本文選題：錳氧化細(xì)菌 + 分離鑒定�。� 參考：《哈爾濱工業(yè)大學(xué)》2014年碩士論文

【摘要】：錳在地殼中的含量僅此于鐵，是地殼中的主要成分。廣泛存在于自然界中，是人體所必需的微量元素之一，但是如果攝入錳量過多會引起神經(jīng)性疾病，危害身體健康。我國飲用水要求規(guī)定：對錳含量的最高允許濃度為0.1mg/L。且在生活用水和工業(yè)用水過程中，錳含量超標(biāo)也是影響水質(zhì)的主要因素之一，如影響水的色度和氣味等。近年來，，生物除錳工藝得到了發(fā)展和廣泛應(yīng)用，與傳統(tǒng)化學(xué)除錳法相比，生物氧化除錳具有處理效果好，處理效果穩(wěn)定，運(yùn)行所需費(fèi)用低等明顯優(yōu)勢。水相和土壤環(huán)境中廣泛存在著錳氧化微生物，可高效地將Mn(II)氧化成Mn(IV)，在錳的生物地球化學(xué)循環(huán)中具有重要的作用。有些錳氧化細(xì)菌具有極強(qiáng)的Mn(IV)的生成率，比化學(xué)催化快10萬倍左右。因此，錳氧化細(xì)菌在生物除錳研究中具有較高的應(yīng)用價(jià)值，可用于含錳量高的水處理和土壤中重金屬污染的治理。但是，目前對于錳氧化菌除錳機(jī)制的研究還不夠深入。 本實(shí)驗(yàn)利用三種不同富集培養(yǎng)基對不同環(huán)境樣品進(jìn)行富集，通過LBB指示劑法篩選富集效果好的樣品作為分離樣品。同時(shí)利用LBB指示劑法篩選具有錳氧化能力的菌株，待分離純化后通過生理生化及分子生物學(xué)等鑒定方法進(jìn)行分析鑒定。并測定分離菌株生長曲線、及接種量、Mn2+濃度、pH值等對生長曲線的影響及研究分離菌株的錳氧化特性。 本研究以土壤和河流底泥等為分離樣品，首先，利用PYCM培養(yǎng)基、K培養(yǎng)基和PC培養(yǎng)基進(jìn)行富集和分離。對分離獲得的10株細(xì)菌通過LBB指示劑進(jìn)行了對Mn(II)的氧化能力檢測。然后通過MIDI微生物鑒定系統(tǒng)、Biolog分析、生理生化分析和16S rDNA測序分析，初步鑒定菌種MB1為地衣芽胞桿菌（Bacillus licheniformis），MB2為巨大芽孢桿菌（Bacillus-megaterium）。 實(shí)驗(yàn)結(jié)果表明不同接種量對除錳菌MB1和MB2的生長曲線總體來說影響不大。MB1的最適接種量為5%，MB2的最適接種量為15%。當(dāng)培養(yǎng)基的pH值為5.0-7.0時(shí)，MB1和MB2菌體均在此范圍內(nèi)可以生長的很好，當(dāng)pH值為7.0時(shí)，菌體生長狀態(tài)達(dá)到最好，培養(yǎng)基中生物量最大，說明在此條件下菌體能夠很快適應(yīng)環(huán)境快速進(jìn)入對數(shù)生長期，且對數(shù)期時(shí)間較其它pH值條件下菌體對數(shù)期時(shí)間更長。之后，隨著培養(yǎng)基堿性增強(qiáng)，生物量又逐漸減小，直至pH值為9.0時(shí)菌體的生長被抑制。當(dāng)pH值為9.0時(shí)，堿性環(huán)境中錳氧化菌的生理活動(dòng)受到影響。在pH值為6.0-8.0的條件下，MB1和MB2對Mn2+氧化率均在35%以上。對除錳菌的耐錳能力進(jìn)行分析，研究了不同Mn2+濃度對菌體生長的影響，當(dāng)濃度高于12mmol/L時(shí)，MB1菌體的生長速度很慢甚至幾乎不生長，表明當(dāng)Mn2+濃度過高時(shí)會抑制細(xì)菌的生長。MB2菌株在含Mn2+培養(yǎng)基中的菌量比不含Mn2+的小，但當(dāng)培養(yǎng)基在12mmol/L范圍以內(nèi)變化差距不大，表明MB2菌對高濃度Mn2+的耐受性比MB1菌要好。 通過原子吸收法測定菌株除錳率，在K培養(yǎng)基中培養(yǎng)7天后，菌種MB1的錳氧化率為61.7%，菌種MB2的錳氧化率為90%。研究初始pH值（5-9）的K培養(yǎng)基中，pH值對錳氧化率的影響，在pH值為7.0時(shí)，MB1和MB2的生物氧化率最高，可達(dá)到80%左右。當(dāng)pH為5.0和6.0時(shí)，菌株MB1和MB2對Mn2+氧化率不到60%。說明酸性環(huán)境影響了錳氧化菌的生長，可能是由于酸性環(huán)境使培養(yǎng)基的組成發(fā)生改變，從而導(dǎo)致錳氧化率下降。通過研究錳氧化菌在生長過程中對pH值的影響表明錳氧化菌使培養(yǎng)基pH值升高。對除錳菌MB1和MB2的除錳特性進(jìn)行分析，證明本實(shí)驗(yàn)中所篩選到的錳氧化細(xì)菌在對Mn2+進(jìn)行氧化過程中包含生物氧化和化學(xué)氧化的共同作用。 該菌為進(jìn)一步研究錳氧化機(jī)制和實(shí)現(xiàn)強(qiáng)化除錳提供了重要的菌種。
[Abstract]:Manganese content in the crust is only iron, it is the main component of the earth's crust. It is widely found in nature and is one of the essential trace elements necessary for the human body. However, excessive intake of manganese can cause neurogenic diseases and harm the health of the body. The requirement for drinking water in our country is that the maximum allowable concentration of manganese content is 0.1mg/L. and in domestic water use. In the process of water use, manganese content is also one of the main factors affecting the water quality, such as the color and odor of water. In recent years, the biological manganese removal process has been developed and widely used. Compared with the traditional chemical manganese removal method, biological oxidation removal of manganese has the advantages of good treatment effect, stable treatment effect and low cost of operation. Manganese oxide microorganism is widely used in water and soil environment, which can efficiently oxidize Mn (II) into Mn (IV) and play an important role in the biogeochemical cycle of manganese. Some manganese oxidizing bacteria have a very strong Mn (IV) formation rate, 100 thousand times faster than chemical catalysis. Therefore, manganese oxide bacteria have a better effect in the study of biological manganese removal. High application value can be used in the treatment of high manganese content water and the treatment of heavy metal pollution in soil. However, the research on manganese removal mechanism of manganese oxidizing bacteria is not enough.
In this experiment, three different enrichment medium were used to enrich the samples of different environment. The samples with good effect were screened by LBB indicator method. At the same time, the strains with manganese oxidation ability were screened by LBB indicator method. After separation and purification, the samples were identified by physiological and biochemical and molecular biology identification methods. The effects of growth curve, inoculum size, Mn2+ concentration and pH value on the growth curve and the manganese oxidation characteristics of the isolates were also determined.
In this study, the samples were separated from soil and river sediment. First, the PYCM medium, K medium and PC medium were enriched and separated. The oxidation capacity of Mn (II) was detected by the LBB indicator for 10 strains of isolated bacteria. Then, the MIDI microbial identification system, Biolog analysis, physiological and biochemical analysis and 16S rDNA sequencing were carried out. Preliminary analysis showed that the strain MB1 was Bacillus licheniformis (Bacillus licheniformis) and MB2 was Bacillus MEG (Bacillus-megaterium).
The experimental results showed that the optimum inoculation for the growth curve of manganese bacteria MB1 and MB2 was 5%, and the optimum inoculation of MB2 was 15%. when the pH value of the medium was 5.0-7.0, MB1 and MB2 could grow well in this range. When the pH value was 7, the growth state of the bacteria reached the best, in the medium. The biomass is the largest, indicating that the bacteria can quickly adapt to the environment and quickly enter the logarithmic growth period, and the logarithmic time is longer than the logarithmic period of the mycelium under the other pH values. Then, the biomass is gradually reduced as the base of the medium increases, and the growth of the bacteria is inhibited when the pH value is 9. When the value of pH is 9, the alkaline environment is in the alkaline environment. The physiological activities of the medium manganese oxidizing bacteria were affected. Under the condition of pH 6.0-8.0, the oxidation rate of Mn2+ and MB1 was above 35%. The manganese resistance of manganese removal bacteria was analyzed. The effect of different Mn2+ concentration on the growth of the bacteria was studied. When the concentration was higher than 12mmol/L, the growth rate of MB1 bacteria was slow or almost no growth, indicating that when Mn2+ is Mn2+. When the concentration was too high, the bacterial growth of.MB2 strain in the Mn2+ medium was smaller than that of Mn2+, but the gap in the medium of 12mmol/L was little, indicating that the tolerance of MB2 bacteria to high concentration Mn2+ was better than that of MB1 bacteria.
The manganese removal rate of strain was measured by atomic absorption spectrometry. The manganese oxidation rate of the strain MB1 was 61.7% after 7 days in the K culture medium. The manganese oxidation rate of the strain MB2 was 90%. in the initial pH value (5-9) K medium. The effect of pH value on the oxidation rate of manganese was 7, and the oxidation rate of MB1 and MB2 was up to 80% when pH value was 7. When pH was 5 and 6, The oxidation rate of Mn2+ by strain MB1 and MB2 indicates that the oxidation rate of Mn2+ affects the growth of manganese oxide bacteria, which may be due to the change of the composition of the medium in acid environment and the decrease of manganese oxidation rate. The effect of Manganese Oxidizing Bacteria on the pH value in the growth process indicates that manganese oxidizing bacteria increase the medium pH value of the manganese oxide bacteria and the manganese removing bacteria MB1. The manganese removal characteristics of MB2 have been analyzed. It is proved that the manganese oxide bacteria screened in this experiment include the common effect of biological oxidation and chemical oxidation during the oxidation of Mn2+.
This strain provides an important strain for further study of manganese oxidation mechanism and enhanced manganese removal.

【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】：2014
【分類號】：TU991.2;X172

【參考文獻(xiàn)】

相關(guān)博士學(xué)位論文 前1條

1 曾輝平;含高濃度鐵錳及氨氮的地下水生物凈化效能與工程應(yīng)用研究[D];哈爾濱工業(yè)大學(xué);2010年

相關(guān)碩士學(xué)位論文 前6條

1 鄭金;一株錳氧化菌對Mn~(2+)的氧化及其產(chǎn)物的表征和特性研究[D];華中農(nóng)業(yè)大學(xué);2013年

2 鐘律;生物法過濾處理模擬含鐵錳地下水試驗(yàn)研究[D];華東交通大學(xué);2012年

3 唐美麗;錳氧化蛋白的研究[D];北京工業(yè)大學(xué);2011年

4 季虹;生物氧化錳的形成及其對典型環(huán)境雌激素的氧化降解[D];浙江工業(yè)大學(xué);2011年

5 歐亞西;錳氧化細(xì)菌在粘土礦物表面的吸附及其對Mn（Ⅱ）的氧化[D];華中農(nóng)業(yè)大學(xué);2009年

6 姚遠(yuǎn);富鐵錳地下水的微生物治理[D];福建師范大學(xué);2009年

本文編號：1813071