is similar to that of the original microorganism. Co linear analysis shows that the angle between adsorbed microorganism sample and control sample under acid condition is 16.2 擄) and that of the same microbial community DNA. The results of secondary PCR amplification were approximately 14.2 擄, which showed that the method could achieve the same adsorption of microorganism on filter membrane. A microorganism adsorption characterization system based on filter column experiment was constructed. The extracellular Polymeric substancesEPS-deleted strain EPS- and wild strain EPS- were constructed. Type A strain EPS with EPS in glass bead filter media, Different filtering forms. EPS- log-linear model, which accords with the classical filtering theory, and EPS, because of the influence of EPS, shows the form of deviation from the classical filtering model. In the experiment of deferoxamine methanesulfonate modified with DFOB, BW25113 deviated from the classical filtration theory. The concentration of bacteria in filter media increased first and then decreased. The maximum bacterial concentration could be retained in the filter layer of 15 cm. As a control, in the filter media without DFOB immersion, the amount of bacteria interception was increased first and then decreased because of the presence of EPS, and the maximum amount of bacteria interception was found at 10 cm below the filter layer. Compared with the control group, there was no significant difference between the two groups. The amount of bacteria intercepting BW25113 in the experimental group increased significantly by 50%. The magnetic nanomaterials modified with iron carrier were prepared in a mixed system composed of fhuE encoding the deferramine receptor protein overexpression strain and fhuE- encoding the deferramine receptor protein deletion strain. The magnetic beads modified with iron carrier could preferentially adsorb fhuE strain, and the amount of fhuE adsorbed was 4 times as much as that of fhuE-, and was much higher than that of bare Fe_3O_4 adsorbed fhuE. At the same time, in acidic and neutral conditions, the modified magnetic beads were treated with deferoxylamine mesylate. The quantity of actinomycetes most easily bound to DFOB in Shenzhen Bay seawater was significantly higher than that in alkaline conditions and bare Fe_3O_4 beads.
【學(xué)位授予單位】：哈爾濱工業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：X172

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