1株壬基酚降解菌的分离筛选鉴定及其降解特性研究
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摘要
目前,很多水体受到了内分泌干扰物(EDCs)的污染,由于其巨大的危害性,引起了国内外强烈的关注。
壬基酚(NP)是有代表性的环境内分泌干扰物和持久性有毒污染物,具有明显的雌激素效应和很强的生物积累性。它主要来源于广泛使用的非离子表面活性剂壬基酚聚氧乙烯醚的降解产物,每年都有大量的壬基酚聚氧乙烯醚排放到水体中,国内外很多的水体因此受到了壬基酚的污染。
从长期稳定运行并用三种不同方法处理(松花江原水、臭氧预氧化和高铁酸钾预氧化)的饮用水处理系统颗粒活性炭上分别分离出4株、13株和17株菌,高铁酸钾预氧化处理的生物活性炭上的生物量最大和生物相最复杂,常规处理的生物活性炭上生物量最少,生物相最简单。
利用自配的矿质盐培养基溶液(MSM)并以NP为唯一碳源对分离出的34株菌进行了筛选,得到一株NP高效降解菌NP-1,该菌来源于臭氧预氧化处理的活性炭。在NP初始浓度1.0mg/L,初始pH值6.0,培养温度30℃和摇床转速140r/min条件下,2%的菌量10d之内能够降解59%的NP,采用动力学模型对数据进行拟合,发现菌的降解过程基本满足一级动力学模型,其降解速率常数(k)为0.0865d~(-1),半衰期(t_(1/2))为8.0d。
菌NP-1是一株革兰氏阳性菌,显微镜下观察为短杆菌,无鞭毛,尺寸大约:1.77×0.64μm。利用Sherlock MIS系统鉴定菌NP-1为红串红球菌。
通过摇瓶实验考察各影响因素,得出最佳降解条件是温度30℃,pH 6.0。菌NP-1降解NP的速率与NP的初始浓度关系不大但随着初始菌量的增加而显著提高。MSM溶液中的NH_4~+、Mn~(2+)、Mg~(2+)、Na+离子和葡萄糖、醋酸钠和酵母膏等底物对菌降解NP均有促进作用;而Ca~(2+)、Cu~(2+)、Fe~(2+)和磷酸盐的作用则表现为抑制。混合菌体系能够更好的适应环境并拥有更好的降解性能,降解NP速率与体系中含有菌NP-1的绝对数量有关。
本研究首次从给水处理系统长期运行的生物活性炭上分离得到1株NP高效降解菌,并对其进行初步的研究,以期揭示NP生物降解的基本规律,并为其在生物修复和水中NP去除的应用中提供一定的参考。
At present, a number of rivers are polluted by Endocrine Disrupting Chemicals (EDCs). Because of their high toxicity, more and more attention has been paid to the problem.
Nonylphenol (NP) is one of the representative endocrine disrupting chemicals and persistent toxic pollutants, with obvious estrogenic activity and high bioaccumulation in body. Nonylphenol in water is mainly genereated by the degradation of nonylphenol polyethoxylates, which is widely used as nonionic surfactant in various industries. It was reported that large amount of such surfactant was let into nature, and more and more rivers were polluted by NP, and it has become a big international problem.
There were 4, 13 and 17 bacterial strains isolated from activated carbon, which was processed with sand filtered water from Songhua River for long time, pre-oxidized with ozone and ferrate respectively, and it demonstrated that treatment effect of water pre-oxidized with ferrate was better than that of pre-oxidized with ozone.
A bacterial strain NP-1, capable of utilizing nonylphenol (NP) as solo carbon source, was isolated using mimeral salt medium (MSM) by adding NP as solo carbon source, and the bacterial strain was from the ozone-activated carbon column. Under conditions of pH 6.0, 30℃, 140r/min, and 2% of inoculum amount, the strain could biodegrade 59% of 1.0 mg/L NP, and the biodegradation reaction fitted well with first-order kinetic model, with a degradation rate constant (k) of 0.0865d~-1) and a half-live (t_(1/2)) of 8.0d.
The bacterial strain was gram-positive, and was bacillus without flagella, and the size was 1.77×0.64μm. The bacterial strain was identified as Rhodococcus-erythropolis with the Sherlock Microbial Identification System (MIS) from the MIDI Corporation.
Many factors were studied in shaking flasks, and the optimal conditions for NP biodegradation were at 30℃and pH 6.0. Aerobic degradation rate was inert to the original NP concentration, but was highly enhanced by increased inoculum amount. The aerobic degradation rate for NP was also enhanced by increased level of NH_4~+, Mn~(2+), Mg~(2+), and NaCl, and by addition of glucose, NaAc, and yeast extract, as well as by reduced level of Ca~(2+), Cu~(2+), Fe~(2+), and phosphate. The combination of NP- with other bacterial strains which were also isolated from activated carbon expressed better biodegradation effectiveness and could adapt to environment more easily.
This study isolated a NP biodegradation bacterial strain from activated carbon in the long-running drinking water treatment system for the first time, and explored NP biodegradation principles, and provided some reference for NP removal in water.
壬基酚(NP)是有代表性的环境内分泌干扰物和持久性有毒污染物,具有明显的雌激素效应和很强的生物积累性。它主要来源于广泛使用的非离子表面活性剂壬基酚聚氧乙烯醚的降解产物,每年都有大量的壬基酚聚氧乙烯醚排放到水体中,国内外很多的水体因此受到了壬基酚的污染。
从长期稳定运行并用三种不同方法处理(松花江原水、臭氧预氧化和高铁酸钾预氧化)的饮用水处理系统颗粒活性炭上分别分离出4株、13株和17株菌,高铁酸钾预氧化处理的生物活性炭上的生物量最大和生物相最复杂,常规处理的生物活性炭上生物量最少,生物相最简单。
利用自配的矿质盐培养基溶液(MSM)并以NP为唯一碳源对分离出的34株菌进行了筛选,得到一株NP高效降解菌NP-1,该菌来源于臭氧预氧化处理的活性炭。在NP初始浓度1.0mg/L,初始pH值6.0,培养温度30℃和摇床转速140r/min条件下,2%的菌量10d之内能够降解59%的NP,采用动力学模型对数据进行拟合,发现菌的降解过程基本满足一级动力学模型,其降解速率常数(k)为0.0865d~(-1),半衰期(t_(1/2))为8.0d。
菌NP-1是一株革兰氏阳性菌,显微镜下观察为短杆菌,无鞭毛,尺寸大约:1.77×0.64μm。利用Sherlock MIS系统鉴定菌NP-1为红串红球菌。
通过摇瓶实验考察各影响因素,得出最佳降解条件是温度30℃,pH 6.0。菌NP-1降解NP的速率与NP的初始浓度关系不大但随着初始菌量的增加而显著提高。MSM溶液中的NH_4~+、Mn~(2+)、Mg~(2+)、Na+离子和葡萄糖、醋酸钠和酵母膏等底物对菌降解NP均有促进作用;而Ca~(2+)、Cu~(2+)、Fe~(2+)和磷酸盐的作用则表现为抑制。混合菌体系能够更好的适应环境并拥有更好的降解性能,降解NP速率与体系中含有菌NP-1的绝对数量有关。
本研究首次从给水处理系统长期运行的生物活性炭上分离得到1株NP高效降解菌,并对其进行初步的研究,以期揭示NP生物降解的基本规律,并为其在生物修复和水中NP去除的应用中提供一定的参考。
At present, a number of rivers are polluted by Endocrine Disrupting Chemicals (EDCs). Because of their high toxicity, more and more attention has been paid to the problem.
Nonylphenol (NP) is one of the representative endocrine disrupting chemicals and persistent toxic pollutants, with obvious estrogenic activity and high bioaccumulation in body. Nonylphenol in water is mainly genereated by the degradation of nonylphenol polyethoxylates, which is widely used as nonionic surfactant in various industries. It was reported that large amount of such surfactant was let into nature, and more and more rivers were polluted by NP, and it has become a big international problem.
There were 4, 13 and 17 bacterial strains isolated from activated carbon, which was processed with sand filtered water from Songhua River for long time, pre-oxidized with ozone and ferrate respectively, and it demonstrated that treatment effect of water pre-oxidized with ferrate was better than that of pre-oxidized with ozone.
A bacterial strain NP-1, capable of utilizing nonylphenol (NP) as solo carbon source, was isolated using mimeral salt medium (MSM) by adding NP as solo carbon source, and the bacterial strain was from the ozone-activated carbon column. Under conditions of pH 6.0, 30℃, 140r/min, and 2% of inoculum amount, the strain could biodegrade 59% of 1.0 mg/L NP, and the biodegradation reaction fitted well with first-order kinetic model, with a degradation rate constant (k) of 0.0865d~-1) and a half-live (t_(1/2)) of 8.0d.
The bacterial strain was gram-positive, and was bacillus without flagella, and the size was 1.77×0.64μm. The bacterial strain was identified as Rhodococcus-erythropolis with the Sherlock Microbial Identification System (MIS) from the MIDI Corporation.
Many factors were studied in shaking flasks, and the optimal conditions for NP biodegradation were at 30℃and pH 6.0. Aerobic degradation rate was inert to the original NP concentration, but was highly enhanced by increased inoculum amount. The aerobic degradation rate for NP was also enhanced by increased level of NH_4~+, Mn~(2+), Mg~(2+), and NaCl, and by addition of glucose, NaAc, and yeast extract, as well as by reduced level of Ca~(2+), Cu~(2+), Fe~(2+), and phosphate. The combination of NP- with other bacterial strains which were also isolated from activated carbon expressed better biodegradation effectiveness and could adapt to environment more easily.
This study isolated a NP biodegradation bacterial strain from activated carbon in the long-running drinking water treatment system for the first time, and explored NP biodegradation principles, and provided some reference for NP removal in water.
引文
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