水溶液介質(zhì)中氯酚類有機(jī)污染物的生物修復(fù)研究
發(fā)布時間：2018-11-07 11:57
【摘要】：白腐菌降解污染物具有非專一性、高效性、廣譜性等特點(diǎn),在氯酚類難降解有機(jī)物的生物修復(fù)中有良好的應(yīng)用前景。白腐菌和漆酶在工業(yè)廢水處理和紙漿生物漂白等方面表現(xiàn)出很大的研究價值和應(yīng)用潛力,然而漆酶的生產(chǎn)過程中普遍存在漆酶產(chǎn)量低、生產(chǎn)成本高等問題。本論文通過研究添加廉價天然底物和金屬離子,優(yōu)化產(chǎn)酶培養(yǎng)基,提高漆酶產(chǎn)量,并對各個影響因子進(jìn)行優(yōu)化,結(jié)合發(fā)酵動力學(xué)模型探討白腐菌產(chǎn)酶規(guī)律。分別將白腐菌和漆酶用于氯酚類化合物廢水中,研究白腐菌和漆酶對廢水的生物修復(fù)機(jī)理。旨在為白腐菌和漆酶在廢水處理領(lǐng)域的應(yīng)用提供一定的理論依據(jù)與技術(shù)支持。本論文以糙皮側(cè)耳(Pleurotus ostreatus)為研究對象,通過添加不同天然底物,對糙皮側(cè)耳發(fā)酵產(chǎn)漆酶條件進(jìn)行優(yōu)化。結(jié)果顯示,添加桔皮培養(yǎng)基優(yōu)化效果最好,最高酶活由5U/mL提高為45U/mL,發(fā)酵時間比原來縮短了1/3。同時對其發(fā)酵動力學(xué)進(jìn)行研究,得出A、B兩組動力學(xué)模型,對比A(添加桔皮)、B(對照)兩組發(fā)酵動力學(xué)模型,加入桔皮后,菌體的增殖能力和產(chǎn)漆酶能力都得到大幅度提高,菌體生長速率和產(chǎn)率系數(shù)也得到不同程度的提高。對白腐菌修復(fù)氯酚類廢水的降解機(jī)理進(jìn)行研究,研究發(fā)現(xiàn):P.ostreatus對氯酚/苯酚類物質(zhì)吸附能力順序?yàn)?2,4,6-TCP2-CP苯酚PCP;不同氯酚加入方式并沒有最終引起生物量發(fā)生很大變化,第5d加入2-CP和2,4,6-TCP組酶活分別是逐步加入2-CP和2,4,6-TCP組酶活的10倍和5倍,第5d集中加入氯酚后,三種氯酚去除率相比之前都得到不同程度提高,由此可以通過改變氯酚加入方式的方法來減小氯酚對漆酶酶活的抑制作用,同時可以提高氯酚去除率。研究P.ostreatus去除氯酚的作用分析,氯酚濃度介于10~30mg/L時,2-CP降解作用占絕大部分比例,吸附作用所占比例較小;TCP降解過程中,吸附作用和降解作用所占比例相當(dāng);相對于吸附作用來說,PCP降解作用占大部分比例,降解作用比例隨著濃度增加有所減少。初步純化獲得粗酶,研究了粗酶的性質(zhì):最適反應(yīng)溫度為45℃,最適反應(yīng)pH=3;粗酶降解氯酚過程中,可以通過增加漆酶濃度,利用緩沖溶液,加入介體ABTS(2,2-聯(lián)氨-二(3-乙基苯并噻唑-6-磺酸)二銨鹽),提高氯酚去除率,同時隨著反應(yīng)時間的延長,漆酶酶活逐漸降低;漆酶降解氯酚的動力學(xué)曲線符合一級反應(yīng)動力學(xué)規(guī)律,并根據(jù)阿倫尼烏斯公式,計算得出漆酶催化2-CP,2,4,6-TCP和PCP三種氯酚反應(yīng)的活化能依次為:26.52,33.52和48.19KJ/mol。
[Abstract]:The degradation of pollutants by white rot fungi has the characteristics of non-specificity, high efficiency and broad spectrum, so it has a good application prospect in the bioremediation of chlorophenol refractory organic compounds. White rot fungi and laccase have great research value and application potential in industrial wastewater treatment and pulp biological bleaching. However, laccase production is generally low in production process, high production cost and other problems. In this paper, by adding cheap natural substrates and metal ions, the enzyme production medium was optimized to increase the laccase yield, and various factors were optimized, and the enzyme production law of white rot fungi was discussed based on the fermentation kinetics model. White rot fungi and laccase were used in chlorophenol wastewater respectively to study the bioremediation mechanism of white rot fungi and laccase. The aim is to provide some theoretical basis and technical support for the application of white rot fungi and laccase in wastewater treatment. In this paper, the fermentation conditions of laccase from Pleurotus strigella (Pleurotus ostreatus) were optimized by adding different natural substrates. The results showed that the optimum culture medium of orange peel was the best, the highest enzyme activity was increased from 5U/mL to 45 U / mL, and the fermentation time was shortened by 1 / 3. At the same time, the fermentation kinetics was studied, and the two groups of dynamic models were obtained. Compared with A (orange peel), B (control), two groups of fermentation kinetics models were added, after the addition of orange peel, The ability of cell proliferation and laccase production were greatly improved, and the growth rate and yield coefficient were also improved to some extent. The degradation mechanism of chlorophenol wastewater remediated by white rot fungi was studied. It was found that the adsorption ability of P.ostreatus for chlorophenol / phenol was in the order of 2N 46-TCP2-CP phenol PCP;. Different chlorophenol addition methods did not result in a significant change in biomass. The enzyme activities in the 2-CP and 2H4 + 6-TCP groups on the 5th day were 10 times and 5 times higher than those in the gradually added 2-CP and 2H4 + 6-TCP groups, respectively, and were concentrated on the 5th day after chlorophenol was added. The three chlorophenol removal rates were improved in different degrees, so the inhibition of chlorophenol on laccase activity and the removal rate of chlorophenol could be reduced by changing the method of chlorophenol addition. The effect of P.ostreatus on the removal of chlorophenol was studied. When the concentration of chlorophenol was in the range of 10~30mg/L, the degradation of 2-CP accounted for the vast majority and the proportion of adsorption was small, and the proportion of adsorption and degradation was the same in the process of TCP degradation. Compared with adsorption, PCP degradation accounted for most of the proportion, and the degradation ratio decreased with the increase of concentration. The properties of crude enzyme were studied: the optimum reaction temperature was 45 鈩，
本文編號：2316294
[Abstract]:The degradation of pollutants by white rot fungi has the characteristics of non-specificity, high efficiency and broad spectrum, so it has a good application prospect in the bioremediation of chlorophenol refractory organic compounds. White rot fungi and laccase have great research value and application potential in industrial wastewater treatment and pulp biological bleaching. However, laccase production is generally low in production process, high production cost and other problems. In this paper, by adding cheap natural substrates and metal ions, the enzyme production medium was optimized to increase the laccase yield, and various factors were optimized, and the enzyme production law of white rot fungi was discussed based on the fermentation kinetics model. White rot fungi and laccase were used in chlorophenol wastewater respectively to study the bioremediation mechanism of white rot fungi and laccase. The aim is to provide some theoretical basis and technical support for the application of white rot fungi and laccase in wastewater treatment. In this paper, the fermentation conditions of laccase from Pleurotus strigella (Pleurotus ostreatus) were optimized by adding different natural substrates. The results showed that the optimum culture medium of orange peel was the best, the highest enzyme activity was increased from 5U/mL to 45 U / mL, and the fermentation time was shortened by 1 / 3. At the same time, the fermentation kinetics was studied, and the two groups of dynamic models were obtained. Compared with A (orange peel), B (control), two groups of fermentation kinetics models were added, after the addition of orange peel, The ability of cell proliferation and laccase production were greatly improved, and the growth rate and yield coefficient were also improved to some extent. The degradation mechanism of chlorophenol wastewater remediated by white rot fungi was studied. It was found that the adsorption ability of P.ostreatus for chlorophenol / phenol was in the order of 2N 46-TCP2-CP phenol PCP;. Different chlorophenol addition methods did not result in a significant change in biomass. The enzyme activities in the 2-CP and 2H4 + 6-TCP groups on the 5th day were 10 times and 5 times higher than those in the gradually added 2-CP and 2H4 + 6-TCP groups, respectively, and were concentrated on the 5th day after chlorophenol was added. The three chlorophenol removal rates were improved in different degrees, so the inhibition of chlorophenol on laccase activity and the removal rate of chlorophenol could be reduced by changing the method of chlorophenol addition. The effect of P.ostreatus on the removal of chlorophenol was studied. When the concentration of chlorophenol was in the range of 10~30mg/L, the degradation of 2-CP accounted for the vast majority and the proportion of adsorption was small, and the proportion of adsorption and degradation was the same in the process of TCP degradation. Compared with adsorption, PCP degradation accounted for most of the proportion, and the degradation ratio decreased with the increase of concentration. The properties of crude enzyme were studied: the optimum reaction temperature was 45 鈩，
本文編號：2316294
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