零价铁对2,4-二氯酚生物还原脱氯的影响研究

采用间歇试验,接种驯化两月的厌氧混合微生物,考察厌氧体系中添加零价铁（Fe^0）对2,4-二氯酚（2,4-DCP）生物还原脱氯效果的影响,并对影响“Fe^O＋微生物”体系的一些因素进行了探索。结果显示：与零价铁或微生物的单独作用相比,“Fe^O＋微生物”体系能够有效促进2,4-DCP的脱氯反应,最佳Fe^O投加量和微生物接种量分别为0.5g／L和376.2mgVSS／L;初始pH=8.0对2,4-DCP的转化效果最好,偏酸性环境不利于污染物转化;微生物接种量与铁用量之间有一适宜比例,一定范围内增加微生物接种量可催生出更多可降解污染物的酶或酶系,提高2,4-DCP的降解效果。     

不同电子供体对2,4-二氯酚还原脱氯的影响

以葡萄糖、乙酸钠、Fe0、Fe0 葡萄糖、Fe0 乙酸钠作为电子供体,接种未驯化厌氧混合菌,考察2,4-二氯酚(2,4-DCP)的还原脱氯特性及Fe0作为电子供体的最佳作用条件与持续性特征.结果表明:与葡萄糖的作用相比,Fe0 葡萄糖可有效提高目标物脱氯效果;乙酸钠、Fe0及Fe0 乙酸钠均为有效电子供体,其中Fe0作为电子供体时目标物脱氯效果最佳,最佳作用条件为初始pH8.0,Fe0投加量2.0 g/L,4-CP为其主要脱氯中间产物;Fe0可持续供给2,4-DCP还原脱氯所需电子,而乙酸钠不断消耗后其脱氯效果与Fe0作为电子供体有明显差距.     

降解2,4-二氯酚微生物的分离及其2,4-二氯酚羟化酶基因的克隆和表达

从土壤中分离到一株降解2,4-二氯酚能力较强的细菌菌株GT241-1,经鉴定该菌株属于假单胞菌属。菌株GT241-1在最适条件下能在48h内将90mg/L的2,4-DCP降解91%,能利用2,4-二氯酚、2,4-二氯苯氧乙酸、苯甲酸和儿茶酚为唯一碳源生长。采用Southern杂交对2,4-二氯酚羟化酶基因（dcpA）定位后构建基因组文库,再用斑点杂交筛选目的转化子,克隆了该菌株的dcpA。序列测定得知含dcpA的亚克隆片段全长2389bp,其中dcpA基因编码区1797bp。核苷酸和氨基酸序列分析表明,dcpA与已在GenBank登记的相关基因有一定的差异。dcpA基因能够在大肠杆菌转化子中成功地表达有生物活性的酶。     

固定化白腐真菌降解水溶液中2,4-二氯酚的研究

研究固定化黄孢原毛平革菌对水溶液中2,4-二氯酚（2,4-DCP）的降解效果,探讨固定化黄孢原毛平革菌处理水溶液中氯酚类污染物的可行性．结果表明,采用固定化方法处理的白腐真菌．其产酶稳定性及酶活均比游离态白腐真菌有显著提高．2,4-DCP降解效果受固定化孢子接种量、pH值、摇床转速、2,4-DCP的初始浓度和表面活性剂浓度的影响．当pH为4．5,摇床转速180r／min,培养基含有1％的Tween 80,2,4-DCP初始浓度为40mg／L时,加入10mL固定化白腐真菌孢子,2,4-DCP去除效果最好．     

纳米零价铁耦合假单胞菌协同高效降解五氯苯

【背景】氯苯类化合物广泛应用于工业化合物的生产,由于其具有高毒性和环境持久性的特点,对人类健康和生态环境造成严重威胁,寻找高效降解这类化合物的新方法成为研究热点。【目的】将纳米零价铁与假单胞菌耦合,探究其在好氧条件下对五氯苯的降解效果和降解机理。【方法】建立纳米零价铁和假单胞菌降解五氯苯的反应体系,通过测定各反应体系中五氯苯及其中间产物的浓度,以及观测细菌的生长状况变化等,分析反应体系的降解效果及其可能的降解机理。【结果】纳米零价铁耦合假单胞菌的反应体系相对于两者的单一体系表现出更好的降解效果,36h的降解率可达55.4%,反应过程符合伪一级反应动力学,速率常数为0.02048h~(-1)。根据GC-MS所测的中间产物推测,该体系的反应机理为纳米零价铁在好氧条件下反应产生羟基自由基,攻击五氯苯使其变为低氯苯类化合物,假单胞菌进一步利用这些低氯苯类化合物;同时,假单胞菌又为纳米零价铁提供附着位点,有效地降低了纳米零价铁的聚集性,提高了反应活性。【结论】研究建立的纳米零价铁耦合假单胞菌反应体系对五氯苯具有较好的降解效果,为含有高氯代苯类等有机污染物的复杂环境的修复提供参考。     

不同价态铁元素对厌氧微生物降解2,4,6-三氯酚的影响

【目的】考察初始pH值为5.0?10.0时, 不同价态铁元素(Fe0、Fe2+和Fe3+)对厌氧微生物降解2,4,6-三氯酚(2,4,6-TCP)的影响。【方法】采用间歇试验, 接种驯化3个月的厌氧污泥, 向其中分别投加Fe0、Fe2+和Fe3+, 测定体系中2,4,6-TCP浓度、pH值、铁离子浓度和微生物脱氢酶活性。【结果】“Fe0/Fe2+/Fe3+-微生物”体系对2,4,6-TCP的降解效率, 在初始pH值为中性偏酸性时, “Fe2+-微生物”体系>“Fe0-微生物”体系>“Fe3+-微生物”体系; 而当初始pH值为碱性时, “Fe0-微生物”体系>“Fe2+-微生物”体系>“Fe3+-微生物”体系; “Fe0/Fe2+/Fe3+-微生物”三种体系均有调节pH值的能力, 其中“Fe0-微生物”体系调节能力最强; 在不同初始pH值条件下不同价态铁元素对厌氧微生物活性的影响结果与其对2,4,6-TCP的影响规律基本相同。【结论】不同价态铁元素对厌氧微生物降解2,4,6-TCP的影响与初始pH值、体系实时pH值和铁元素价态及浓度等因素有关。     

酿酒酵母B5不对称还原制备手性药物中间体R-2′-氯-1-苯乙醇的研究

从 11株微生物中筛选出 4株具有不对称还原 2′ 氯 苯乙酮能力的酵母 ,其中酿酒酵母B5的还原产率与对映体选择性最佳。确定了酿酒酵母B5对 2′ 氯 苯乙酮还原的最佳反应时间为 2 4h ;最佳pH 8 0 ;最佳反应温度为2 5℃ ;最佳共底物为 5 % (体积比 )乙醇。同时研究了底物浓度、微生物的量、微生物的培养条件等对反应产率和立体选择性的影响。细胞浓度为 10 75mg mL(细胞干重 反应体积 )的酿酒酵母B5可将 6 47mmol L的 2′ 氯 苯乙酮10 0 %地转化为R 2′ 氯 1 苯乙醇 ,其对映体选择性为 10 0 %。酿酒酵母B5可重复利用的特点可提高产物的产量。     

假单胞菌ZWL73降解4-氯硝基苯的代谢途径研究

氯代硝基芳香烃是一类环境中难以降解的有毒污染物。一株高效分解4-氯硝基苯的假单胞菌分离于4-氯硝基苯污染土壤, 可以完全降解4-氯硝基苯, 并以之为C源、N源生长。为阐明其降解4-氯硝基苯的代谢途径, 通过对以底物生长的降解菌的酶学分析, 检测到其还原降解的两个关键酶即初始酶硝基还原酶和苯环开环酶2-氨基-5-氯酚1, 6-双加氧酶的活性; 结合其它检测如培养液中降解产物分析、相关底物生长实验结果, 确定了其降解途径是通过部分还原途径。     

酿酒酵母B5不对称还原制备手性药物中间体R-2′-氯-1-苯乙醇的研究

从11株微生物中筛选出4株具有不对称还原2′-氯-苯乙酮能力的酵母,其中酿酒酵母B5的还原产率与对映体选择性最佳。确定了酿酒酵母B5对2′-氯-苯乙酮还原的最佳反应时间为24h;最佳pH 8.0;最佳反应温度为25℃;最佳共底物为5%（体积比）乙醇。同时研究了底物浓度、微生物的量、微生物的培养条件等对反应产率和立体选择性的影响。细胞浓度为10.75mg/mL（细胞干重/反应体积）的酿酒酵母B5可将647mmol/L的2′-氯-苯乙酮100%地转化为R-2′-氯-1-苯乙醇,其对映体选择性为100%。酿酒酵母B5可重复利用的特点可提高产物的产量。     

2,4-二氯苯酚在土壤与河流底泥中降解动力学

以南京化学工业园内四柳河沿岸土壤与河流底泥为研究对象,通过土壤灭菌、温度与污染物初始浓度调控,研究了2,4-二氯苯酚在土壤与河流底泥中降解动力学及其影响因子。结果表明:微生物对2,4-二氯苯酚降解起主导作用,在45d内,非灭菌土壤和河流底泥的降解率分别是灭菌条件下的1.5～3倍、1.4～2.8倍,土壤和河流底泥中的2,4-二氯苯酚微生物降解量分别为0.128～0.599和0.113～0.718mg·kg-1,非灭菌处理半衰期时间短于灭菌处理;(10±1)℃～(30±1)℃范围内,随着温度的增高,2,4-二氯苯酚降解加快,在(30±1)℃土壤与河流底泥中残留量最小,分别为0.305和0.203mg·kg-1,半衰期也最短;土壤与河流底泥中的2,4-二氯苯酚均在其浓度为0.5mg·kg-1时降解最快,随着初始浓度的增加,2,4-二氯苯酚降解速度呈现递减趋势,半衰期增长。     

Long-Term Sustainability of Reductive Dechlorination Reactions at Chlorinatedsolvents Sites

The sustainability of biodegradation reactions is of interest at Type 1 chlorinated solvent sites where monitored natural attenuation is being considered as a remedial alternative. Type 1 chlorinated solvent sites are sites undergoing reductive dechlorination where anthropogenic substrates (such as landfill leachate or fermentable organics in the waste materials) ferment to produce hydrogen, a key electron donor. A framework is provided that classifies Type 1 chlorinated solvent sites based on the relative amounts and the depletion rates of the electron donors and the electron acceptors (i.e., chlorinated solvents). Expressions are presented for estimating the total electron donor demand due to the presence of solvents and competing electron acceptors such as dissolved oxygen, nitrate, and sulfate. Finally, a database of 13 chlorinated solvent sites was analyzed to estimate the median and maximum mass discharge rate for dissolved oxygen, nitrate, and sulfate flowing into chlorinated solvent plumes. These values were then used to calculate the amount of hydrogen equivalents and potential for lost perchloroethylene (PCE) biodegradation represented by the inflow of these competing electron acceptors. The median and maximum mass of PCE biodegradation lost due to competing electron acceptors, assuming 100% efficiency, was 226 and 4621 kg year(-1), respectively.     

Reductive Dechlorination of Tetrachloroethene Following Abiotic Versus Biotic Reduction of Hexavalent Chromium

A feasibility evaluation identified chemical reduction and biostimulation as a potential remedy for a plume containing hexavalent chromium (Cr(VI)) and tetrachloroethene (PCE) at an industrial site in southern California. The objectives of this laboratory study were to determine the stoichiometry of calcium polysulfide (CaS_x) reaction with Cr(VI) in the presence of sediment, the effect of CaS_x on the potential for in situ biological reductive dechlorination of PCE, and the potential to reduce Cr(VI) and PCE by addition of only an electron donor. Approximately 1 L of CaS_x solution (containing 50 g S^2-/L) was required per 1000 L of groundwater containing 45 mg/L of Cr(VI) (i.e., 1.8 mol S^2- per mol Cr(VI)). The sediment also exerted a sulfide demand (≥0.38 g S^{2 -} per kg sediment), but at a slower rate than the Cr(VI). In microcosms prepared with lactate, corn syrup, soybean oil, or methanol, but no CaS_x, the Cr(VI) was biologically reduced in the treatments with lactate and corn syrup, but much more slowly than with CaS_x. Even after 20 months of incubation, no significant reductive dechlorination of PCE occurred in any of the microcosms, including those in which the Cr(VI) was removed with CaS_x. Bioaugmentation was tested with the microcosms that received lactate and corn syrup (following 20 months of incubation), using an enrichment culture that actively dechlorinates trichloroethene. PCE dechlorination began within 1 month in the lactate-only treatment; in the corn syrup-amended treatment, PCE dechlorination occurred in only one of the three bottles. However, no PCE dechlorination occurred following bioaugmentation of the lactate and corn syrup microcosms that were initially treated with CaS_x, indicating that CaS_x (and/or its reaction products) exerted a negative impact on the chlororespiring microbes. This outcome highlights the need to evaluate sites on a case-by-case basis when in situ chemical treatment is applied prior to microbial reductive dechlorination.     

Evaluation of Three Electron-Donor Permeable Reactive Barrier Materials for Enhanced Reductive Dechlorination of Trichloroethene

Understanding the fate of complex electron-donor materials is important for developing efficient biostimulation strategies to treat ground water contamination by chlorinated ethenes (CEs). The fermentation product distributions and H₂ production of common permeable reactive barrier (PRB) carbon substrates (dairy whey, sodium lactate syrup, and Hydrogen Release Compound [HRC]) were monitored as measures of substrate efficiency in aquifer microcosms spiked with trichloroethene (TCE). In long-term experiments, the fermentation of PRB substrates to slow-degrading organic acids maintained low H₂ partial pressures (≤ 10^?3.5) that, as previous studies suggest, may give competitive advantage to dechlorinators over hydrogenotrophic methanogens. Whey-amended and lactate-amended microcosms exhibited faster complete dechlorination and, according to organic acid carbon flow, higher rates of fermentation to acetate. In HRC-amended microcosms, propionate appeared to serve as a carbon sink that prolonged dechlorination. Upon complete dechlorination, whey microcosms contained the highest percentage of organic acid carbon. Native Dehalococcoides populations increased by 3 orders of magnitude (per g sediment) in whey-amended microcosms. Whey's efficiency improved in microcosms prepared with aquifer sediment and water from within a downgradient whey PRB. Results suggested whey loading values of 0.2 kg/m³ may be appropriate under sufficiently reducing conditions to efficiently stimulate hydrogenotrophic and potentially actetotrophic dechlorinating populations. Renewal of whey PRBs may, however, be required. Implications for further long-term study of cost-efficiencies are discussed.     

Microbial Reductive Dechlorination of Weathered and Exogenous Co-planar Polychlorinated Biphenyls (PCBs) in an Anaerobic Sediment of Venice Lagoon

The occurrence of reductive dechlorination processes towards pre-existing PCBs and five exogenous coplanar PCBs were investigated in a contaminated sediment of Porto Marghera (Venice Lagoon, Italy) suspended, under strictly anaerobic conditions, in water collected from the same site. PCB dechlorination started after five months of incubation, when sulfate initially occurring in the microcosms was completely depleted and methanogenesis was in progress. It was ascribed to sulfate-reducing bacteria. Several pre-existing hexa-, penta- and tetra-chlorinated biphenyls were slowly bioconverted into tri- and di-, ortho-substituted PCBs from the 5th to the 16th month of experiment. Spiked coplanar PCBs, i.e., 3,3′,4,4′-tetrachlorobiphenyl, 3,3′,4,4′,5- and 2,3′,4,4′,5-pentachlorobiphenyls, 3,3′,4,4′,5,5′- and 2,3,3′,4,4′,5-hexachlorobiphenyls, were extensively transformed (by about 90%) into lower chlorinated congeners, such as 3,3′,5,5′-/2,3′,4,4′-tetrachlorobiphenyl, 3,3′,5-, 2,4,4′-, 2,3′,4- and 2,3′,5-trichlorobiphenyl, 3,4-/3,4′- and 3,3′-dichlorobiphenyl and 2-chlorobiphenyl. The reductive dechlorination of spiked PCBs did not influence significantly the biotransformation rate and extent of pre-existing PCBs.     

Biological Reduction of Trichloroethene Supported by Fe(0)

An anaerobic culture reductively transformed trichloroethene (TCE) in an aqueous medium containing elemental iron as the sole electron source. The TCE disappearance rate was enhanced and the product distribution was markedly altered when the culture was present. In abiotic samples containing Fe(0) but no culture, 11 µmol TCE (equivalent to an aqueous concentration of 260 µM) disappeared over a period of 39 days, with ethene and ethane as the major reduction products. Small amounts of cis-dichloroethene (cis-DCE), 1,1-DCE, and vinyl chloride (VC) also were detected. When the culture was incubated with TCE and Fe(0), the same amount of TCE was transformed in less than 2 weeks. The major products after 39 days were VC, ethene, and ethane. VC accounted for 65% of the initial TCE and appeared to be reduced further to ethene at slow rates. The significant VC production in the culture-amended samples indicates that most TCE was transformed microbially rather than chemically. The data indicate that abiotic and biological reduction of chlorinated ethenes can be coupled to enhance treatment efficiency. The results also suggest that microbial dechlorination within and downgradient from iron walls is potentially important for evaluating the long-term performance of permeable iron barriers.     

Dechlorination of PCE by Mixed Methanogenic Cultures Using Hollow-Fiber Membranes

The study investigated the use of hollow-fiber membranes for hydrogen (H₂) delivery to support the biological reductive dechlorination of tetrachloroethene (PCE) Two experiments were performed in which H₂ was supplied through membranes placed in stirred batch reactors containing a mixed methanogenic/dechlorinating culture and PCE (≤10?µM. Reductive dechlorination of PCE to cis-dichloroethene was sustained in the reactors receiving H₂ (1% H₂ and 50% H₂), while negligible dechlorination was observed in control reactors (100% N₂). The 1%-H₂-fed reactor outperformed the 50%-H₂-fed reactor in the first experiment. However, the dechlorinating performance in the two reactors was similar in the second experiment. Despite relatively high H₂ concentrations (4.6 to 178?µM) that led to H₂ consumption (and CH₄ production) by methanogens, dechlorination was effectively maintained for the duration of the experiments (35 to 62 days). The results of this study are significant in that dechlorination was sustained in a minimal medium by membrane-delivered H₂. Dechlorination was also maintained at aqueous H₂ concentrations that exceeded the thermodynamic thresholds for not only dechlorination (<0.1 to 2?nM, but also methanogenesis (～10?n_M) and homoacetogenesis (94 to 400?n_M. The results of these experiments suggest that membranes are a promising H₂ delivery technology for stimulating the bioremediation of chlorinated ethene-contaminated aquifers.     

Fe0/厌氧微生物联合体系降解硝基苯的研究

利用Fe0/厌氧微生物联合体系降解硝基苯(NB), 结果显示, Fe0与厌氧微生物之间存在明显的协同效应, 硝基苯的降解效果随零价铁投加量的增加而提高;最佳pH值为5.0~6.0;添加少量共代谢初级基质(葡萄糖), 可以大幅度提高硝基苯的降解;较高浓度铁离子对硝基苯的降解表现出一定的抑制作用, 添加0.5 mg/L的Fe3+或Fe2+可以加快硝基苯的降解。硝基苯降解的主要产物为苯胺, 降解过程遵循一级动力学模型, 一级反应速率常数k值随硝基苯浓度的提高而降低。