壬基酚对一株铜绿假单胞菌吸附镉的影响

考察壬基酚（NP）对一株从重金属污染场地筛选出的铜绿假单胞菌X吸附镉（Cd）的影响,在NP浓度分别为0、1.0、10.0mg/L条件下,通过优化吸附条件,研究X菌的Cd²⁺吸附效果.结果表明,NP与Cd²⁺共存时,在Cd²⁺初始浓度为1.0mg/L的溶液中,菌体的最佳投菌量和pH值为1.0g/L和7.0,吸附2h,吸附率可达90.0%左右.失活菌与活菌的吸附结果表明失活菌的吸附能力较活菌强.NP浓度对吸附的影响结果表明,低浓度NP对菌株吸附Cd²⁺的抑制作用较小,高浓度NP时抑制作用较大.通过分析X菌处理单一Cd²⁺及Cd²⁺-NP复合污染后的红外光谱图可知,菌体表面的羟基O-H键、酰胺C-N键和N-H键和氨基均参与吸附反应,并且高浓度NP对菌体表面基团活性影响较大,从而影响其对Cd²⁺的吸附.     

Pseudomonas putida细胞对微囊藻毒素-LR的胁迫响应

将1.0g/L的微囊藻毒素-LR(MC-LR)降解菌恶臭假单胞菌(Pseudomonas putida)置于含不同浓度MC-LR的体系中,研究了体系中菌体细胞完整性和生物量的变化,考察了MC-LR对细胞的氧化胁迫以及抗氧化酶的响应.结果表明,MC-LR能够增大P. putida细胞质膜通透性,造成膜损伤,导致胞内物质外流,使细胞完整性遭到破坏;同时,MC-LR能够引起P. putida细胞的氧化胁迫,随着毒素暴露时间的延长,活性氧自由基(ROS)和膜脂过氧化产物丙二醛(MDA)含量显著升高,具有明显的剂量效应.超氧化物歧化酶(SOD)活性在MC-LR的诱导下有一个先升后降的过程,表现为对低浓度污染物的主动响应,而高浓度(2.5 mg/L)MC-LR作用5d后,ROS积累到相当高水平,对细胞代谢功能造成破坏,使SOD活性下降,并加速细胞的死亡,P. putida生物量与对照相比,下降了将近50%.     

荧光假单胞菌高效广谱载体的构建及分析

为促进高GC含量基因在荧光假单胞菌(Pseudomonas fluorescens)中表达效果更加理想、操作更加简便,本研究首先采用不依赖基因序列和连接反应的克隆(Sequence and ligation independent cloning,SLIC)方法将载体pCIBhis上与复制相关的序列和标记基因片段构建成克隆载体pCIBS1.然后优化荧光假单胞菌转化方法,用电转化法将pCIBS1导入荧光假单胞菌BL915中,随后又将T7和tac基因启动子分别插入pCIBS1中,成功构建了表达载体pCIBS3和pCIBS2.研究发现载体pCIBS1在大肠杆菌和荧光假单胞菌中均较为稳定,并且将绿色荧光蛋白基因插入表达载体中,在大肠杆菌BL21(DE3)中获得表达,验证了表达载体功能.本研究构建的表达载体和建立的荧光假单胞菌BL915电转化方法,为高GC含量基因在荧光假单胞菌中的表达奠定了基础.     

一株苯乙烯高效降解菌的分离鉴定及降解特性研究

从某橡胶厂的活性污泥中筛选得到一株可高效降解苯乙烯的微生物菌株WJ,通过透射电镜(TEM)、生理生化试验、Biolog鉴定和16S rRNA基因的系统发育树建立等分析方法,鉴定该菌株为恶臭假单胞菌(Pseudomonas putida).通过环境单因素试验,确定该菌株WJ在温度30℃、pH值7.0的培养条件下具有良好的降解效果.通过不同初始浓度苯乙烯的降解及生长特性,确定该菌株的最大底物耐受浓度达到1500mg/L,且其生长动力学过程符合Haldane’s方程,相应动力学参数为: vmax=0.282h-1,Ks=23.57mg/L,Ki=1784.56mg/L.底物宽泛性试验表明,该菌株对苯系物(BTEX)、正己烯等与苯乙烯结构类似的碳氢化合物都具有良好的降解能力,表明其具备良好的底物宽泛性.该菌株对苯乙烯的氧化降解途径主要为乙烯基侧链的氧化.     

Toxic effect of sodium perfluorononyloxy-benzenesulfonate on Pseudomonas stutzeri in aerobic denitrification,cell structure and gene expression

• OBS inhibited the growth of P. stutzeri and destroyed its structure. • OBS was toxic to the aerobic denitrification process of P. stutzeri. • OBS induced the production of ROS in P. stutzeri. • OBS affected the expression of key genes involved in denitrification and SOD. The toxicities of sodium perfluorononyloxy-benzenesulfonate (OBS) to animals and plants are similar to those of perfluorooctane sulfonate. However, the mechanism of its toxicity to aerobic denitrifying bacteria is still unclear. In the present study, the ecotoxicity of OBS on an aerobic denitrifying strain, Pseudomonas stutzeri, was evaluated. The results showed that a dosage of OBS clearly affected the growth and aerobic denitrification of P. stutzeri. When compared with an unamended control, the degradation efficiency of the total nitrogen decreased by 30.13% during exposure to 200 mg/L of OBS, and the complete degradation time of nitrate-nitrogen was delayed by 4 h. The lactate dehydrogenase and superoxide dismutase produced by the bacteria increased with the concentration of OBS, and reactive oxygen species were also detected by confocal laser scanning microscope imaging. Transmission electron microscope imaging revealed chromosome deformation of the cells and damage to cell content; moreover, outer membrane vesicles were secreted from the bacteria, which was the important detoxification mechanism of P. stutzeri to OBS. Expression of the genes involved in aerobic nitrification and oxidative stress were also changed under OBS stress, which further confirmed the toxicity of OBS to P. stutzeri. This study reveals the environmental exposure risk of OBS from the perspective of microorganisms.     

Effects of microbial inoculations and surfactant levels on biologically- and chemically-assisted phytoremediation of lead-contaminated soil by maize (Zea Mays L.)

Assisted phytoremediation has been widely used for decontamination of potentially toxic elements contaminated soils. A greenhouse experiment was conducted to evaluate the effectiveness of different microbial inoculations and surfactant levels on the phytoremediation of a Pb-polluted calcareous soil by maize. The results showed that application of surfactant increased both root and shoot dry matter yields. Microbial inoculations, however, had no significant effect on the root or shoot dry matter yield. Mean Pb uptake in maize root or shoot increased only following the application of some surfactant levels. Inoculation with microorganisms significantly increased both mean Pb concentration and uptake in maize root but not in maize shoot. Application of 4?mmol surfactant kg?1 along with inoculation with Priformospora indica was effective in increasing Pb phytostabilisation potential. While the application of 2?mmol surfactant kg^?1 along with inoculation with Pseudomonas fluorescens was effective in increasing Pb phytoextraction potential. The fact that the values of translocation efficiencies were low in all treatments, demonstrated the low capability of maize for translocation of Pb from root to shoot. Inoculation with P. fluorescens was the most effective treatment in increasing metal micronutrient uptake. Microbial inoculation and surfactant levels enhanced Pb phytoremediation mostly through phytostabilisation of this metal by maize.     

两株假单胞菌降解酚类化合物的特性

从焦化废水中分离得到2株可降解对氯酚的假单胞菌(Pseudomonas)CO-1和CO-44.其最适降解温度为35~40℃,最适pH值为6.0~8.0.菌株降解对氯酚的速度与对氯酚初始浓度呈负相关.2株细菌均能较快地降解苯酚和甲酚,其中CO-1还能够降解1-萘酚和萘.在添加对氯酚的焦化废水中,CO-1和CO-44能够在42h内将苯酚、甲酚和对氯酚完全降解.从2株细菌中均检测到了苯酚羟化酶基因,分别从菌株CO-1和CO-44中检测到邻苯二酚1,2-双加氧酶基因和邻苯二酚2,3-双加氧酶基因.     

Experimental study on Cr(Ⅵ) reduction by Pseudomonas aeruginosa

Investigation on Cr(Ⅵ) reduction was conducted using Pseudomonas aeruginosa. The study demonstrated that the Cr(Ⅵ) can be effectively reduced to Cr(Ⅲ) by Pseudomonas aeruginosa. The effects of the factors affecting Cr(Ⅵ) reduction rate including carbon source type, pH, initial Cr(Ⅵ) concentration and amount of cells inoculum were thoroughly studied. Malate was found to yield maximum biotransformation, followed by succinate and glucose, with the reduction rate of 60.86%, 43.76% and 28.86% respectively. The optimum pH for Cr(Ⅵ) reduction was 7.0, with reduction efficiency of 61.71% being achieved. With the increase of initial Cr(Ⅵ) concentration, the rate of Cr(Ⅵ) reduction decreased. The reduction was inhibited strongly when the initial Cr(Ⅵ) concentration increased to 157 mg/L. As the amount of cells inoculum increased, the rate of Cr(Ⅵ) reduction also increased. The mechanism of Cr(Ⅵ) reduction and final products were also analysed. The results suggested that the soluble enzymes appear to be responsible for Cr(Ⅵ) reduction by Pseudomonas aeruginosa, and the reduced Cr(Ⅲ) was not precipitated in the form of Cr(OH)3.     

废纸造纸循环废水高效好氧菌的筛选和研究

利用平板分离方法,从废纸造纸封闭循环废水处理的生物接触氧化池中筛选出优势的高效菌株,采用形态学观察和生理生化试验发现主要由动胶菌属(Zoogloea sp.)和假单胞菌属(Pseudomona sp.)组成。用吸光光度法,确定其最佳生长时间。通过平行对比实验,在不同时间取样测定COD值,比较得出高效菌对废纸造纸循环废水的处理能力远高于普通菌。     

Biodegradation of anthracene and phenanthrene by bacteria isolated from oil-contaminated soil of Bangladesh

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous environmental pollutants entering into the environment through natural and anthropogenic activities. Owing to their toxicity to various life forms including humans, detoxification of PAHs is crucial to reduce their effects on cells. In this study, we have isolated two bacteria capable of degrading two common PAHs, anthracene and phenanthrene, from contaminated soil samples by using selective enrichment culture supplemented with test PAHs as the sole source of carbon and energy. The isolated bacteria were identified and affiliated as Pseudomonas aeruginosa strain KD and Stenotrophomonas maltophilia strain RC based on their 16s rRNA gene sequences. The degradation of anthracene and phenanthrene was estimated indirectly by measuring the decolourisation extent of a redox indicator, 2, 6-dichlorophenolindophenol, incorporated into PAH-supplemented mineral salt media. In the case of anthracene, ≥90% decolourisation was recorded at 20 and 48 days for P. aeruginosa and S. Maltophilia, respectively. On the other hand, ≥94% decolourisation was recorded at 56 and 52 days for P. aeruginosa and S. maltophilia, respectively during the utilisation of phenanthrene.