潤(rùn)滑油生物降解菌種選育及特性研究
發(fā)布時(shí)間：2018-10-10 17:29
【摘要】：潤(rùn)滑油在工農(nóng)業(yè)生產(chǎn)中有著廣泛的應(yīng)用。由于潤(rùn)滑油回收利用渠道不暢通，回收技術(shù)不成熟，回收費(fèi)用高等因素，導(dǎo)致潤(rùn)滑油的回收利用率低。同時(shí)由于生產(chǎn)工藝不合理以及操作不當(dāng)?shù)韧饨缫蛩貙?dǎo)致大量潤(rùn)滑油流入環(huán)境。這類不可再生潤(rùn)滑油中含有環(huán)烷烴，芳香烴及重金屬等難降解組分，如果不加以有效處理，會(huì)造成環(huán)境污染。物化方法在處理廢潤(rùn)滑油污染時(shí)，，雖然能夠在短時(shí)間里取得一定的效果，但是二次污染嚴(yán)重。而生物法處理廢潤(rùn)滑油是利用微生物的本質(zhì)屬性，即生物降解特性，將有毒害污染物作為底物進(jìn)行徹底降解。作為生物處理技術(shù)的核心，開發(fā)高效降解菌種是目前潤(rùn)滑油污染治理的研究重心。 本實(shí)驗(yàn)旨在從土壤中篩選出具有自主知識(shí)產(chǎn)權(quán)的高效廢潤(rùn)滑油菌種。課題組以廢舊潤(rùn)滑油和油污染土壤為分離源，對(duì)后者首先進(jìn)行預(yù)處理及梯度馴化，再以傳統(tǒng)的微生物分離純化技術(shù)為基礎(chǔ)，分別從廢潤(rùn)滑油和油污染土壤中共分離出51株單菌落；繼而利用24孔板實(shí)驗(yàn)初篩得到15株潤(rùn)滑油降解菌，之后又通過(guò)搖瓶降解實(shí)驗(yàn)和脂肪酶的測(cè)定定量化優(yōu)選出6株高效潤(rùn)滑油降解菌種；最后以這些菌種為物質(zhì)基礎(chǔ)對(duì)其潤(rùn)滑油生物降解能力進(jìn)行了研究。研究證實(shí)這6株菌均具有較好的耐油特性，能夠以潤(rùn)滑油為唯一碳源實(shí)現(xiàn)有效降解，并得到如下結(jié)論： 1）廢潤(rùn)滑油及潤(rùn)滑油長(zhǎng)期污染土壤中都含有高效石油烴降解菌種，是優(yōu)良菌種的理想來(lái)源； 2）利用24孔板實(shí)驗(yàn)，以細(xì)胞培養(yǎng)液和靜止細(xì)胞為接種體，能夠定性的評(píng)價(jià)菌種對(duì)潤(rùn)滑油的生物降解能力，采取該方法篩選出15株目的菌種，其油膜變化情況與孔板培養(yǎng)液中細(xì)胞生長(zhǎng)情況吻合; 3）利用搖瓶降解實(shí)驗(yàn)，并輔以細(xì)胞脂肪酶測(cè)定實(shí)驗(yàn)，篩選出6株降解潤(rùn)滑油優(yōu)勢(shì)菌種，盡管對(duì)6株菌的脂肪酶含量檢測(cè)發(fā)現(xiàn)其中2株菌的脂肪酶含量較低，但在反復(fù)的搖瓶降解實(shí)驗(yàn)中證實(shí)了這2株菌確有較為突出的潤(rùn)滑油降解能力，因此，細(xì)胞脂肪酶測(cè)定只能作為評(píng)價(jià)菌種降解潤(rùn)滑油能力的一個(gè)參考依據(jù)，即有部分菌種代謝產(chǎn)生脂肪酶能力較弱，但不意味著其潤(rùn)滑油生物降解特性一定很差； 4）實(shí)驗(yàn)采用了國(guó)標(biāo)和國(guó)際流行方法對(duì)脂肪酶活力進(jìn)行了檢測(cè)，結(jié)果表明氫氧化鈉滴定法不適合于本實(shí)驗(yàn)體系，表現(xiàn)在對(duì)各株菌的檢測(cè)結(jié)果均較低且酶活力基本都保持一致，而利用紫外分光光度法成功地檢測(cè)了細(xì)胞的脂肪酶活力； 5）通過(guò)對(duì)6株菌的生物降解特性研究發(fā)現(xiàn)，6號(hào)菌種在600mg/L以上的潤(rùn)滑油濃度中不能生長(zhǎng)，但在500mg/L的濃度下，能夠?qū)崿F(xiàn)較高的降解效果。4，5號(hào)菌種在700mg/L的潤(rùn)滑油中也無(wú)法生長(zhǎng)，但在600mg/L濃度以下時(shí)，均具有高效的降解效果。除6號(hào)菌種外，其他5支菌種在144小時(shí)的降解條件下，能夠?qū)崿F(xiàn)對(duì)濃度為500mg/L的潤(rùn)滑油的完全降解。而6號(hào)菌種也能達(dá)到80%左右的降解效率。
[Abstract]:Lubricating oil is widely used in industrial and agricultural production. The recovery and utilization rate of lubricating oil is low due to the lack of smooth recovery channels, immature recovery technology and high recovery cost. At the same time, due to unreasonable production process and improper operation and other external factors leading to a large number of lubricating oil into the environment. This kind of non-renewable lubricating oil contains non-degradable components such as cycloalkanes, aromatics and heavy metals, which will cause environmental pollution if not treated effectively. Although the physicochemical method can obtain certain effect in a short time when dealing with the pollution of waste lubricating oil, the secondary pollution is serious. The biological treatment of waste lubricating oil is based on the essential properties of microorganisms, I. e., biodegradation, which uses toxic pollutants as the substrate for thorough degradation. As the core of biological treatment technology, the development of high-efficiency biodegradable bacteria is the focus of research on the treatment of lubricating oil pollution. The purpose of this study was to screen high efficient waste lubricating oil bacteria with independent intellectual property rights from soil. Using waste lubricating oil and oil-contaminated soil as the source of separation, the research group firstly pretreated and acclimated the latter, and then based on the traditional microbial separation and purification technology. A total of 51 single colonies were isolated from waste lubricating oil and oil-contaminated soil, and 15 strains of degrading bacteria were obtained by 24 hole plate test. After that, six strains of high-efficiency lube oil degrading bacteria were selected by shaking flask degradation experiment and lipase determination. Finally, the biodegradation ability of lubricating oil was studied on the basis of these strains. The results showed that all the 6 strains had good oil tolerance and could be effectively degraded with lubricating oil as the sole carbon source. The conclusions are as follows: 1) waste lubricating oil and lubricating oil contaminated soil for a long time contain high efficient petroleum hydrocarbon degrading bacteria, which is the ideal source of excellent bacteria, 2) using 24 hole plate experiment, The biodegradability of lubricating oil was evaluated qualitatively by using cell culture medium and stationary cells as inoculum, and 15 target strains were screened by this method. The variation of oil film was consistent with that of cell growth in the culture medium. 3) by using shaking flask degradation test and cell lipase assay, 6 strains of dominant bacteria were selected. Although the lipase content of 6 strains was lower than that of 2 strains, it was proved by repeated shaking flask degradation experiments that the two strains had outstanding lube oil degradation ability. The determination of cellular lipase can only be used as a reference for evaluating the ability of bacteria to degrade lubricating oil, that is, the ability of some strains to produce lipase is weak, but it does not mean that the biodegradability of lubricating oil must be very poor. 4) the activity of lipase was detected by national standard and international popular method. The results showed that sodium hydroxide titration was not suitable for the experiment system. The results showed that the detection results of each strain were low and the enzyme activity was basically the same, while the lipase activity of the cells was successfully detected by ultraviolet spectrophotometry. 5) the biodegradation characteristics of 6 strains of bacteria were studied. It was found that strain 6 could not grow in the concentration of lubricating oil above 600mg/L, but at the concentration of 500mg/L. Strain 4 and 5 could not grow in the lubricating oil of 700mg/L, but when the concentration of 600mg/L was below the concentration, they all had high degradation effect. With the exception of strain No. 6, the other five strains could completely degrade the lubricating oil with concentration of 500mg/L under the condition of 144-hour degradation. The degradation efficiency of strain 6 can reach about 80%.
【學(xué)位授予單位】：重慶工商大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類號(hào)】：X172
本文編號(hào)：2262657
[Abstract]:Lubricating oil is widely used in industrial and agricultural production. The recovery and utilization rate of lubricating oil is low due to the lack of smooth recovery channels, immature recovery technology and high recovery cost. At the same time, due to unreasonable production process and improper operation and other external factors leading to a large number of lubricating oil into the environment. This kind of non-renewable lubricating oil contains non-degradable components such as cycloalkanes, aromatics and heavy metals, which will cause environmental pollution if not treated effectively. Although the physicochemical method can obtain certain effect in a short time when dealing with the pollution of waste lubricating oil, the secondary pollution is serious. The biological treatment of waste lubricating oil is based on the essential properties of microorganisms, I. e., biodegradation, which uses toxic pollutants as the substrate for thorough degradation. As the core of biological treatment technology, the development of high-efficiency biodegradable bacteria is the focus of research on the treatment of lubricating oil pollution. The purpose of this study was to screen high efficient waste lubricating oil bacteria with independent intellectual property rights from soil. Using waste lubricating oil and oil-contaminated soil as the source of separation, the research group firstly pretreated and acclimated the latter, and then based on the traditional microbial separation and purification technology. A total of 51 single colonies were isolated from waste lubricating oil and oil-contaminated soil, and 15 strains of degrading bacteria were obtained by 24 hole plate test. After that, six strains of high-efficiency lube oil degrading bacteria were selected by shaking flask degradation experiment and lipase determination. Finally, the biodegradation ability of lubricating oil was studied on the basis of these strains. The results showed that all the 6 strains had good oil tolerance and could be effectively degraded with lubricating oil as the sole carbon source. The conclusions are as follows: 1) waste lubricating oil and lubricating oil contaminated soil for a long time contain high efficient petroleum hydrocarbon degrading bacteria, which is the ideal source of excellent bacteria, 2) using 24 hole plate experiment, The biodegradability of lubricating oil was evaluated qualitatively by using cell culture medium and stationary cells as inoculum, and 15 target strains were screened by this method. The variation of oil film was consistent with that of cell growth in the culture medium. 3) by using shaking flask degradation test and cell lipase assay, 6 strains of dominant bacteria were selected. Although the lipase content of 6 strains was lower than that of 2 strains, it was proved by repeated shaking flask degradation experiments that the two strains had outstanding lube oil degradation ability. The determination of cellular lipase can only be used as a reference for evaluating the ability of bacteria to degrade lubricating oil, that is, the ability of some strains to produce lipase is weak, but it does not mean that the biodegradability of lubricating oil must be very poor. 4) the activity of lipase was detected by national standard and international popular method. The results showed that sodium hydroxide titration was not suitable for the experiment system. The results showed that the detection results of each strain were low and the enzyme activity was basically the same, while the lipase activity of the cells was successfully detected by ultraviolet spectrophotometry. 5) the biodegradation characteristics of 6 strains of bacteria were studied. It was found that strain 6 could not grow in the concentration of lubricating oil above 600mg/L, but at the concentration of 500mg/L. Strain 4 and 5 could not grow in the lubricating oil of 700mg/L, but when the concentration of 600mg/L was below the concentration, they all had high degradation effect. With the exception of strain No. 6, the other five strains could completely degrade the lubricating oil with concentration of 500mg/L under the condition of 144-hour degradation. The degradation efficiency of strain 6 can reach about 80%.
【學(xué)位授予單位】：重慶工商大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類號(hào)】：X172
【參考文獻(xiàn)】
相關(guān)期刊論文 前5條
1 沈齊英;;可生物降解潤(rùn)滑劑的研究概況[J];北京石油化工學(xué)院學(xué)報(bào);2011年02期
2 王昌祿;趙璇;王玉榮;孟祥云;;蓖麻基潤(rùn)滑油降解菌群的構(gòu)建及對(duì)土壤的修復(fù)作用[J];河北工業(yè)大學(xué)學(xué)報(bào);2011年01期
3 張楠;陳波水;楊致邦;余瑛;黃偉九;;一株潤(rùn)滑油降解放線菌的分離鑒定及特性[J];石油學(xué)報(bào)(石油加工);2010年06期
4 張楠;陳波水;余瑛;黃偉九;方建華;;潤(rùn)滑油降解菌群的構(gòu)建及其降解性能[J];石油學(xué)報(bào)(石油加工);2010年S1期
5 劉今金,董延庭;生物可降解潤(rùn)滑油[J];河南石油;2005年05期
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