高效降酚菌Bacillus sp.JY01的固定化及降解特性研究

以海藻酸钠作为载体将降酚菌株Bacillussp.JY01进行固定化包埋,并通过正交实验确定了该菌株固定化细胞制备的最优条件;研究并对比了固定化细胞和游离细胞的降酚性能,研究结果表明最佳固定条件为:海藻酸钠质量分数3%菌液量:海藻酸钠水溶液体积比4:30、氯化钙含量为3%、钙化交联时间8h;固定化细胞降解苯酚的最适温度是32℃,最适pH值范围为7.0～7.5,最适条件下能高效降解质量分数为1300mg/L的苯酚溶液,固定化细胞重复利用8次苯酚降解率仍可达到96.8%,该固定化细胞降酚性能优于游离细胞。这将为该细菌进一步应用于含酚工业废水的生物处理提供可靠的控制条件。     

PVA固定化Corynebacterium sp.JY03降解苯酚的特性研究

以PVA(聚乙烯醇)作为载体将降酚菌株Corynebacterium sp.JY03进行固定化包埋处理,正交试验确定该菌株固定化细胞制备的条件,然后对固定化细胞的降酚性能进行研究。试验确定最佳固定条件为:PVA质量分数为6%,菌液量/PVA水溶液体积比为6/30,氯化钙含量为2.0%,钙化交联时间为8 h;固定化细胞降解苯酚的最适温度为30~35℃,最适pH值为6.5~7.5,在初始苯酚浓度为700 mg/L,装液量50 mL,培养42 h后,苯酚降解率达99.1%。固定化细胞重复利用6次苯酚降解率仍高于85.2%,其性能优于游离细胞,这将为该菌株进一步应用于含酚废水的生物处理提供实践条件。     

固定化条件对苯系物细胞传感器检测效果的影响

利用海藻酸钠作为包埋剂对苯系物细胞传感器Pseudomonas fluorescens A506(pTS)进行固定化处理,并对细胞浓度、固定化时间、投加量等检测因素进行了优化.固定化后的细胞与游离细胞对苯系污染物的检测效果进行对比.经过2 h固定化处理的细胞传感器,在检测时间1.5～6.0 h内,信号上升速度为游离细胞的2.26倍,信号最大值为游离细胞的2.23倍.固定化处理后的细胞生长缓慢,且浓度远低于游离细胞,说明了固定化处理后的单位细胞信号强度增加.同时,细胞浓度随着固定化时间的延长而降低.在影响因素方面,细胞浓度和固定化时间对于检测信号的影响较大.固定化后,对高浓度的苯系污染物有着更快的信号响应.     

一株产碱杆菌DN25中降氰酶的提取和特性研究

从实验室自行保藏的一株高效降氰菌株——产碱杆菌Alcaligenes sp.DN25中提取降氰酶,并研究其降解特性。通过分别测定胞内酶、胞外酶和细胞碎片对氰化物的降解率,初步判断该菌株降氰酶主要分布在细胞内。所提取的降氰酶具有较好的保存稳定性和pH稳定性,降解最适条件为温度35℃、pH7.0,并测得该降氰酶的米氏常数Km和最大反应速率分别为267.8mg/L和6.71mg/(L·min)。检测出该酶降解氰化物的产物之一为NH3,降解完全;尿素对酶活有抑制作用,初步可断定此酶中存在氰水合酶。     

丝瓜络固定化黄曲霉A5p1及其脱色糖蜜酒精废水

采用丝瓜络作为固定化材料,研究固定化黄曲霉(Aspergillus flavus)A5p1对糖蜜酒精废水的脱色条件。结果表明,固定化细胞的最优脱色条件为pH 5.0,温度40℃,转速150 r/min,接种量4.3×104spores/mL。固定化可促进细胞生长,菌株的最大产H2O2速率和脱色率增加,对2%浓度(色度值A475≈3.5)糖蜜酒精废液的4 d脱色率由游离细胞的53.0%提高至65.7%。构建固定化细胞反应器连续处理糖蜜酒精模拟废水具有一定的可行性,反应器可稳定运行约25 d,前18 d可保持50%的脱色率。     

己烯雌酚降解菌固定化条件优化及其降解性能

以海藻酸钠为包埋剂,制备了己烯雌酚(DES)降解菌株S(Serratia sp.)的固定化菌剂,通过正交试验确定了菌株S的最佳固定化条件:海藻酸钠质量分数为4%,菌悬液与海藻酸钠重量比为1:2,CaCl₂质量分数为4%,交联时间为6h.菌株S的固定化菌剂比游离菌株更有优势;7d时,固定化菌株S对无机盐培养基中DES的去除率为83.84%,较游离菌株高22.84%.优化了固定化菌株S对溶液中DES的处理条件:固定化菌株S接入量为300g/L,初始底物浓度为20~40mg/L.固定化菌株S对DES浓度为40.01,37.90 ,33.52μg/L 3个排污口污水中DES的去除率分别达90.0%,96.0%,96.7%.利用固定化菌株处理实际污水中DES等雌激素具有应用潜力.     

固定化优势菌种降解2,6-二叔丁基酚

 用海藻酸钙包埋固定优势降解菌(Alcaligenes sp.)降解 2,6-二叔丁基酚(2,6-2DTBP).结果表明,菌株经固定化包埋后,降解底物 2,6-DTBP的能力大大提高,在 100.0mg/L 的初始浓度下其降解率在 12d 可达到 86%.与未固定菌株相比,固定化菌株对 pH 值和温度的适应范围更宽,对底物具有更高的降解能力.对固定化菌株的降解反应过程进行动力学分析,该降解反应符合一级动力学特征,其动力学常数为 0.1519,半衰期为 4.56d.扫描电镜观察到菌种在海藻酸钙包埋载体中能良好地生长和繁殖.     

改良型固定化Pseudomonas oleovorans DT4降解四氢呋喃的研究

成功构建出了新型的海藻酸钠-活性炭纤维复合固定化Pseudomonas oleovorans DT4来去除四氢呋喃,并优化了该复合固定化载体组分的含量,发现在海藻酸钠含量为3%,活性炭纤维含量为1.5%条件下,制备成细胞浓度为6×109g-1的协同固定化细胞在初始THF浓度为360 mg·L-1时的降解速率达到最大为24.0 mg·(L.h)-1,同时其机械强度也得到了显著的提升.此外与游离菌体相比,该固定化细胞在不同温度和pH条件下的降解速率表现得更加稳定,且去除效果均能稳定在80%以上.在改进的反应体系中,协同固定化小球高效降解THF的重复利用次数达到80批以上,显示出该载体良好的可行性.     

镰刀菌12号固定化细胞降解氰的研究

在镰刀菌12号的培养基中添加氰化物作诱导剂,可显著地提高酶活力,经海藻酸钙固定后相对活性为89.66%。 比较了自然细胞与固定化细胞的某些性质,两种细胞反应的最适温度分别为25—30℃和35—45℃。反应最适pH值为8.0—9.0。固定化细胞较自然细胞热稳定性明显增加。 固定化细胞柱连续处理浓度为500ppm和1000ppmCN-,流速分别为30ml/h和15ml/h,当进水CN-500ppm,连续运转90h,出水CN-<10ppm。     

聚乙烯醇-海藻酸钠-活性炭固定化菌球处理二氯甲烷的研究

本研究以聚乙烯醇-海藻酸钠-活性炭为复合载体、自行筛选的高效降解菌Methylobacterium rhodesianum H13为目标菌制备固定化小球,用于去除二氯甲烷,同时优化了固定化小球的制备条件.结果表明,当聚乙烯醇浓度为8%,海藻酸钠浓度为5%,CaCl₂浓度为2%,活性炭浓度为1%,包菌量为0.6 g,钙化时间为8 h时,复合固定化细胞对DCM的降解速率达到最高为18.2 mg·L^-1·h^-1,同时活性炭的添加对小球的机械强度也有显著提升.与游离细胞相比,固定化细胞具有更好的热稳定性与pH稳定性,并且固定化细胞的重复利用性较好,连续处理9批底物后,DCM降解速率仍保持在13.4 mg·L^-1·h^-1以上.吸附动力学与等温线拟合结果显示,在固定化载体中加入1%活性炭后,小球对DCM的吸附量明显提高,而且吸附能在更短的时间内达到平衡.吸附动力学符合准二级反应动力学方程,复合载体对DCM的吸附遵循Langmuir吸附模型（R²>0.99）.     

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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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.