镰刀菌(Fusarium sp.)HJ01对中性艳蓝GL的脱色降解特性研究

利用镰刀菌(Fusarium sp.)HJ01降解中性艳蓝GL,经扩增翻译延伸因子TEF1对HJ01菌株做了种的鉴定,并考察了温度、pH值对该菌株降解酶漆酶的活性,以及对中性艳蓝GL脱色率及COD去除率的影响.结果表明,该菌株属层出镰刀菌(Fusarmm proliferatum).HJ01菌株所产降解酶漆酶在pH为2.0～8.0内相对活性在12%～96%之间,在pH=4.0时,相对活性最大;在25～65 ℃内,随着温度的升高,漆酶相对活性增大;在65～85 ℃内,随着温度增大,漆酶相对活性降低.HJ01菌株在25℃、50 r·min-1条件下,于液体培养基中培养96h后加入到25mL含100mg·L-1中性艳蓝GL的溶液中,在35℃、50 r·min-1、pH=4.0条件下继续培养100h后,中性艳蓝GL的脱色率达100%,培养至120h后,COD去除率达87.8%.紫外光谱分析表明,中性艳蓝GL的发色基团蒽醌环被漆酶打破.     

Trametes sp.LS-10C产漆酶发酵培养基优化及其漆酶对偶氮染料的脱色性能

为了提高菌株Trametes sp.LS-10C的漆酶产量并初步研究该酶对偶氮染料的脱色性能,本文通过单因素实验及均匀设计对菌株Trametes sp.LS-10C产漆酶培养基进行了优化,获得最优培养基配方为:葡萄糖14.4 g·L~(-1)、豆渣10.1 g·L~(-1)、麸皮100 g·L~(-1)、NH_4Cl_3g·L~(-1)、KH2PO41.4g·L~(-1)、CuSO_4’5H2O 1 g·L~(-1)、NaCl 1 g·L~(-1)、MgSO_40.8 g·L~(-1)、CaCl_20.5 g·L~(-1).菌株发酵至8 d时漆酶产量为595.15 U·m L~(-1),约是优化前的35.66倍.进一步研究表明,该漆酶-介体系统对酸性红GR、酸性蓝40和酸性铬蓝K等3种偶氮染料脱色10 h后脱色率分别高达96.86%、91.28%和86.31%.相比公开发表的文献,本研究所报道的Trametes sp.LS-10C利用优化培养基发酵产漆酶达到了较高水平,具有酶活力高和发酵时间短等特征,且该漆酶在处理偶氮染料废水脱色领域中具有进一步研发和应用价值.     

多环芳烃降解菌的分离鉴定及其生理特性研究

从兰州石化污水处理厂的污泥中分离出一株能分别以萘、蒽为唯一碳源生长的细菌(BDP01). 通过菌株形态观察及16S rDNA序列比对, 确定该菌株属于假单胞菌属的绿脓杆菌(Pseudomonas aeruginosa). 菌株在LB培养基中48 h内呈对数生长, 48 h到72 h进入生长稳定期, 72 h后开始进入衰亡期;其最佳培养温度为30 ℃, 最适生长pH为中性或弱碱性, 该菌能在萘和蒽的质量浓度分别为100 mg·L^-1和50 mg·L^-1的无机盐培养基中良好生长, 与已报道的其他微生物比较, BDP01在较短时间内具有较强的降解多环芳烃能力, 1%的接种量15 d内可降解89.64%的萘和77.21%的蒽. 采用PCR和琼脂糖凝胶电泳技术检测到菌株基因组中有邻苯二酚2,3-双加氧酶 (C23O)基因. 测序结果与NCBI数据库发布的C23O基因序列相似度为92%. 该酶在20~65 ℃以邻苯二酚为底物时, 细胞裂解液中酶活力变化范围为13.10~540.86 U.     

Citrobacter sp. LW-3对偶氮染料甲基橙的降解脱色特性研究

从长期受染料污染的土壤中分离出一株能对甲基橙高效降解脱色的柠檬酸盐杆菌Citrobacter sp.LW-3.菌株LW-3在添加了0.5%(g/100 mL)葡萄糖的MSM-1培养基中,16 h使100 mg·L-1甲基橙降解掉90.56%.该菌株对甲基橙降解脱色可在完全好氧条件下,脱色与木质素过氧化物酶、NADH-DCIP原酶、核黄素还原酶有关.菌株LW-3降解甲基橙的适宜温度较广,20~40℃24 h均能使超过80%的甲基橙降解脱色,适宜pH范围为6.0~8.0.通气量对菌株LW-3影响较小,24 h各装液量组(25~200 mL)甲基橙均能降解近90%.同时,菌株LW-3具有较广的染料降解谱,3 d内可使20 mg·L-1另3种偶氮染料、两种三苯甲烷类染料和一种蒽醌类染料很好脱色.菌株LW-3在染料废水的生物处理方面有较强的实际应用价值.     

久效磷降解菌的分离及其酶促降解特性研究

从某农药厂废水处理池的污泥中分离到1株久效磷高效降解菌株M-1,经过对其形态特征、生理生化、以及16S rDNA序列分析,该菌株初步鉴定为Paracoccus sp..M-1能以久效磷作为唯一碳源生长,24 h对100 mg·L^-1久效磷的降解效率为92.47%.久效磷降解酶定域表达试验表明该酶为胞内酶,组成表达.久效磷酶促降解的最适反应pH为8.0,最适反应温度为25 ℃;其米氏常数（K_m）为0.29 μmol·mL^-1,最大降解速率(V_max)为682.12 μmol·(min·mg)^-1.久效磷降解酶热稳定性差,碱性条件下能够保持较高降解活性.     

菌株Sphingomonas sp. FL降解溴氨酸的特性研究

分离了1株澳氨酸降解菌.其可以溴氨酸为唯一碳源进行降解并使其脱色,通过16S rRNA基因序列比较和生理生化特性分析,将其归为鞘氨醇单胞菌属.溴氨酸降解和菌株生长的最适条件为:温度30℃,pH 7.0,摇床转速100 r/rain,(NH4)2SO4作为氮源,在此条件下,溴氨酸(100 mg/L)在14 h内的脱色率可达99%.低浓度NaCl(<2%)对脱色有促进作用,而高浓度NaCl(≥2%)对脱色产生抑制.以Haldane底物抑制模型表征溴氨酸初始浓度对脱色的影响.确定当初始浓度为1 393.5 mg/L时可取得最佳比降解速率1.4 h-1.菌株不能将溴氨酸完全矿化,至反应终点52.4%的有机碳得到去除.利用GC-MS和HPLC-MS分析代谢产物显示.溴氨酸降解的中间产物是邻苯二甲酸,终产物可能为2-氨基-3-羟基-5-溴苯磺酸或2-氨基-4-羟基-5-溴苯磺酸,邻苯二甲酸可经3,4-二羟基苯甲酸途径进一步降解而被菌体利用.     

利用固定化技术与添加表面活性剂增强双酚A降解率的研究

以SiO_2为载体,戊二醛为交联剂,用以固定游离态漆酶降解双酚A(Bisphenol A,BPA).同时,对比了游离态漆酶与固定化漆酶在不同pH值与温度下的最佳酶活与酶稳定性,再通过添加表面活性剂来增加酶活及BPA去除效果.结果表明,与游离态漆酶相比,固定化漆酶对不同酸碱的适应性与耐热性均有所提高,在pH为2~8时,固定化漆酶的活性均高于游离态漆酶,且最佳活性温度也从游离态漆酶的40℃提高至50℃.在BPA降解率方面,固定化漆酶的降解率皆高于游离态漆酶,反应6 h后,对100 mg·L~(-1)BPA的去除率可分别达到92%与85%.通过添加非离子型表面活性剂Triton X-100,反应4 h后固定化漆酶可完全降解BPA,相对活性为67%;游离态漆酶于6 h将BPA降解完全,相对活性为32%.实验证实Triton X-100可作为酶活诱导剂用以提高漆酶活性及BPA的降解率,降低了处理成本且具有一定的应用前景.     

菲降解菌株GY2B的分离鉴定及其降解特性

对一株菲降解菌进行了鉴定并对其降解特性进行了研究.初步鉴定菌株GY2B为鞘氨醇单胞菌(Sphingomonassp.).菌株GY2B有较高的降解效率,当无机盐培养液中菲初始浓度为100mg/L时,48h内对菲降解率达到99.1%.添加100mg/L的葡萄糖和蛋白胨均可以促进菌株的生长和菲的降解,pH值为中性条件时对细胞的生长较为有利.GY2B菌株还能降解1-羟基-2-萘酸、2-萘酚、萘、水杨酸、邻苯二酚和苯酚等多种芳香化合物并利用其为碳源和能源生长繁殖.GY2B菌株对菲的降解可能通过水杨酸途径.     

黄曲霉A5p1对偶氮染料的降解特性和降解途径

以黄曲霉菌株A5p1为生物材料,研究其脱色染料的广谱性,并选择偶氮染料直接蓝71（DB71）为模型底物,探讨脱色特性及降解产物.该菌株对15种染料的脱色测试结果表明,染料浓度为100mg/L时脱色效率为61.7%~100%.该菌对偶氮染料DB71具有生物吸附和生物降解的双重作用,在pH值7.0,温度30℃,染料浓度300mg/L,蔗糖为碳源时对DB71 脱色率为100%.酶分析显示葡萄糖氧化酶和锰过氧化物酶参与染料的降解.FTIR、GC-MS和LC-MS分析确定代谢终产物为萘胺、叠氮萘、2-羟基-6-草酰-苯甲酸和1-萘酚.     

菌株Sphingomonas sp. FL降解溴氨酸的特性研究

分离了1株溴氨酸降解菌,其可以溴氨酸为唯一碳源进行降解并使其脱色,通过16S rRNA基因序列比较和生理生化特性分析,将其归为鞘氨醇单胞菌属.溴氨酸降解和菌株生长的最适条件为：温度30℃,pH 7.0,摇床转速100 r/min ,(NH₄)₂SO₄作为氮源,在此条件下,溴氨酸（100 mg/L）在14 h内的脱色率可达99%.低浓度NaCl（<2%）对脱色有促进作用,而高浓度NaCl（≥2%）对脱色产生抑制.以Haldane底物抑制模型表征溴氨酸初始浓度对脱色的影响,确定当初始浓度为1 393.5 mg/L 时可取得最佳比降解速率1.4 h^-1.菌株不能将溴氨酸完全矿化,至反应终点52.4%的有机碳得到去除.利用GC-MS和HPLC-MS分析代谢产物显示,溴氨酸降解的中间产物是邻苯二甲酸,终产物可能为2-氨基-3-羟基-5-溴苯磺酸或2-氨基-4-羟基-5-溴苯磺酸,邻苯二甲酸可经3,4-二羟基苯甲酸途径进一步降解而被菌体利用.     

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.     

Effects of two sludge application on fractionation and phytotoxicity of zinc and copper in the soil

The potential harm of heavy metals is a primary concern in application of sludge to the agricultural land. A pot experiment was conducted to evaluate the effect of two sludges on fractionation of Zn and Cu in soil and their phytotoxicity to pakchoi. The loamy soil was mixed with 0%, 20%, 40%, 60% and 80% (by weight) of digested sewage sludge (SS) and composted sludge (SC). The additions of both sludges caused a significant raise in all fractions, resulting in that exchangeable (EXCH) and organic bound (OM) became predominance of Zn and organic bound Cu occupied the largest portion. There was more available amount of Zn and Cu in SS treatments than SC treatments. During the pot experiment, the concentration of Zn in EXCH, carbonate (CAR) and OM and Cu in EXCH and OM fractions decreased in all treatments, so their bioavailability reduced. Germination rate and plant biomass decreased when the addition rate was high and the best yield appeared in 20% mixtures at the harvest of pakchoi. The two sludges increased tissue contents of Zn and Cu especially in the SS treatments. Zn in pakchoi was not only in relationship to ΔEXCH and ΔCAR forms but also in ΔOM forms in the sludge-soil mixtures. Tissue content of Cu in pakchoi grown on SC-soils could not be predicted by ΔEXCH. These correlation rates between Zn and Cu accumulation in pakchoi and variation of different fractions increased with time, which might indicate that sludges represented stronger impacts on the plant in long-term land application.     

Intraspecific differences in effects of co-contamination of cadmium and

A hydroponic experiment was carried out to study intraspecific differences in the effects of different concentrations of cadmium (Cd)(0-10 mg/L) and arsenate (As(V)) (0-8 mg/L) on the growth parameters and accumulation of Cd and As in six wheat varieties Jing-9428, Duokang-1, Jingdong-11, Jing-411, Jingdong-8 and Zhongmai-8. The endpoints of wheat seedlings, including seed germination,biomass, root length and shoot height, decreased with increasing the Cd and As concentrations. Significant differences in seed germination, biomass, root length, shoot height and the accumulation of Cd and As were observed between the treatments and among the varieties (p ＜ 0.05). The lethal dosage 50% were about 20, 80, 60, 60, 80 and 20 mg As/L for Jing-9428, Duokang-1, Jingdong-11,Jing-411, Jingdong-8 and Zhongmai-8, respectively, and the corresponding values for Cd were about 30, 80, 20, 40, 60 and 10 mg Cd/L, respectively. Among the six varieties, Duokang-1 was found to be the most resistant to Cd and As toxicity, and Zhongmai-8 was the most sensitive to Cd and As co-contamination. The resistance of the six varieties was found dependant on the seedling uptake of Cd and As. Duokang-1 was the most suitable for cultivation in Cd and As co-contaminated soils.     

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.