Biotransformation of phenylurea herbicides by a soil bacterial strain, Arthrobacter sp. N2: structure, ecotoxicity and fate of diuron metabolite with soil fungi

In order to assess the influence of the aromatic substitution on the ability of a soil bacterial strain, Arthrobacter sp. N2, to degrade phenylurea herbicides, biotransformation assays were performed in mineral medium with resting cells of this soil bacterial strain on three phenylurea herbicides (diuron, chlorotoluron and isoproturon). Each herbicide considered, led to the formation of only one metabolite detected by HPLC analysis. After isolation, the metabolites were identified by NMR and MS, as the corresponding substituted anilines. According to the Microtox test (realized on the bacterium Vibrio fischeri), these metabolites presented non-target toxicity far more important (up to 600 times higher for 4-isopropylaniline) than the parent molecule. For isoproturon and chlorotoluron, the amount of substituted anilines obtained at the end of the biotransformation was very low, whereas the biotransformation of diuron into 3,4-dichloroaniline was almost quantitative. In this last case, the degradation product accumulated in the medium. In soil, other microorganisms are present that might degrade it. So the biotransformation of 3,4-dichloroaniline was then tested with four fungal strains: Aspergillus niger, Beauveria bassiana, Cunninghamella echinulata var. elegans and Mortierella isabellina. The aniline was further transformed with all the microorganisms tested. Only one metabolite was detected by HPLC analysis and after isolation, it was identified to be 3,4-dichloroacetanilide. This acetylated compound led to biological effects less important on V. fischeri than 3,4-dichloroaniline. These results stress the importance of identifying the degradation products to assess the impact of a polluting agent. Indeed, the pollutant may undergo transformation yielding compounds more toxic than the parent molecule.     

一株1，2-二氯苯降解菌的分离鉴定及其降解特性

采用富集驯化方法,从盐城芦苇湿地根际土壤中分离得到一株可高效降解1,2-二氯苯的菌株,命名为DL-1。该菌株可以在以1,2-二氯苯为惟一碳源的无机培养基上生长,能够耐受最高浓度为200 mg/L的1,2-二氯苯。根据形态特征观察、生理生化鉴定和16S rDNA序列同源性分析,该目标菌株被鉴定为蜡质芽孢杆菌（Bacillus cereus）。菌株DL-1对1,2-二氯苯降解性能研究表明,该菌株为一株兼性厌氧菌,其适宜降解浓度、适宜温度、适宜pH值和适宜接种量分别为120mg/L、32℃、7和10%,在适宜降解条件下降解12,-二氯苯4 d其降解率达到80.3%。本实验为利用该菌株降解12,-二氯苯污水的应用提供了理论基础。     

对氨基苯磺酸降解菌的分离及其特性研究

从温州地区受污染的河水中分离到一株能降解对氨基苯磺酸的菌株WZR-3,该菌株能以对氨基苯磺酸为惟一碳源、能源生长。经对其形态特征、生理生化以及16S rDNA序列分析,该菌株初步鉴定为人苍白杆菌(Ochrobactrum anthropi)。该菌株利用对氨基苯磺酸生长时最适生长温度和pH值分别为30℃和7。该菌在10 g/L对氨基苯磺酸时仍能生长,最适生长浓度为300 mg/L对氨基苯磺酸。降解底物广谱性测试表明,该菌株还能降解多种芳香类化合物。     

Studies on biodegradation of nicotine by Arthrobacter sp. strain HF-2

A nicotine-degrading bacterium, strain HF-2, was isolated from tobacco waste-contaminated soil and identified as a member of Arthrobacter sp. based on morphology, physiological tests, 16S rDNA sequence and phylogenetic characteristics. At thermal denaturation test indicated that the G + C mol% of strain HF-1 was 63.5. The relationship between the growth of the isolate and the nicotine degradation suggested that strain HF-2 could utilize nicotine as sole sources of carbon, nitrogen and energy. Blue pigment was observed during the nicotine degradation by strain HF-2. The isolate grew well at 20 to 33 degrees C, initial pH 6.5 to 8.0 and 0.5 to 2.0 g L-1 of nicotine concentration in the nicotine inorganic salt media. The maximum growth and nicotine degradation occurred at 30 degrees C, initial pH 7.0 and 0.7 g.L-1 of nicotine concentration in media under natural incubation condition. Strain HF-2 could degrade 100% of nicotine under the optimized incubation conditions for 43 h. The concentrations of nicotine were monitored by high performance liquid chromatography. This study demonstrates Arthrobacter sp. strain HF-2 had a great ability to degrade nicotine, and it may be available for the application to the bioremediation of environments contaminated by tobacco waste.     

一株苯胺降解菌的分离鉴定及其降解特性

为研究苯胺污染的生物控制,通过驯化培养,从南京化工厂污水处理厂的活性污泥中分离出一株高效苯胺降解菌——菌株AN4.生理生化试验鉴定菌株AN4为金黄杆菌属(Chryseobacterium sp.).菌株AN4利用苯胺生长和降解苯胺的最适温度为30 ℃、pH为7.0,它可在苯胺质量浓度低于3 000 mg/L的无机盐固体培养基上生长.菌株AN4除可降解苯胺外,还可以苯酚、苯甲酸、硝基苯、甲苯、萘、氯苯、二甲苯作为唯一碳源生长,蛋白胨可加速其对苯胺的代谢.代谢机制研究证实,菌株AN4在邻苯二酚1,2-双加氧酶作用下经邻位裂解途径降解苯胺.     

一株生物表面活性剂产生菌的筛选鉴定及其特性

利用蓝色凝胶平板筛选法,从华北某油田受污染的土壤中分离筛选得到1株优良的生物表面活性剂产生菌H1。通过生理生化特征及16S rRNA基因序列相似性分析,将其鉴定为肺炎克雷伯氏菌(Klebsiella pneumoniae)。薄层层析和红外光谱分析表明,该菌株所产生物表面活性剂为磷脂类生物表面活性剂。对其所产生物表面活性剂的稳定性进行研究,并考察影响该生物表面活性剂合成的因素。稳定性实验显示,该生物表面活性剂可耐受90℃的高温,对pH有较广泛的适应性(pH 6.5~11.0),NaCl浓度对其生物活性影响不大。以蔗糖为碳源,硝酸铵为氮源,初始pH 6.5~7.0,30℃的培养条件有利于该生物表面活性剂的合成,生物表面活性剂的产量可达0.742 g/L。该研究成果可为原油的驱油降黏提供有效的菌源和理论依据。     

一株具有荧蒽降解能力的产吲哚乙酸菌的筛选鉴定及其特性

从植物根际土壤中筛选出能分泌吲哚乙酸(3-indole acetic acid,IAA),并对荧蒽具有一定降解效果的菌株ZZ21.30℃、180 r/min摇床培养24 h,能产生39.81 mg/L IAA,静息细胞对50 mg/L荧蒽转化7 d后,降解率达25.61%.此外,该菌还耐盐碱,能解磷,在pH为10、含盐度为7%的LB培养基上能够生长,30℃培养72h对磷酸三钙液体培养基的溶磷量达120.86 mg/L.经形态学观察、部分生理生化特征测定以及16S rDNA的保守序列鉴定,初步确定该菌株为耐盐节杆菌(Arthrobacter pascens).为促进菌株生长,提高菌株IAA产量,设定了基于培养时间、初始pH、通气量、温度、不同碳源和氮源的单因素菌株发酵条件优化实验,结果表明,促进菌株生长和提高IAA分泌的最佳培养条件并不完全一致,综合考量得到最佳发酵培养条件是初始pH为6,装液量为45 mL/150 mL,28℃摇床培养15 h,最佳碳、氮源分别是甘露醇和酵母粉.在最佳单因素优化条件下菌株产IAA能力最强可达92.31 mg/L.     

一株荧蒽降解菌的筛选鉴定及降解特性

采用富集培养方法,从多环芳烃污染的土壤中分离筛选到一株能以高分子量的多环芳烃荧蒽为惟一碳源和能源且生长状况良好的菌株DN002。通过形态观察、生理生化指标及16S rRNA同源序列分析比对,结果表明,该菌株为木糖氧化产碱菌(Achromobacter xylosoxidans),最适生长温度为32℃,最适生长pH为7～7.5。该菌株对荧蒽有较强的降解能力,在14 d内对500 mg/L的荧蒽的降解率为92.8%。菌株细胞蛋白SDS-PAGE结果显示,经荧蒽诱导后,在分子量为18～66 kDa范围内有显著的差异蛋白条带。     

一株微囊藻毒素降解菌的分离与鉴定

以水华蓝藻细胞中提取的微囊藻毒素为筛选物质,从太湖水华腐烂蓝藻中富集筛选出1株微囊藻毒素降解菌。该菌株革兰氏染色呈阴性,细胞细长杆状,菌落黄色,圆形,不透明,接触酶、氧化酶实验均呈阳性。16S rRNA基因序列的长度为1 416 bp(GenBank登录号为FJ976656)。系统发育树显示,该菌株与微嗜酸寡养单胞菌(Stenotrophomonas acidam-iniphila)的亲缘关系最近。通过菌株形态特征、生理生化特征和16S rRNA基因序列分析,将此菌株鉴定为微嗜酸寡养单胞菌,不同于已报道的微囊藻毒素降解菌属种。微囊藻毒素降解实验表明,该菌株5 d内将15.4 mg/L的微囊藻毒素完全降解,降解能力高于假单胞菌。     

Environmental impact of diuron transformation: a review

Diuron is a biologically active pollutant present in soil, water and sediments. A synthesis of literature data on its physicochemical properties, partitioning behaviour, abiotic and biotic transformations, toxicological and ecotoxicological impacts has been here performed. Data have shown that diuron is generally persistent in soil, water and groundwater. It is also slightly toxic to mammals and birds as well as moderately toxic to aquatic invertebrates. However, its principal product of biodegradation, 3,4-dichloroaniline exhibits a higher toxicity and is also persistent in soil, water and groundwater. Thus, diuron indirectly possesses a significant amount of toxicity and could be a potential poisoning pesticide contaminant of groundwater. Unfortunately, groundwater contamination will still persist despite the progressive suppression of diuron (Directive 200/60/CE). Therefore, determining the main factors influencing its degradation and its ecotoxicological effects on the environment and health could provide a basis for further development of bioremediation processes.     

Isolation and characterization of an Alcaligenes sp. strain DG-5 capable of degrading DDTs under aerobic conditions

In this study, an Alcaligenes sp. strain DG-5 that can effectively degrade dichlorodiphenyltrichloro-ethanes (DDTs) under aerobic conditions was isolated from DDTs-contaminated sediment. Various factors that affect the biodegradation of DDTs by DG-5 were investigated. About 88 %, 65 % and 45 % of the total DDTs were consumed within 120 h when their initial concentrations were 0.5, 5 and 15 mg L?1, respectively. However, almost no degradation was observed when their concentration was increased to 30 mg L?1, but the addition of nutrients significantly improved the degradation, and 66 % and 90 % of the total DDTs were degraded at 336 h in the presence of 5 g L?1 peptone and yeast extract, respectively. Moreover, the addition of 20 mM formate also enhanced the ability of DG-5 to transform DDTs, and its DDT transformation capacity (T(c)) value was increased by 1.8 - 2.7 fold for the pure (p,p'-DDT or o,p'-DDT only) and mixed systems (p,p'-DDT, o,p'-DDT, p,p'-DDD and p,p'-DDE). Furthermore, it was found that competitive inhibition in the biodegradation by DDT compounds occurred in the mixed system.     

Isolation and characterization of an Alcaligenes sp. strain DG-5 capable of degrading DDTs under aerobic conditions

In this study, an Alcaligenes sp. strain DG-5 that can effectively degrade dichlorodiphenyltrichloro-ethanes (DDTs) under aerobic conditions was isolated from DDTs-contaminated sediment. Various factors that affect the biodegradation of DDTs by DG-5 were investigated. About 88 %, 65 % and 45 % of the total DDTs were consumed within 120 h when their initial concentrations were 0.5, 5 and 15 mg L^?1, respectively. However, almost no degradation was observed when their concentration was increased to 30 mg L^?1, but the addition of nutrients significantly improved the degradation, and 66 % and 90 % of the total DDTs were degraded at 336 h in the presence of 5 g L^?1 peptone and yeast extract, respectively. Moreover, the addition of 20 mM formate also enhanced the ability of DG-5 to transform DDTs, and its DDT transformation capacity (T_c) value was increased by 1.8 - 2.7 fold for the pure (p,p’-DDT or o,p’-DDT only) and mixed systems (p,p’-DDT, o,p’-DDT, p,p’-DDD and p,p’-DDE). Furthermore, it was found that competitive inhibition in the biodegradation by DDT compounds occurred in the mixed system.     

一株具有藻毒素降解和溶藻功能细菌的分离鉴定

从太湖水华腐烂蓝藻中富集筛选出一株编号A-8的细菌。该菌株菌落圆形、灰白色,革兰氏染色阴性,氧化酶阴性,接触酶阳性;分子系统进化树显示为溶血不动杆菌。根据以上结果,将此菌株鉴定为溶血不动杆菌。此外,对这株菌降解微囊藻毒素、不同生长时期的细菌的溶藻效果及溶藻专一性进行了研究。结果显示:该菌在3 d时间里将藻毒素降解了30.4%;细菌通过分泌胞外物质溶藻,细菌在衰退期时胞外分泌物的溶藻效果最好;该菌还对瘤状念珠藻、颤藻等9种蓝藻有溶藻作用。     

1，4-二氯苯降解菌的分离及其降解特性研究

从某污水处理曝气池的活性污泥中分离出一株能够以1,4-二氯苯为唯一碳源和能源生长的菌株DEB-1,通过形态特征和生理生化试验初步鉴定为黄杆菌属（Flavobacterium sp.）。实验结果表明,该菌株最适降解温度为32℃、最适降解pH为7.8,24 h对100 mg/L的1,4-二氯苯的降解率达94.5%。菌株DEB-1的降解谱较广,对5种氯苯类物质具有较高的降解率。并进一步研究了DEB-1的1,4-二氯苯降解酶粗酶液的性质,其最适反应温度和pH分别为30℃和8.5。对处理含氯代芳香化合物的有机废水具有一定的意义。     

Studies on Biodegradation of Nicotine by Arthrobacter sp. Strain HF-2

A nicotine-degrading bacterium, strain HF-2, was isolated from tobacco waste-contaminated soil and identified as a member of Arthrobacter sp. based on morphology, physiological tests, 16S rDNA sequence and phylogenetic characteristics. At thermal denaturation test indicated that the G + C mol% of strain HF-1 was 63.5. The relationship between the growth of the isolate and the nicotine degradation suggested that strain HF-2 could utilize nicotine as sole sources of carbon, nitrogen and energy. Blue pigment was observed during the nicotine degradation by strain HF-2. The isolate grew well at 20 to 33°C, initial pH 6.5 to 8.0 and 0.5 to 2.0 g L^?1 of nicotine concentration in the nicotine inorganic salt media. The maximum growth and nicotine degradation occurred at 30°C, initial pH 7.0 and 0.7 g·L^?1 of nicotine concentration in media under natural incubation condition. Strain HF-2 could degrade 100% of nicotine under the optimized incubation conditions for 43 h. The concentrations of nicotine were monitored by high performance liquid chromatography. This study demonstrates Arthrobacter sp. strain HF-2 had a great ability to degrade nicotine, and it may be available for the application to the bioremediation of environments contaminated by tobacco waste.     

Nitrogen impacts on atrazine-degrading Arthrobacter strain and bacterial community structure in soil microcosms

The objective of this study was to investigate the impacts of exogenous nitrogen on a microbial community inoculated with the atrazine-degrading Arthrobacter sp. in soil amended with a high concentration of atrazine. Inoculated and uninoculated microcosms for biodegradation tests were constructed. Atrazine degradation capacity of the strain DAT1 and the strain's atrazine-metabolic potential and survival were assessed. The relative abundance of the strain DAT1 and the bacterial community structure in soils were characterized using quantitative PCR in combination with terminal restriction fragment length polymorphism. Atrazine degradation by the strain DAT1 and the strain's atrazine-metabolic potential and survival were not affected by addition of a medium level of nitrate, but these processes were inhibited by addition of a high level of nitrate. Microbial community structure changed in both inoculated and uninoculated microcosms, dependent on the level of added nitrate. Bioaugmentation with the strain DAT1 could be a very efficient biotechnology for bioremediation of soils with high concentrations of atrazine.     

卡马西平降解菌的筛选分离及其降解机理

药品污染物日益成为新兴污染物研究的重点,药品卡马西平因具有多种药效被广泛使用,在环境中频繁被检出,且浓度较高,不易去除,通常作为环境中药品污染状况的指示化合物。本研究从长期用于去除药品废水的曝气生物滤池中分离出一株细菌YK-6,其能以卡马西平为惟一碳源、氮源和能源生长,通过生理生化以及16S rDNA基因序列分析鉴定并命名为Pseudomonas putida YK-6。该菌株YK-6在pH为7.2、温度30℃、卡马西平初始浓度为20 mg/L、摇床振荡速率为160 r/min的生长条件下培养5 d,对卡马西平的降解率可达54.66%。菌株YK-6对卡马西平的可能降解途径为首先通过生物的氧化作用,将CBZ氧化成CBZ-EP,CBZ-EP经过水解作用转化为CBZ-DiOH,CBZ-DiOH经丙酮酸氧化脱羧及在NADH还原性辅酶的作用下,裂解成苯胺和邻苯甲酸,苯胺和邻苯甲酸再经进一步氧化,直至最终矿化。