聚丙烯酰胺生物降解研究

研究了黄孢原毛平革菌对聚丙烯酰胺(PAM)的生物降解,从葡萄糖的加入量、pH、N浓度、Mn²⁺浓度和降解时间5个方面考察了对PAM降解的影响.结果表明,黄孢原毛平革菌对聚丙烯酰胺具有特殊的酶催化降解的能力,降解率可达50%,限氮条件(NH₄⁺=0.2g/L)和Mn2+浓度(Mn²⁺=0.017 5g/L)是菌株产聚丙烯酰胺降解酶的最佳条件.     

接种白腐菌堆肥修复五氯酚污染的土壤

采用4组堆肥装置A、B、C、D,分别往五氯酚污染土壤中不加外来菌、加入固定化黄孢原毛平革菌(Phanerochaete chrysosporium)、加入游离黄孢原毛平革菌及暴露在空气中不做任何处理.对其进行对比研究,探讨了利用堆肥的方法对五氯酚污染土壤进行生物修复的可行性.结果说明,添加黄孢原毛平革菌进行堆肥修复其效果要好于不加菌的效果,经过60d的堆肥,PCP基本得到降解,降解率都达到了94%以上,其中固定化菌又优于游离菌,固定化菌在第9d对PCP的降解率就达到了50%.此外,通过分析种子发芽率,有机质,微生物量碳等多项指标可以看出各组堆肥的腐熟情况都比较理想,为黄孢原毛平革菌降解五氯酚的过程提供了良好的外部环境,使土壤修复与堆肥能较好地结合起来.     

黄孢原毛平革菌反复脱色模拟染料废水的试验研究

研究了白腐真菌黄孢原毛平革菌在35℃、150r／min条件下培养3d后，培养基类型、染料浓度以及反复脱色次数对其脱色的影响。玉米芯浸出液是首选的可替代营养源的物质，培养3d的黄孢原毛平革菌6h脱色率即达92％。黄孢原毛平革菌能够在短时间(6h)基本脱色较高浓度(40mg／L，60mg／L)的活性艳红K-2BP模拟染料废水，对于低浓度(10mg／L，20mg／L)脱色作用则比较缓慢，浓度超过150mg／L即显示出对菌的毒性作用。玉米芯浸出液培养3d的菌可以重复脱色模拟废水6次以上，每批脱色率均在82％以上，但菌球的沉降性能变差。     

微量元素对黄孢原毛平革菌降解苯酚的影响

研究了氮源,铁、铜、锰等金属元素以及Tween 80等有机添加剂对黄孢原毛平革菌降解苯酚的影响。结果表明,氮源是黄孢原毛平革菌生长的必需元素,铜元素和锰元素对苯酚降解具有明显的促进作用,而铁元素的促进作用并不显著。各种微量元素的最佳浓度分别为:氮0.1 g/L、铁0.5 mmol/L、锰1.0 mmol/L、铜0.3 mmol/L、Tween 80 0.01 mmol/L。     

与黄孢原毛平革菌协同降解稻草的混合菌筛选

在纤维素筛选培养基中加入黄孢原毛平革菌稻草固态发酵的浸提液,用这种培养基从自然环境中筛选出与黄孢原毛平革菌相容的菌株,在PDA和MEA平板上进行相容性测试,选择出生长稳定,可以与黄孢原毛平革菌较好共存的6株真菌.考察了6株真菌分别与黄孢原毛平革菌混合培养降解稻草时的酶活和对木质纤维素类组分的降解,筛选出3组更优的组合,其混合培养时,对于纤维素酶能产生较好的协同效果,纤维素降解效果较好.对于木质素降解效率的显著提高,可能是由于发酵体系内的胞外酶和一些活性因子的作用.     

黄孢原毛平革菌对6种染料的脱色降解

在黄孢原毛平革菌与6种染料的液体静培养体系中,不同浓度的染料均发生脱色降解。共培育30 d时,刚果红、直接冻黄G和活性翠蓝KN－G达到92%～99％的脱色率。10,50,100 mg/L活性翠蓝KN－G,50,100 mg/L金莲橙O与天青蓝A,100 mg/L活性艳蓝KN－R的降解率达70%以上;18个样品中60%的降解率超过50%。生物吸附和生物降解是2个重要过程。研究表明,黄孢原毛平革菌对各种染料类群具有广谱有效的脱色降解能力。      

真菌对磺胺二甲氧嘧啶降解过程的转录组分析及差异表达基因的功能分析

采用黄孢原毛平革菌降解磺胺二甲氧嘧啶（SDM）,4 d后SDM的去除率达74%,降解速率达3.24 mg·L^-1·d^-1.采用Illumina高通量测序平台对降解SDM后的菌株与对照组菌株进行测序,通过GO和KEGG富集分析,得到的差异表达基因能显著富集到与降解相关的“氧化还原酶活性”途径,及与应激反应有关的“ABC转运体”路径.通过对高表达高上调的差异表达基因的筛选与分析,得到了耐受和降解SDM的功能基因.水通道蛋白、ABC转运蛋白及甲基转运酶基因等在黄孢原毛平革菌对环境的耐受中起到重要的作用.乙醇氧化酶、细胞色素P450和糖苷水解酶等在黄孢原毛平革菌对SDM的降解中起着关键的作用.本文从转录组水平分析了SDM的降解机制,为黄孢原毛平革菌在环境修复中的应用提供一定的参考.     

AFLP法分析染料对黄孢原毛平革菌的损伤效应

利用AFLP对DNA碱基序列分辨率高的特性,研究纯培养条件下染料对黄孢原毛平革菌(P. chrysosporium)菌株DNA的损伤效应.在对黄孢原毛平革菌(P. chrysosporium)AFLP最佳引物筛选的基础上,通过分析最佳引物扩增结果中DNA相似性、染料浓度与DNA相似性关系以及构建UPGMA相似性树状图,评价染料对P. chrysosporium DNA损伤效应.结果表明, AFLP法可较好反映染料对DNA损伤效应,在优选的3种引物组合中, E-AAC/M-CAA引物组合AFLP扩增效果最好,共获得35条DNA扩增条带,其中有12条具有特异性,多态性比率34.3%;在进行DNA相似性分析和UPGMA法聚类分析后发现, AFLP技术较好地表达了DNA相似性与染料浓度的正相关关系,并且显示P. chrysosporium对染料损伤效应敏感,染料浓度不宜高于50mg/L,当染料浓度达到300mg/L即可对P. chrysosporium产生较大损伤作用,使之基本丧失原有的处理功能.     

染料浓度和盐度对白腐真菌同工酶的电泳分析

应用聚丙烯酰胺凝胶电泳技术分析,并比较了染料废水的两大因子(染料浓度和盐度)对黄孢原毛平革菌过氧化物同工酶谱带的影响程度,结果表明:在染料或盐胁迫下,黄孢原毛平革菌均表现出应急产酶的响应,虽然过氧化物同工酶谱带数量未发生增加或减少,但谱带强度受到较大影响,活性艳红X-3B浓度≤200mg/L或NaCl浓度≤15g/L时均能诱导过量产酶,继续提高染料浓度或盐度将导致产酶抑制,染料浓度对酶带的影响作用高于盐度。     

黄孢原毛平革菌反应体系中藜芦醇的动态变化

用紫外分光光度法测定黄孢原毛平革菌(Phanerochaete chrysosporium)反应体系中藜芦醇(3,4-二甲氧基苯甲醇,VA)的动态变化,以吸收曲线波型的变化,分析VA的降解;根据培养液在特征最大吸收波长处吸光度的变化,计算VA量的动态变化.研究表明,在菌的反应体系中,细胞自身内源性VA的合成反应,导致总VA量短时间的增加;但外源VA发生逐步的降解,15d内已基本消失;菌的可作用底物染料的存在有助于内源性VA的合成.低浓度的苯甲醇(BA)对黄孢原毛平革菌的降解反应具有促进作用,BA完全可以代替VA的作用.     

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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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.     

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.     

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.     

Alterations of organ histopathology and metallolhionein mRNA expression in silver barb, Puntius gonionotus during subchronic cadmium exposure

Common silver barb,Puntius gonionotus,exposed to the nominal concentration of 0.06 mg/L Cd for 60 d,were assessed for histopathological alterations(gills,liver and kidney),metal accumulation,and metallothionein(MT)mRNA expression.Fish exhibited pathological symptoms such as hypertrophy and hyperplasia of primary and secondary gill lamellae,vacuolization in hepatocytes,and prominent tubular and glomerular damage in the kidney.In addition,kidney accumulated the highest content of cadmium,more than gills and liver.Expression of MT mRNA was increased in both liver and kidney of treated fish.Hepatic MT levels remained high after fish were removed to Cd-free water.In contrast,MT expression in kidney was peaked after 28 d of treatment and drastically dropped when fish were removed to Cd-free water.The high concentrations of Cd in hepatic tissues indicated an accumulation site or permanent damage on this tissue.