细菌还原U(Ⅵ)分子生物学机理的研究进展

铀(U)污染对生态环境和人类健康的潜在危害受到越来越多的关注.U(Ⅵ)还原细菌可将U(Ⅵ)还原至U(Ⅳ),从而降低铀在水中的溶解性和移动性,达到污染修复的目的.目前发现的U(Ⅵ)还原细菌主要包括但不限于铁还原菌和硫酸盐还原菌.本文综述了细菌还原U(Ⅵ)的分子生物学机理,重点阐述了U(Ⅵ)还原细菌的胞外电子转移方式,包括希瓦氏菌的金属还原方式、土杆菌的孔蛋白介导方式和微生物纳米线方式.竞争性电子受体和共存离子对细菌还原U(Ⅵ)有重要影响.目前细菌还原U(Ⅵ)过程中胞外电子转移的机理仍需更多实证,土杆菌利用微生物纳米线和细胞色素协作调控电子转移的机制尚不明确.今后可将研究聚焦于细菌还原U(Ⅵ)机理的验证和完善,并开发和优化基于微生物还原的铀污染修复技术,进而提高铀污染生物修复效率和稳定性.     

灭活与非灭活条件下植物乳杆菌去除U(VI)的机理

在不同时间,pH值和生物量浓度条件下,进行了灭活与非灭活植物乳杆菌去除水中铀的对比试验,探讨了二者去除水中铀的机理,通过SEM-EDS、FTIR、XPS及XRD分析了铀与菌体表面的微观作用机理以及菌体表面沉积物的特征.结果表明：植物乳杆菌经灭活后,其吸附铀的能力得到显著的提高,当U（VI）初始浓度为10mg/L、pH值为6.0、37℃条件下,120min内灭活菌体对U（VI）的去除率为94.7%,而活菌体的去除率为88.9%.灭活菌体具有更高的铀吸附容量,在生物量浓度为0.06~0.24mg/L,pH值（3.0~7.0）条件下,灭活菌体与活菌体的U（VI）累积容量比W均大于1.SEM-EDS、FTIR分析结果表明,活细胞和灭活细胞都可通过细胞表面的羟基、酰基及羧基等官能团吸附、配位络合U（VI）.XRD分析表明,活菌体可生物磷酸矿化水中的U（VI）.活菌体的XRD谱图在2θ（18.023,25.492,27.343,40.813°处）有4个明显的磷酸铀酰晶体峰,而灭活菌体的XRD谱图显示为非晶态.XPS结果表明,活菌体可生物还原U（VI）.活菌体能谱图中U4f7/2和U4f5/2轨道出现了结合能为380.20eV和390.65eV的U（VI）分裂峰,而灭活菌体的能谱图中没有出现U（IV）的分裂峰.     

嗜根寡养单胞菌DSM14405T对高浓度Cr(Ⅵ)的抗性及其还原特性分析

为了寻找植物-微生物联合治理Cr（VI）污染的合适微生物,分析了一株植物根际促生菌——嗜根寡养单胞菌DSM 14405^T（Stenotrophomonas rhizophila DSM 14405^T）对不同浓度Cr（VI）的还原能力、Cr（VI）还原最适培养条件、对连续投加Cr（VI）的还原能力及高浓度Cr（VI）处理不同时间的转录组变化.结果表明,嗜根寡养单胞菌DSM 14405^T在24 h内对10、50、200和500 mg·L^-1 Cr（VI）的还原率分别为100%、92.4%、26.2%和16.5%,且在30℃、pH=7.5、转速180 r·min^-1条件下培养能最经济、高效地还原Cr（VI）.该菌能在64 h内完全还原2次所投加的50 mg·L^-1的Cr（VI）,第3次投加后仅能还原55.2%左右.比较转录组学分析可得,200 mg·L^-1Cr（VI）作用下,嗜根寡养单胞菌DSM 14405^T细胞表现出显著的抗性.具体表现为：大幅度上调胞内消除有毒自由基的相关基因、DNA修复相关基因以防止DNA因自由基受损,同时下调细胞组分及蛋白代谢等与细胞抗性相关性低的通路.并且,随着Cr（VI）的还原和细胞对环境的适应,Cr（VI）抗性相关基因及通路的调节在长时暴露中明显少于短时暴露.本研究为Cr（VI）污染的原位植物-微生物联合治理提供了可选菌株和理论依据.     

沉积物电缆细菌研究进展

目前,在沉积物中发现一种丝状多细胞微生物-电缆细菌,该菌属于变形菌门的脱硫球茎菌科（Desulfobulbaceae）.电缆细菌通过远距离电子传输将沉积物表面的氧还原与深处缺氧层的硫化物氧化电耦合进行完整的氧化还原反应,调节氧分子和硫化物迁移转化.电缆细菌有3种连接方式,表层存在脊段结构进行电子运输,并在富含硫酸盐和季节性周期变化的自然环境中广泛存在.这种新型的多细胞合作方式对沉积物的生物地球化学循环产生显著影响,促进沉积物硫化区铁锰的溶解,并抑制硫和磷等元素的释放.此外,电缆细菌参与硫化物和烃类污染物的降解,为污染沉积物生物修复领域提供新的研究方向.本文对电缆细菌的发现、生理代谢、栖息环境以及生物地球化学循环的影响等研究进行了综述.     

地下水铀污染的原位微生物还原与固定:在美国能源部田纳西橡树岭放射物污染现场的试验

总结了美国斯坦福大学和橡树岭国家实验室等在美国能源部田纳西州橡树岭综合试验基地进行的铀污染原位微生物修复阶段性试验结果.本试验利用微生物以乙醇为电子供体还原地下水和沉积物中的六价铀为不溶解的四价铀,使之原位固定化.随后通过加入溶解氧和硝酸盐来试验微生物还原后的地下水层中还原固定态铀的稳定性.通过预处理和长期间隔注入乙醇...     

菌株Enterobacter sp. L6异化Fe(III)还原及Cr(VI)还原的性质分析

Cr（VI）是一种毒性极强的重金属,利用微生物还原Cr（VI）为Cr（III）是解决Cr（VI）污染的一条有效途径。菌株Enterobacter sp. L6是一株分离自海洋沉积物中的异化铁还原细菌。接种时细胞密度A₆₀₀为（0.25±0.03）,培养12 h,A₆₀₀达到（1.04±0.05）,累积产生Fe（II）浓度为（0.80±0.03）mmol/L;随着培养时间的延长,细胞密度A₆₀₀和累积产生Fe（II）浓度开始下降;培养36 h时,细胞密度A₆₀₀为（0.81±0.04）,累积Fe（II）浓度（0.63±0.01）mmol/L。在厌氧培养过程中,菌株L6细胞生长与异化还原Fe（III）性质存在明显的偶联关系。利用菌株L6的异化铁还原性质还原Cr（VI）的实验结果表明,在Cr（VI）浓度0～24 mg/L范围内,异化铁还原细菌L6都能进行细胞生长并还原Cr（VI）。Cr（VI）浓度为4、8和12 mg/L时,菌株L6对Cr（VI）还原率可达到100%,当Cr（VI）浓度为16 mg/L时,Cr（VI）还原率是参比[未添加Fe（III）]的2.11倍。Cr（VI）浓度为20、24 mg/L时,仍能够还原Cr（VI）。以Fe（III）为电子受体的异化铁还原细菌能明显提高Cr（VI）还原率,这为利用微生物修复Cr（VI）污染提供实验数据支持。     

电场强化博落回修复铀镉复合污染土壤机理

采用盆栽试验,研究了在0.3V/cm的外加直流电场作用下,博落回的生物量、富集铀（U）的性能和抗氧化酶活性,以及其根际土壤中有机酸含量、U和镉（Cd）的结合形态、植物根部U的价态、微生物群落结构的变化等.结果表明,施加直流电场后,博落回总生物量升高了15.33%~29.88%,其中电场+铀污染（DC+U）和电场+镉污染（DC+Cd）处理组的博落回对U和Cd的富集系数提高了90.84%和93.33%;土壤中草酸、酒石酸、琥珀酸、苹果酸和乳酸含量分别增加了18.36%~45.31%、58.62%~503.22%、15.71%~118.99%、12.34%~123.27%和25.97%~36.05%;过氧化物酶（POD）和谷胱甘肽过氧化物酶（GSH-PX）的活性分别提高了13.63%~34.82%和9.70%~28.64%;根际土壤中植物可利用态U、Cd所占比例显著增大;博落回根部的大部分U由稳定的U（IV）变成了更容易从地下部分向地上部分转移的U（VI）;酸杆菌门（Acidobacteria）等细菌菌门和子囊菌门（Ascomycota）等真菌菌门比例升高,这些微生物通过提高酶活性增强了博落回对U、Cd的耐受性和富集作用.     

电场强化博落回修复铀镉复合污染土壤机理

采用盆栽试验,研究了在0.3V/cm的外加直流电场作用下,博落回的生物量、富集铀（U）的性能和抗氧化酶活性,以及其根际土壤中有机酸含量、U和镉（Cd）的结合形态、植物根部U的价态、微生物群落结构的变化等.结果表明,施加直流电场后,博落回总生物量升高了15.33%~29.88%,其中电场+铀污染（DC+U）和电场+镉污染（DC+Cd）处理组的博落回对U和Cd的富集系数提高了90.84%和93.33%;土壤中草酸、酒石酸、琥珀酸、苹果酸和乳酸含量分别增加了18.36%~45.31%、58.62%~503.22%、15.71%~118.99%、12.34%~123.27%和25.97%~36.05%;过氧化物酶（POD）和谷胱甘肽过氧化物酶（GSH-PX）的活性分别提高了13.63%~34.82%和9.70%~28.64%;根际土壤中植物可利用态U、Cd所占比例显著增大;博落回根部的大部分U由稳定的U（IV）变成了更容易从地下部分向地上部分转移的U（VI）;酸杆菌门（Acidobacteria）等细菌菌门和子囊菌门（Ascomycota）等真菌菌门比例升高,这些微生物通过提高酶活性增强了博落回对U、Cd的耐受性和富集作用.     

Shewanella oneidensis MR-1对Cr(VI)的还原及其影响因素

在实验室纯培养条件下,探讨厌氧体系中Shewanella oneidensis MR-1对Cr（VI）的还原能力,采用扫描电镜（SEM）-能谱（EDS）、X射线光电子能谱（XPS）等方法进行表征.结果表明,S.oneidensis MR-1介导下不同浓度Cr（VI）的生物转化与微生物对铬的耐受特性密切相关,低浓度Cr（VI）对其生长影响不大,高浓度时细菌生长则受到抑制,进而抑制Cr（VI）的还原率;菌株对Cr（VI）的还原作用随着接种菌悬液量的增加而增强;菌株最适生长pH值为中性,弱碱性环境比酸性环境更有利于菌株对Cr（VI）的还原;增加Fe（Ⅲ）的量会加快Cr（VI）完全还原的速率.通过SEM-EDS和XPS分析,在对Cr（VI）进行处理5d后,菌体表面有Cr（VI）和Cr（Ⅲ）两种形态存在,证实S.oneidensis MR-1在对Cr（VI）进行还原的同时也伴有少量的吸附作用.微生物还原为环境中Cr（VI）的去除以及解毒提供了一种有效的方法.     

低浓度铀污染地下水的生物还原矿化修复研究进展

随着核能资源的开发利用,低浓度铀污染地下水引起的环境问题日益突显,如何高效低耗地处理低浓度铀污染地下水成为当今的研究热点。生物还原和生物矿化是修复低浓度铀污染地下水的2种有效方法,而相关研究主要集中在单一机制上,生物还原耦合生物矿化修复低浓度铀污染地下水的研究还缺少系统报道。文章简述了生物还原和生物矿化的机制、参与微生物以及反应产物的稳定性,论述了生物还原耦合生物矿化修复低浓度铀污染地下水的优势、作用机制,同时从微生物和环境因子2个方面总结了耦合机制修复低浓度铀污染地下水的影响因素,提出生物还原耦合生物矿化修复低浓度铀污染地下水的强化方法,并对该技术的应用前景进行了展望。     

Microbial reduction of uranium(Ⅵ) by Bacillus sp.dwc-2:A macroscopic and spectroscopic study

The microbial reduction of U(VI) by Bacillus sp. dwc-2, isolated from soil in Southwest China,was explored using transmission electron microscopy(TEM), X-ray photoelectron spectroscopy(XPS) and X-ray absorption near edge spectroscopy(XANES). Our studies indicated that approximately 16.0% of U(VI) at an initial concentration of 100 mg/L uranium nitrate could be reduced by Bacillus sp. dwc-2 at pH 8.2 under anaerobic conditions at room temperature.Additionally, natural organic matter(NOM) played an important role in enhancing the bioreduction of U(VI) by Bacillus sp. dwc-2. XPS results demonstrated that the uranium presented mixed valence states(U(VI) and U(IV)) after bioreduction, which was subsequently confirmed by XANES. Furthermore, the TEM and high resolution transmission electron microscopy(HRTEM) analysis suggested that the reduced uranium was bioaccumulated mainly within the cell and as a crystalline structure on the cell wall.These observations implied that the reduction of uranium may have a significant effect on its fate in the soil environment in which these bacterial strains occur.     

Enhanced U(VI) bioreduction by alginate-immobilized uranium-reducing bacteria in the presence of carbon nanotubes and anthraquinone-2,6-disulfonate

Uranium-reducing bacteria were immobilized with sodium alginate, anthraquinone-2, 6-disulfonate (AQDS), and carbon nanotubes (CNTs). The effects of different AQDS-CNTs contents, U(IV) concentrations, and metal ions on U(IV) reduction by immobilized beads were examined. Over 97.5% U(VI) (20 mg/L) was removed in 8 hr when the beads were added to 0.7% AQDS-CNTs, which was higher than that without AQDS-CNTs. This result may be attributed to the enhanced electron transfer by AQDS and CNTs. The reduction of U(VI) occurred at initial U(VI) concentrations of 10 to 100 mg/L and increased with increasing AQDS-CNT content from 0.1% to 1%. The presence of Fe(III), Cu(II) and Mn(II) slightly increased U(VI) reduction, whereas Cr(VI), Ni(II), Pb(II), and Zn(II) significantly inhibited U(VI) reduction. After eight successive incubation-washing cycles or 8 hr of retention time (HRT) for 48 hr of continuous operation, the removal efficiency of uranium was above 90% and 92%, respectively. The results indicate that the AQDS-CNT/AL/cell beads are suitable for the treatment of uranium-containing wastewaters.     

Microbial reduction of uranium (VI) by Bacillus sp. dwc-2: A macroscopic and spectroscopic study

The microbial reduction of U(VI) by Bacillus sp. dwc-2, isolated from soil in Southwest China, was explored using transmission electron microscopy (TEM), X-ray photoelectron spectroscopy (XPS) and X-ray absorption near edge spectroscopy (XANES). Our studies indicated that approximately 16.0% of U(VI) at an initial concentration of 100 mg/L uranium nitrate could be reduced by Bacillus sp. dwc-2 at pH 8.2 under anaerobic conditions at room temperature. Additionally, natural organic matter (NOM) played an important role in enhancing the bioreduction of U(VI) by Bacillus sp. dwc-2. XPS results demonstrated that the uranium presented mixed valence states (U(VI) and U(IV)) after bioreduction, which was subsequently confirmed by XANES. Furthermore, the TEM and high resolution transmission electron microscopy (HRTEM) analysis suggested that the reduced uranium was bioaccumulated mainly within the cell and as a crystalline structure on the cell wall. These observations implied that the reduction of uranium may have a significant effect on its fate in the soil environment in which these bacterial strains occur.     

共存离子对地浸废水中铀生物沉淀过程的影响

采用硫酸盐还原菌生物(还原)沉淀法消除地浸废水中的放射性铀污染,研究了共污染离子Cu(II)、Zn(II)、Fe(III)和SO42-分别对铀生物沉淀过程的影响。序批式实验结果表明,初始Cu(II)浓度低于10mg/L或Zn(II)浓度低于20mg/L时对铀生物沉淀过程影响不大,当Cu(II)浓度超过15mg/L或Zn(II)浓度超过25mg/L时,该过程会因重金属的生物毒性作用受到完全抑制。在含有Fe(III)的氧化环境中,铀生物沉淀过程与Fe(III)还原过程同时进行,但铀沉淀速度相对减慢。初始SO42-浓度低于4000mg/L时对铀生物沉淀过程影响很小,超过5000mg/L时会产生明显的抑制作用,且抑制作用随着SO42-浓度的上升而加强。     

微生物溶解载铀赤铁矿及再固定铀的试验研究

将载铀赤铁矿、核黄素(RF)和Sphingomonas sanxanigenens(S. sanxanigenens)同时投加到培养基中,监测培养过程中溶液的总铁、亚铁以及U(VI)浓度的变化,表征载铀赤铁矿还原性溶解前后固相产物中铁和铀的化学形态与价态,分析温度、共存离子对RF介导S. sanxanigenens还原性溶解载铀赤铁矿及再固定铀的影响.结果表明:S. sanxanigenens能够还原性溶解载铀赤铁矿,从而导致铀的释放;RF能够促进S. sanxanigenens还原性溶解载铀赤铁矿,且RF浓度越高越有利于这种还原性溶解;在30℃下RF能显著促进载铀赤铁矿的还原性溶解,且产物中稳定态铀的比例较高;添加2mmol/L Ca²⁺或CO₃^2-对RF介导S. sanxanigenens还原性溶解载铀赤铁矿具有促进作用;添加2mmol/L PO₄^3-能提高残渣态铀的比例,促进铀的固定;RF能够促进S. sanxanigenens对Fe(III)和U(VI)的还原,且反应过程伴随着次生铁矿物的生成.这些研究结果为提出RF介导S. sanxanigenens释放及再固定铀的新方法奠定了基础.     

奥奈达希瓦氏菌MR-1还原U(VI)的特性及影响因素

探讨了在腐殖质模式物蒽醌?2?磺酸钠(AQS)存在条件下,奥奈达希瓦氏菌MR-1的还原U(VI)特性.结果表明,在厌氧环境下奥奈达希瓦氏菌以AQS为电子穿梭载体,利用电子供体高效还原U(VI).当菌体投加量为1.2×109 个时,其还原铀的效率达95.09%; AQS的浓度低于0.5mmol/L时有利于MR-1菌厌氧还原U(VI),AQS浓度的升高U(VI)的还原明显受到抑制.当U(VI)初始浓度为30.0mg/L时,分别以甲酸盐、乙酸盐和乳酸盐为电子供体,经过7d后其还原率分别达到95.37%、92.41%和95.65%.金属离子(Cu2+、Mn2+、Ca2+)、有毒有机物等对U(VI)还原产生影响.当Ca2+的浓度为2.0mmol/L时,对U(VI)的还原有微弱的促进作用,而当Cu2+和Mn2+浓度为2.0mmol/L时,则存在较强的抑制作用.奥奈达希瓦氏菌也能利用环境中甲苯、三氯乙酸、顺丁烯二酸等有毒物质高效还原U(VI),同时使有毒物质得到降解.扫描电子显微镜(SEM)和电子能谱(EDS)分析结果表明,奥奈达希瓦氏菌菌体中沉积了铀元素.     

Mechanistic insights into sequestration of U(VI) toward magnetic biochar: Batch, XPS and EXAFS techniques

The magnetic iron oxide(Fe_3O_4) nanoparticles stabilized on the biochar were synthesized by fast pyrolysis of Fe(II)-loaded hydrophyte biomass under N_2 conditions. The batch experiments showed that magnetic biochar presented a large removal capacity(54.35 mg/g)at pH 3.0 and 293 K. The reductive co-precipitation of U(VI) to U(IV) by magnetic biochar was demonstrated according to X-ray diffraction, X-ray photoelectron spectroscopy and X-ray absorption near edge structure analysis. According to extended X-ray absorption fine structure analysis, the occurrence of U-Fe and U-U shells indicated that high effective removal of uranium was primarily inner-sphere coordination and then reductive co-precipitation at low pH. These observations provided the further understanding of uranium removal by magnetic materials in environmental remediation.     

Aspergillus tubingensis介导植酸盐水解促进U(VI)-PO43-生物矿化

从广东某铀尾矿库水下沉积物中分离筛选出了一株能水解植酸盐的真菌M5-1,对其菌落形态、ITS序列、最适生长pH值、对铀的耐受性及其水解植酸盐的效果进行了分析,随后对M5-1生物矿化铀过程中pH值、正磷酸盐浓度、铀浓度、铀去除率的变化进行了监测,对矿化产物的主要元素和矿物组成进行了分析.证实了真菌M5-1为Aspergillus tubingensis（MH978623）,其最适生长pH值范围为6~7,对铀（~0.84mmol/L）具有较强的耐受性;Aspergillus tubingensis介导植酸盐水解促进U（VI）-PO₄^3-矿化62d后,铀的去除率达95.2%;Aspergillus tubingensis介导U（VI）-PO₄^3-矿化过程中可能形成了难溶的氢铀云母和变钠铀云母矿物.结果表明,Aspergillus tubingensis能有效水解植酸盐释放可溶性正磷酸盐,从而促进U（VI）-PO₄^3-矿化.研究结果为采用Aspergillus tubingensis介导植酸盐水解原位修复铀污染地表水提供了试验依据.     

Herbicide promotes the conjugative transfer of multi-resistance genes by facilitating cellular contact and plasmid transfer

The global dissemination of antibiotic resistance genes (ARGs), especially via plasmid-mediated horizontal transfer, is becoming a pervasive health threat. While our previous study found that herbicides can accelerate the horizontal gene transfer (HGT) of ARGs in soil bacteria, the underlying mechanisms by which herbicides promote the HGT of ARGs across and within bacterial genera are still unclear. Here, the underlying mechanism associated with herbicide-promoted HGT was analyzed by detecting intracellular reactive oxygen species (ROS) production, extracellular polymeric substance composition, cell membrane integrity and proton motive force combined with genome-wide RNA sequencing. Exposure to herbicides induced a series of the above bacterial responses to promote HGT except for the ROS response, including compact cell-to-cell contact by enhancing pilus-encoded gene expression and decreasing cell surface charge, increasing cell membrane permeability, and enhancing the proton motive force, providing additional power for DNA uptake. This study provides a mechanistic understanding of the risk of bacterial resistance spread promoted by herbicides, which elucidates a new perspective on nonantibiotic agrochemical acceleration of the HGT of ARGs.