3-PBA降解菌BA3的降解特性及基因工程菌构建

从石油污染土壤中分离到1株能以3-苯氧基苯甲酸(3-PBA)为唯一碳源生长的细菌,命名为BA3.根据其生理生化特征和16S rDNA序列相似性分析,将该菌鉴定为鞘脂菌属(Sphingobium sp.).该菌株在60 h内对100 mg·L-1的3-苯氧基苯甲酸的降解率达到99%.降解3-苯氧基苯甲酸的最适温度为30℃...     

一株苯酚降解菌的分离和鉴定

:从工业废水中分离到一株高效降解苯酚的菌株PHEA 2 ,它能够在以苯酚为单一碳源和能源的无机培养基上生长 ,其最高苯酚降解率为 394nmol/ (mg·min) .经Biolog细菌自动鉴定仪鉴定 ,该菌株为醋酸钙不动杆菌 (Acinetobactercalcoaceti cus) .采用PCR扩增获得的该菌 16SrDNA片段 ,核苷酸序列分析结果表明 ,该菌株的 16SrDNA的核苷酸序列与醋酸钙不动杆菌同源性为 98% .在细菌系统发育分类学上 ,PHEA 2归属变形细菌γ亚类、不动杆菌属、醋酸钙不动杆菌 .     

己烯雌酚降解菌株沙雷氏菌的分离鉴定及其降解特性

利用具有降解己烯雌酚(diethylstilbestrol,DES)特性的功能微生物来降解DES、有望实现环境中DES的有效去除,然而迄今关于降解DES的功能菌株及其降解特性的报道很少.本研究通过选择性富集培养,从某污水处理站活性污泥中分离获得1株具有DES降解特性的细菌(菌株S).经形态观察、生理生化试验及16S rDNA序列同源性分析将菌株S鉴定为沙雷氏菌属(Serratia sp.)细菌.菌株S好氧生长,30℃、150 r·min-1摇床培养7 d后,对无机盐培养基中DES(50 mg·L-1)的降解率达68.3%.通过摇瓶实验,优化了菌株S生长和降解DES的最适环境条件:温度30℃,底物浓度40～60 mg·L-1,pH 7.0,接种量5%,盐添加量0 g·L-1,装液量10 mL(100 mL三角瓶).     

对苯二甲酸降解菌的筛选鉴定及其降解特性的研究

从本公司生物曝气池中筛选分离到了5株能高效降解对苯二甲酸(PTA)的菌株,编号为T1,T3,T5,T6,T7.利用16S rDNA测序后进行序列比对分析结果表明其分别为不动杆菌(Acinetobacter sp),不动杆菌(Acinetobacter sp),松鼠葡萄球菌(Staphylococcus sciuri),松鼠葡萄球菌(Staphylococcus sciuri),恶臭假单胞菌(Pseudomonas putida).在30度,pH7.5,1％接种量的条件下,培养72 h,PTA的降解率可达97％以上.对实际的印染废水中的PTA也有较好的降解效果,培养48 h后降解率最高可达65.6％.其中T1,T3有一定的实际应用价值.     

一株辛基酚聚氧乙烯醚降解菌的筛选、鉴定及其降解

采用TX-100为唯一碳源的无机盐培养基进行筛选,经过富集、分离和纯化获得了一株能够耐高浓度TX-100且降解性能较好的降解菌,命名为H1.对细菌H1进行形态学观察、生理生化鉴定和16S rDNA (GenBank Accession No.KC505179)序列相似性分析,初步鉴定为醋酸钙不动杆菌属(Acinetobacter calcoaceticus).通过对菌株H1降解TX-100的产物分析,初步推断TX-100的降解模式为末端氧化,先从EO末端氧化,醚键断裂,形成短链的化合物,同时释放出乙醛酸.质粒检测和消除试验表明,控制菌H1降解TX-100的基因位于质粒上.     

一株高效原油降解不动杆菌的筛选及降解特性分析

为得到高效原油降解菌,从天津大港油田废弃钻井附近的油污染土壤中筛选出一株高效原油降解菌YQJ-1.经生理生化实验、16S rDNA基因序列及系统发育树对该菌株鉴定分析,采用单因素实验研究环境因素对该菌株生长和原油降解特性的影响,并对动力学和烷烃降解进行了探讨。结果表明:该菌株为醋酸钙不动杆菌(Acinetobacter calcoaceticus);在35℃,pH为8,接种量为10mL,氮源为酵母浸粉时降解效果最佳;且1～5 g/L的原油降解与一级动力学模型一致,原油浓度为5g/L时的降解率高达77. 58%,石油降解半衰期仅为4. 93d; GC-FID分析发现,YQJ-1对链长C_(11)—C_(25)的烷烃平均降解率达到87. 84%。该研究为未来高浓度石油污染土壤环境修复提供了良好的候选菌株。     

多菌灵降解菌NY97-1的鉴定及降解条件

分离筛选出一株能高效降解多菌灵的芽孢杆菌菌株NY97-1,经生理生化和序列同源性分析,将该菌株鉴定为短小芽孢杆菌Bacillus pumilus.该细菌降解多菌灵的最适pH值为6.0～10.0,最适温度为35.0～40.0℃.该菌在多菌灵浓度为10、30、50、100、300 mg·L-1的无机盐培养基中,30℃振荡培养24 h后,其对多菌灵的降解率分别为42.44％、48.97％、77.19％、78.66％和90.07％.添加少量有机氮源如酵母浸出粉、胰蛋白胨、酵母膏可促进菌株NY97-1对多菌灵的降解作用,添加少量无机氮源尿素会抑制菌株NY97-1对多菌灵的降解作用.     

微囊藻毒素降解菌S3的分子鉴定及其降解毒素的研究

对一株具有强降解微囊藻毒紊MC-LR能力的细菌S3进行了分子鉴定.测得该菌16S rDNA为1396bp,GenBank序列登录号为DQ836314.序列比对结果显示,该菌与类芽孢杆菌Paenibacillus validus的相似性达98%.微囊藻毒素降解实验结果表明,该菌能在以微囊藻毒素为唯一碳、氮源的培养基中生长,微囊藻毒紊在72h内减少78.3%,菌株S3的最适生长温度是30℃,最适生长pH值为7.0.     

焦化废水中苯酚降解菌筛选及其降解性能

焦化废水中苯酚类及其衍生物的降解率高低是焦化废水COD是否达标排放的关键.采用不同培养基和菌种驯化方法,从焦化废水厂活性污泥中分离筛选获得4株苯酚降解菌,经生理生化和16S rDNA分子鉴定,A1为球杆菌属Sphaerobacter,C1为鲍曼不动杆菌Acinetobacter baumannii;D2为睾丸酮丛毛单胞菌Comamonas testosterone;D3为Novosphingobiumnaphthalenivorans.4株降酚菌均具有较高的苯酚耐受力和降解效率,是生物法处理酚类污染废水优质的种质资源.菌株D2不仅对苯酚具较高耐受力达到2 000 mg.L-1、且在48 h内可将初始浓度为1 000 mg.L-1的苯酚完全降解.环境因子考察研究表明,pH为7.5～8.5,温度为30～40℃范围内,转速为150 r.min-1,是菌株D2的最优降解条件,本研究结果为构建高效处理焦化废水基因工程菌提供了微生物基础.     

杀螟硫磷降解菌FDS-1的分离鉴定及其降解特性

从长期受杀螟硫磷污染的土壤中分离到一株能以杀螟硫磷为唯一碳源生长的细菌 FDS-1,根据其生理生化分析和 16S rDNA(GenBank Accession No. AY550913)序列同源性分析,将该菌初步鉴定为伯克霍尔德氏菌属(Burkholderia sp.).该菌能在 14h 内完全降解100mg/L 的杀螟硫磷.该菌降解杀螟硫磷最适 pH 值为 7.0,最适温度为 30℃,菌株降解杀螟硫磷的速率和起始接种量呈正相关.酶的定域试验表明,该菌中有机磷水解酶为胞内酶.     

Toxic effects of acetochlor, methamidophos and their combination on nifH gene in soil

Toxic effects of two agrochemicals on nifH gene in agricultural black soil were investigated using denaturing gradient gel electrophoresis （DGGE） and sequencing approaches in a microcosm experiment. Changes of soil nifH gene diversity and composition were examined following the application of acetochlor, methamidophos and their combination. Acetochlor reduced the nifH gene diversity （both in gene richness and diversity index values） and caused changes in the nifH gene composition. The effects of acetochlor on nifH gene were strengthened as the concentration of acetochlor increased. Cluster analysis of DGGE banding patterns showed that nifH gene composition which had been affected by low concentration of acetochlor （50 mg/kg） recovered firstly. Methamidophos reduced nifH gene richness that except at 4 weeks. The medium concentration of methamidophos （150 mg/kg） caused the most apparent changes in nifH gene diversity at the first week while the high concentration of methamidophos （250 mg/kg） produced prominent effects on nifH gene diversity in the following weeks. Cluster analysis showed that minimal changes of nifH gene composition were found at 1 week and maximal changes at 4 weeks. Toxic effects of acetochlor and methamidophos combination on nifH gene were also apparent. Different nifH genes （bands） responded differently to the impact of agrochemicals： four individual bands were eliminated by the application of the agrochemicals, five bands became predominant by the stimulation of the agrochemicals, and four bands showed strong resistance to the influence of the agrochemicals. Fifteen prominent bands were partially sequenced, yielding 15 different nifH sequences, which were used for phylogenetic reconstructions. All sequences were affiliated with the alpha- and beta-proteobacteria, showing higher similarity to eight different diazotrophic genera.     

Arsenic uptake by arbuscular mycorrhizal maize （Zea mays L.） grown in an arsenic-contaminated soil with added phosphorus

The effects of arbuscular mycorrhizal (AM) fungus (Glomus mosseae) and phosphorus (P) addition (100 mg/kg soil) on arsenic (As) uptake by maize plants (Zea mays L.) from an As-contaminated soil were examined in a glasshouse experiment.Non-mycorrhizal and zero-P addition controls were included.Plant biomass and concentrations and uptake of As,P,and other nutrients,AM colonization,root lengths,and hyphal length densities were determined.The results indicated that addition of P significantly inhibited root colonization and development of extraradical mycelium.Root length and dry weight both increased markedly with mycorrhizal colonization under the zero-P treatments,but shoot and root biomass of AM plants was depressed by P application.AM fungal inoculation decreased shoot As concentrations when no P was added,and shoot and root As concentrations of AM plants increased 2.6 and 1.4 times with P addition,respectively.Shoot and root uptake of P,Mn,Cu,and Zn increased,but shoot Fe uptake decreased by 44.6%,with inoculation, when P was added.P addition reduced shoot P,Fe,Mn,Cu,and Zn uptake of AM plants,but increased root Fe and Mn uptake of the nonmycorrhizal ones.AM colonization therefore appeared to enhance plant tolerance to As in low P soil,and have some potential for the phytostabilization of As-contaminated soil,however,P application may introduce additional environmental risk by increasing soil As mobility.     

Degradation of metabolites accumulated of benzo[a]pyrene by coupling Penicillium Chrysogenum with KMnO4

Several main metabolites of benzo[a]pyrene （BaP） formed by Penicillium chrysogenum, Benzo[a]pyrene-1,6-quinone （BP 1,6- quinone）, trans-7,8-dihydroxy-7,8-dihydrobenzo[a]pyrene （BP 7,8-diol）, 3-hydroxybenzo[a]pyrene （3-OHBP）, were identified by high-performance liquid chromatography （HPLC）. The three metabolites were liable to be accumulated and were hardly further metabolized because of their toxicity to microorganisms. However, their further degradation was essential for the complete degradation of BaP. To enhance their degradation, two methods, degradation by coupling Penicillium chrysogenum with KMnO4 and degradation only by Penicillium chrysogenum, were compared; Meanwhile, the parameters of degradation in the superior method were optimized. The results showed that （1） the method of coupling Penicillium chrysogenum with KMnO4 was better and was the first method to be used in the degradation of BaP and its metabolites; （2） the metabolite, BP 1,6-quinone was the most liable to be accumulated in pure cultures; （3） the effect of degradation was the best when the concentration of KMnO4 in the cultures was 0.01% （w/v）, concentration of the three compounds was 5 mg/L and pH was 6.2. Based on the experimental results, a novel concept with regard to the bioremediation of BaP-contaminated environment was discussed, considering the influence on environmental toxicity of the accumulated metabolites.     

Impact of atrazine and nitrogen fertilizers on the sorption of chlorotoluron in soil and model sorbents

Sorption of chlorotoluron in ammonium sulfate, urea and atrazine multi-solutes system was investigated by batch experiments. The results showed application of nitrogen fertilizers to the soil could affect the behavior of chlorotoluron. At the same concentration of N, sorption of chlorotoluron decreased as the concentration of atrazine increased on the day 0 and 6 in soil, respectively. The sorption of chlorotoluron increased from 0 to 6 d when soils were preincubated with deionized water, ammonium sulfate and urea solution for 6 d. That indicated incubation time was one of the most important factors for the sorption of chlorotoluron in nitrogen fertilizers treatments. The individual sorption isotherms of chlorotoluron in rubbery polymer and silica were strictly linear in single solute system, but there were competition sorption between pesticides or between pesticides and nitrogen fertilizers. That indicated the sorption taken place by concurrent solid-phase dissolution mechanism and sorption on the interface of water-organic matter or water-mineral matter.     

Potential of plant polyphenol oxidases in the decolorization and removal of textile and non-textile dyes

In this study an effort has been made to use plant polyphenol oxidases; potato （Solanum tuberosum） and brinjal （Solanum melongena）, for the treatment of various important dyes used in textile and other industries. The ammonium sulphate fractionated enzyme preparations were used to treat a number of dyes under various experimental conditions. Majority of the treated dyes were maximally decolorized at pH 3.0. Some of the dyes were quickly decolorized whereas others were marginally decolorized. The initial first hour was sufficient for the maximum decolorization of dyes. The rate of decolorization was quite slow on long treatment of dyes. Enhancement in the dye decolorization was noticed on increasing the concentration of enzymes. The complex mixtures of dyes were treated with both preparations of polyphenol oxidases in the buffers of varying pH values. Potato polyphenol oxidase was significantly more effective in decolorizing the dyes to higher extent as compared to the enzyme obtained from brinjal polyphenol oxidase. Decolorization of dyes and their mixtures, followed by the formation of an insoluble precipitate, which could be easily removed simply by centrifugation.     

Removal of heavy metals from sewage sludge by low costing chemical method and recycling in agriculture

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Single and joint effects of pesticides and mercury on soil urease

The influence of two pesticides including chlorimuron-ethyl and furadan and mercury （Hg） on urease activity in 4 soils （meadow burozem and phaeozem） was investigated. The soils were exposed to various concentrations of the two pesticides and Hg individually and simultaneously. Results showed that there was a close relationship between urease activity and organic matter content in soil. Chlorimuron-ethyl and furadan could both activate urease in the 4 soils. The maximum increment of urease activity by chlorimuronethyl was up to 14%-18%. There was almost an equal increase （up to 13%-21%） in the urease activity by furadan. On the contrary, Hg markedly inhibited soil urease activity. A logarithmic equation was used to describe the relationship （P〈0.05） between the concentration of Hg and the activity of soil urease in the 4 tested soils. Semi-effect dose （ED50） values by the stress of Hg based on the inhibition of soil urease in the 4 soils were 88, 5.5, 24 and 20 mg/kg, respectively, according to the calculation of the corresponding equations. The interactive effect of chlorimuron-ethyl or furadan with metal Hg on soil urease was mainly synergic at the highest tested concentrations.     

Isolation and preliminary characterization of a 3-chlorobenzoate degrading bacteria

A study was conducted to compare the diversity of 2-, 3-, and 4-chlorobenzoate degraders in two pristine soils and one contaminated sewage sludge. These samples contained strikingly different populations of mono-chlorobenzoate degraders. Although fewer cultures were isolated in the uncontaminated soils than contaminated one, the ability of microbial populations to mineralize chlorobenzoate was widespread. The 3- and 4-chlorobenzoate degraders were more diverse than the 2-chlorobenzoate degraders. One of the strains isolated from the sewage sludge was obtained. Based on its phenotype, chemotaxonomic properties and 16S rRNA gene, the organism S-7 was classified as Rhodococcus erythropolis. The strain can grow at temperature from 4 to 37℃. It can utilize several （halo）aromatic compounds. Moreover, strain S-7 can grow and use 3-chlorobenzoate as sole carbon source in a temperatures range of 10-30℃ with stoichiometric release of chloride ions. The psychrotolerant ability was significant for bioremediation in low temperature regions. Catechol and chlorocatechol 1,2-dioxygenase activities were present in cell free extracts of the strain, but no （chloro）catechol 2,3- dioxygenase activities was detected. Spectral conversion assays with extracts from R. erythropolis S-7 showed accumulation of a compound with a similar UV spectrum as chloro-cis,cis-muconate from 3-chlorobenzoate. On the basis of these results, we proposed that S-7 degraded 3-chlorobenzoate through the modified ortho-cleave pathway.     

Organochlorine pesticides in soils under different land usage in the Taihu Lake region, China

A field study was conducted in the Taihu Lake region, China in 2004 to reveal the organochlorine pesticide concentrations in soils after the ban of these substances in the year 1983. Thirteen organochlorine pesticides (OCPs) were analyzed in soils from paddy field, tree land and fallow land. Total organochlorine pesticide residues were higher in agricultural soils than in uncultivated fallow land soils. Among all the pesticides, ΣDDX (DDD, DDE and DDT) had the highest concentration for all the soil samples, ranging from 3.10 ng/g to 166.55 ng/g with a mean value of 57.04 ng/g and followed by ΣHCH, ranging from 0.73 ng/g to 60.97 ng/g with a mean value of 24.06 ng/g. Dieldrin, endrin, HCB and α-endosulfan were also found in soils with less than 15 ng/g. Ratios of p,p'-(DDD DDE)/DDT in soils under three land usages were: paddy field ＞ tree land ＞ fallow land, indicating that land usage inlfuenced the degradation of DDT in soils. Ratios of p,p'-(DDD DDE)/DDT ＞1, showing aged residues of DDTs in soils of the Taihu Lake region. The results were discussed with data from a former study that showed very low actual concentrations of HCH and DDT in soils in the Taihu Lake region, but according to the chemical half-lives and their concentrations in soils in 1980s, the concentration of DDT in soils seemed to be underestimated. In any case our data show that the ban on the use of HCH and DDT resulted in a tremendous reduction of these pesticide residues in soils, but there are still high amounts of pesticide residues in soils, which need more remediation processes.     

Sorption of 1-naphthol by plant cuticular fractions

The contribution of aliphatic-rich plant biopolymer to sorption of hydrophobic organic compounds is significantly important because of their preservation and accumulation in the soil environment,but sorption mechanism is still not fully understood.In this study, sorption of 1-naphthol by plant cuticular fractions was examined to better understand the contributions of respective fraction.Toward this end,cuticular materials were isolated from the fruits of tomato by chemical method.The tomato cuticle sheet consisted of waxes (6.5 wt%),cuticular monomer (69.5 wt%),and polysaccharide (24.0 wt%).Isotherms of l-naphthol to the cuticular fractions were nonlinear (N value (0.82-0.90)) at the whole tested concentration ranges.The KodKow ratios for bulk cuticle (TC1),dewaxed cuticle (TC2),cutin (TC4),and desugared cuticle (TC5) were larger than unity,suggested that tomato bulk cuticle and cutin are much powerful solption medium.Sorption capability of cutin (TC4) was 2.4 times higher than the nonsaponifiable fraction (TC3).The 1-naphthol interactions with tomato cuticular materials were governed by both hydrophobic-type interactions and polar (H-bonding) interactions. Removal of the wax and polysaccharide materials from the bulk tomato cuticle caused a significant increase in the sorption ability of the cuticular material.There was a linear negative trend between K_(oc) values and the amount of polysaccharides or fraction's polarities ((N O)/C);while a linear positive relationship between K_(oc) values and the content of cutin monomer (linear R~2=0.993) was observed for present in the cuticular fractions.Predominant sorbent of the hydrophobic organic compounds (HOCs) in the plant cuticular fraction was the cutin monomer,contributing to 91.7% of the total sorption of tomato bulk cuticle.