乳白耙齿菌F17好氧降解四溴双酚A的特性及机理研究

[目的] 为了探究乳白耙齿菌F17（Irpex lacteus F17）降解溴代阻燃剂的可能性,研究了该菌好氧降解四溴双酚A （Tetrabromobisphenol A,TBBPA）的特性以及影响降解的因素,并结合降解产物的分析,推测其降解途径。[方法] 采用高效液相色谱法测定TBBPA的浓度,并通过气相色谱-质谱联用仪分析降解过程的中间产物。[结果] I.lacteus F17可以通过共代谢的方式好氧降解TBBPA,最适共代谢基质是葡萄糖。在葡萄糖浓度为8 g/L、菌悬液接种量为5%、pH 5.0的优化条件下,当TBBPA初始浓度为20 mg/L时降解率可达85.5%,脱溴率为14.6%。对降解过程中锰过氧化物酶的研究发现TBBPA的降解率受到该酶活性的影响。通过气相色谱-质谱联用仪检测到7种中间产物。[结论] I.lacteus F17可以有效降解四溴双酚A,其降解机理主要包括脱溴、β-断裂、羟基化、去质子和氧化等过程。     

山核桃叶总黄酮抑制TBBPA诱导斑马鱼胚胎发育毒性的研究

四溴双酚A(tetrabromobisphenol A,TBBPA)是新型高关注度污染物之一,对鱼类具有强毒性。山核桃叶中的黄酮类化合物具有抗氧化和抗细胞凋亡等作用。通过硅胶柱洗脱、薄层色谱(TLC)检测对山核桃叶进行提取和纯化,得到黄酮类化合物产物B;设置4个实验组(空白对照,0.5 mg/L TBBPA,1.0 mg/L产物B,0.5 mg/L TBBPA和1.0 mg/L产物B),记录斑马鱼胚胎发育过程中的形态学指标;同时,检测不同处理组中活性氧(reactive oxygen species,ROS)水平和凋亡相关基因的表达情况。结果发现,山核桃叶提取的总黄酮产物B中包含50%的查尔酮、27%的黄酮或者黄酮醇、18%的二氢黄酮或者二氢黄酮醇、5%的异黄酮等22种黄酮类化合物;富含黄酮类化合物的产物B对TBBPA诱导斑马鱼胚胎的卵凝结率、畸形率及32 h血流障碍等指标具有显著降低作用;可显著抑制TBBPA诱导的ROS水平升高;对TBBPA诱导的凋亡基因p53上调和Bcl-2下调均有显著改善作用。实验结果表明山核桃叶总黄酮通过降低ROS水平,抑制凋亡基因的异常表达,发挥抗氧化和抑制凋亡作用。实验结果有助于了解山核桃叶有效成分总黄酮对TBBPA诱导的斑马鱼胚胎毒性的保护作用,为水生态系统和人体健康危害的防治提供新思路。     

多溴二苯醚的环境暴露与生态毒理研究进展

多溴二苯醚(PBDEs)是一类具有生态风险的新型环境有机污染物.作为阻燃剂,PBDEs已经被愈来愈广泛地添加到工业产品中,并因此对大气、水体、沉积物和土壤等环境介质及相关生态系统产生日益广泛的污染.鉴于这一环境新问题的产生,本文基于有限的资料,初步探讨了PBDEs的人为来源和环境暴露途径,大致给出了PBDEs在不同生物和人体不同组织器官中可能的存在及含量水平;在扼要介绍其基本性质的基础上,从甲状腺、神经系统和生殖发育毒性等三个方面分析了PBDEs对动物和人体可能产生的毒性效应与生态影响,以及PBDEs在生态系统中可能具有的生物积累和生物放大风险;并对今后研究PBDEs的环境暴露与生态效应以及人体健康影响等方面的工作重点进行了展望.     

溴代阻燃剂微生物降解的研究进展

溴代阻燃剂以其优异的阻燃性能而在工业生产和日常生活中大量使用。这些外源性的化学物质因其蓄积性、持久性、生物毒性而对人类健康和生态环境造成威胁。微生物特有的降解代谢能力为溴代阻燃剂污染治理带来了希望。但是,目前有关溴代阻燃剂的微生物降解研究仍然很少。综述了国内外在溴代阻燃剂微生物降解方面的最新研究动态,包括厌氧、好氧和基团化等生物降解方式。在此基础上,提出目前溴代阻燃剂微生物降解研究中存在的主要问题和重点研究方向,并对其在溴代阻燃剂污染治理方面的应用潜力进行展望。     

溴化马来松香的合成、表征及其阻燃性的研究

本文以马来松香与液溴加成,合成了溴化马来松香;研究了反应温度、原料的摩尔比、反应时间等因素对反应过程及产物性能的影响.用FT-IR、1H NMR和TG对溴化马来松香进行了表征.结果表明,当反应温度为-2～6℃,反应时间5～6 h,反应物的当量配比约为1.2时,收率可达87.9%.溴化马来松香具有较好的阻燃性,同时也拓宽了松香的应用范围,提高了松香的附加值.     

细菌降解双酚类化合物的分子机制研究进展

双酚类化合物(bisphenols,BPs)作为工业原料及药物和个人护理品的重要成分之一,在自然界中广泛分布。BPs作为类雌性激素造成的生态风险已成为全球备受关注的环境问题之一。研究人员使用不同方式分离得到菌株或菌群,尝试对BPs进行无害化降解,目前已取得了一些重要的研究进展。本文对近年来细菌降解BPs的相关研究进行了系统梳理,重点关注以双酚A(bisphenol A,BPA)为典型BPs的细菌降解,总结不同路径中的关键作用基因,讨论相同路径中酶的差异及作用方式,并分析其对BPA的降解效果。本文也简要总结了双酚S(bisphenolS,BPS)和其他BPs的细菌降解研究情况。通过总结讨论现有成果,分析细菌降解BPs中尚需深入研究的内容,探寻未来BPs细菌降解机理及应用的研究方向。     

多溴联苯醚的微生物降解研究进展

多溴联苯醚(Polybrominated diphenyl ethers, PBDEs)作为一种阻燃剂在水体、土壤、沉积物和大气等自然环境中广泛存在。原本认为水溶性低而疏水性高的PBDEs 很难被生物所利用, 但近年研究表明微生物降解是PBDEs 降解的一种重要机制, 部分微生物能参与PBDEs 降解过程。在有氧条件下, 表面活性剂、共代谢基质和初始生长基质等都能影响微生物降解PBDEs; 双加氧酶在PBDEs 降解过程中起着关键作用。电子供体和助溶剂等则在微生物厌氧降解PBDEs 过程中发挥重要作用, 还原脱溴作用是微生物对PBDEs 进行厌氧降解的主要机制。筛选PBDEs 高效降解菌以及进一步揭示其降解机理对于修复PBDEs 污染问题具有重要的理论意义和现实意义。     

有机改性膨润土去除赤潮生物的初步研究

制备了有机膨润土并考察其对中肋骨条藻的去除效果,对中肋骨条藻去除效果是溴化十六烷基三甲铵改性的铁柱撑膨润土>溴化四甲基胺改性的铁柱撑膨润土>铁柱撑膨润土>溴化十六烷基三甲铵改性的钠化膨润土>溴化四甲基胺改性的钠化膨润土>钠化膨润土,有机膨润土对赤潮生物的去除能力随改性剂用量增加而提高,但改性剂用量大于1．0CEC后,增加较少。     

多溴联苯醚的微生物降解机制研究进展

多溴联苯醚(PBDEs)是世界范围内广泛使用的一类溴代阻燃剂,在环境中被频繁检出。因其具有生物积累性、生物毒性和持久性等特点,如今PBDEs已成为全球分布的有机化学毒素。探究PBDEs的降解极为重要,文中从PBDEs微生物降解的角度出发,分别阐释了好氧条件和厌氧条件下细菌降解PBDEs的代谢途径研究进展,并结合原位降解研究推断古菌的降解潜能,比较分析了多种降解途径的特性和综合因素,同时对PBDEs降解微生物未来的研究趋势和PBDEs降解体系设计应用进行了展望。     

南海孔石藻可培养细菌多样性及代谢产物四溴吡咯研究

钙化藻和微生物在造礁石珊瑚幼虫的附着变态过程中扮演着重要角色,对珊瑚礁生态系统的健康具有重要意义。对南海珊瑚礁区钙化藻的共附生细菌进行分离培养,一方面有利于发掘南海珊瑚礁的微生物资源,另一方面有利于进行微生物与珊瑚、钙化藻等珊瑚礁框架生物的相互作用研究。本研究分离获得孔石藻(Porolithon onkodes)表面共附生细菌,基于16S rRNA基因序列进行系统发育分析,并通过高效液相色谱和质谱对分离得到的假交替单胞菌属菌株的代谢产物进行比较分析。从孔石藻分离获得的369株细菌菌株分别隶属于变形菌门(Proteobacteria)、拟杆菌门(Bacteroidetes)和厚壁菌门(Firmicutes)的5纲12目22科47属的97个种级类群。在属级水平上,假交替单胞菌属(Pseudoalteromonas)的菌株数量最多,分属于7个种。在假交替单胞菌属的代谢产物分析中,发现3株在系统发育树中聚为一簇的假交替单胞菌的代谢产物中存在四溴吡咯,而其余没有。研究结果不仅表明了钙化藻中含有丰富的可培养微生物和潜在新物种资源,还首次发现四溴吡咯在南海珊瑚礁区特定类群菌株的代谢产物中的存在,为珊瑚幼虫附着变态的化学信号研究提供基础。     

Thyroid hormonal activity of the flame retardants tetrabromobisphenol A and tetrachlorobisphenol A

The thyroid hormonal-disrupting activity of the flame retardants tetrabromobisphenol A (TBBPA) and tetrachlorobisphenol A (TCBPA) was examined and compared with that of bisphenol A, a typical estrogenic xenobiotic. TBBPA and TCBPA, halogenated derivatives of bisphenol A, markedly inhibited the binding of triiodothyronine (T(3); 1 x 10(-10) M) to thyroid hormone receptor in the concentration range of 1 x 10(-6) to 1 x 10(-4) M, but bisphenol A did not. The thyroid hormonal activity of TBBPA and TCBPA was also examined using rat pituitary cell line GH3 cells, which grow and release growth hormone (GH) depending on thyroid hormone. TBBPA and TCBPA enhanced the proliferation of GH3 cells and stimulated their production of GH in the concentration range of 1 x 10(-6) to 1 x 10(-4) M, while bisphenol A was inactive. TBBPA, TCBPA, and bisphenol A did not show antagonistic action, i.e., these compounds did not inhibit the hormonal activity of T(3) to induce growth and GH production of GH3 cells. TBBPA and TCBPA, as well as bisphenol A, enhanced the proliferation of MtT/E-2 cells, whose growth is estrogen-dependent. These results suggest that TBBPA and TCBPA act as thyroid hormone agonists, as well as estrogens.     

The interaction of the brominated flame retardant: tetrabromobisphenol A with phospholipid membranes

Tetrabromobisphenol A (TBBPA) is one of the most widely used members of the family of brominated flame retardants (BFRs). BFRs, including TBBPA have been shown to be widely distributed within the environment and there is growing evidence of their bio-accumulation within animals and man. Toxicological studies have shown that TBBPA can be harmful to cells by modulating a number of cell signalling processes. In this study, we employed fluorescence spectroscopy and differential scanning calorimetry to investigate the interaction of TBBPA with phospholipid membranes, as this is the most likely route for it to influence membrane-associated cellular processes. TBBPA readily and randomly partitions throughout all regions of the phospholipid bilayer with high efficacy [partition coefficient (Log K(p))=5.7+/-0.7]. A decrease in membrane fluidity in both liquid-crystalline and gel-phase membranes was detected at concentrations of TBBPA as low as 2.5 microM. TBBPA also decreases the phase transition temperature of dipalmitoyl phoshatidylcholine (DPPC) membranes and broadened transition peaks, in a fashion similar to that for cholesterol. TBBPA, however, also prefers to partition into membrane regions not too highly enriched with cholesterol. Our findings therefore suggests that, the toxic effects of TBBPA, may at least in part, be due to its lipid membrane binding/perturbing effects, which in turn, could influence biological processes involving cell membranes.     

Rapid Removal of Tetrabromobisphenol A by Ozonation in Water: Oxidation Products,Reaction Pathways and Toxicity Assessment

Tetrabromobisphenol A (TBBPA) is one of the most widely used brominated flame retardants and has attracted more and more attention. In this work, the parent TBBPA with an initial concentration of 100 mg/L was completely removed after 6 min of ozonation at pH 8.0, and alkaline conditions favored a more rapid removal than acidic and neutral conditions. The presence of typical anions and humic acid did not significantly affect the degradation of TBBPA. The quenching test using isopropanol indicated that direct ozone oxidation played a dominant role during this process. Seventeen reaction intermediates and products were identified using an electrospray time-of-flight mass spectrometer. Notably, the generation of 2,4,6-tribromophenol was first observed in the degradation process of TBBPA. The evolution of reaction products showed that ozonation is an efficient treatment for removal of both TBBPA and intermediates. Sequential transformation of organic bromine to bromide and bromate was confirmed by ion chromatography analysis. Two primary reaction pathways that involve cleavage of central carbon atom and benzene ring cleavage concomitant with debromination were thus proposed and further justified by calculations of frontier electron densities. Furthermore, the total organic carbon data suggested a low mineralization rate, even after the complete removal of TBBPA. Meanwhile, the acute aqueous toxicity of reaction solutions to Photobacterium Phosphoreum and Daphnia magna was rapidly decreased during ozonation. In addition, no obvious difference in the attenuation of TBBPA was found by ozone oxidation using different water matrices, and the effectiveness in natural waters further demonstrates that ozonation can be adopted as a promising technique to treat TBBPA-contaminated waters.     

The interaction of the brominated flame retardant: Tetrabromobisphenol A with phospholipid membranes

Tetrabromobisphenol A (TBBPA) is one of the most widely used members of the family of brominated flame retardants (BFRs). BFRs, including TBBPA have been shown to be widely distributed within the environment and there is growing evidence of their bio-accumulation within animals and man. Toxicological studies have shown that TBBPA can be harmful to cells by modulating a number of cell signalling processes. In this study, we employed fluorescence spectroscopy and differential scanning calorimetry to investigate the interaction of TBBPA with phospholipid membranes, as this is the most likely route for it to influence membrane-associated cellular processes. TBBPA readily and randomly partitions throughout all regions of the phospholipid bilayer with high efficacy {partition coefficient (Log K_p) = 5.7 ± 0.7}. A decrease in membrane fluidity in both liquid-crystalline and gel-phase membranes was detected at concentrations of TBBPA as low as 2.5 μM. TBBPA also decreases the phase transition temperature of dipalmitoyl phoshatidylcholine (DPPC) membranes and broadened transition peaks, in a fashion similar to that for cholesterol. TBBPA, however, also prefers to partition into membrane regions not too highly enriched with cholesterol. Our findings therefore suggests that, the toxic effects of TBBPA, may at least in part, be due to its lipid membrane binding/perturbing effects, which in turn, could influence biological processes involving cell membranes.     

Anaerobic-aerobic process for microbial degradation of tetrabromobisphenol A

Tetrabromobisphenol A (TBBPA) is a flame retardant that is used as an additive during manufacturing of plastic polymers and electronic circuit boards. Little is known about the fate of this compound in the environment. In the current study we investigated biodegradation of TBBPA, as well as 2,4,6-tribromophenol (TBP), in slurry of anaerobic sediment from a wet ephemeral desert stream bed contaminated with chemical industry waste. Anaerobic incubation of the sediment with TBBPA and peptone-tryptone-glucose-yeast extract medium resulted in a 80% decrease in the TBBPA concentration and accumulation of a single metabolite. This metabolite was identified by gas chromatography-mass spectrometry (GC-MS) as nonbrominated bisphenol A (BPA). On the other hand, TBP was reductively dehalogenated to phenol, which was further metabolized under anaerobic conditions. BPA persisted in the anaerobic slurry but was degraded aerobically. A gram-negative bacterium (strain WH1) was isolated from the contaminated soil, and under aerobic conditions this organism could use BPA as a sole carbon and energy source. During degradation of BPA two metabolites were detected in the culture medium, and these metabolites were identified by GC-MS and high-performance liquid chromatography as 4-hydroxybenzoic acid and 4-hydroxyacetophenone. Both of those compounds were utilized by WH1 as carbon and energy sources. Our findings demonstrate that it may be possible to use a sequential anaerobic-aerobic process to completely degrade TBBPA in contaminated soils.     

Anaerobic-Aerobic Process for Microbial Degradation of Tetrabromobisphenol A

Tetrabromobisphenol A (TBBPA) is a flame retardant that is used as an additive during manufacturing of plastic polymers and electronic circuit boards. Little is known about the fate of this compound in the environment. In the current study we investigated biodegradation of TBBPA, as well as 2,4,6-tribromophenol (TBP), in slurry of anaerobic sediment from a wet ephemeral desert stream bed contaminated with chemical industry waste. Anaerobic incubation of the sediment with TBBPA and peptone-tryptone-glucose-yeast extract medium resulted in a 80% decrease in the TBBPA concentration and accumulation of a single metabolite. This metabolite was identified by gas chromatography-mass spectrometry (GC-MS) as nonbrominated bisphenol A (BPA). On the other hand, TBP was reductively dehalogenated to phenol, which was further metabolized under anaerobic conditions. BPA persisted in the anaerobic slurry but was degraded aerobically. A gram-negative bacterium (strain WH1) was isolated from the contaminated soil, and under aerobic conditions this organism could use BPA as a sole carbon and energy source. During degradation of BPA two metabolites were detected in the culture medium, and these metabolites were identified by GC-MS and high-performance liquid chromatography as 4-hydroxybenzoic acid and 4-hydroxyacetophenone. Both of those compounds were utilized by WH1 as carbon and energy sources. Our findings demonstrate that it may be possible to use a sequential anaerobic-aerobic process to completely degrade TBBPA in contaminated soils.     

Determination of tetrabromobisphenol A in human serum by liquid chromatography-electrospray ionization tandem mass spectrometry

A method for the determination of tetrabromobisphenol A (TBBPA) in human serum utilizing solid-phase extractions (SPEs) and liquid chromatography (LC) with electrospray ionization tandem MS (MS/MS) has been developed. After purification and concentration of TBBPA using consecutive SPEs on reversed-phase and normal-phase cartridges, the serum sample was subjected to LC. TBBPA was separated on a C18 reversed-phase column by gradient elution with a mixture of water, methanol, and acetonitrile as the mobile phase, and then detected with electrospray ionization MS/MS in negative ion mode. 13C12-TBBPA was suitable as an internal standard for the reproducible determination of TBBPA in human serum samples (5 g). The method has been validated in TBBPA concentration range of 5-100 pg per g serum, and the recoveries in the concentration range were higher than 83.3%. The repeatabilities of the proposed method of non-spiked control serum (6.3 pg per g serum) and spiked serum (added 5-100 pg per g serum) were within 10.0% as relative standard deviations. The limit of quantification (LOQ) for TBBPA was 4.1 pg per g serum, which was corresponded to 0.63 fmol on column.     

Biodegradation of tetrabromobisphenol A by oxidases in basidiomycetous fungi and estrogenic activity of the biotransformation products

Tetrabromobisphenol A (TBBPA) degradation was investigated using white rot fungi and their oxidative enzymes. Strains of the Trametes, Pleurotus, Bjerkandera and Dichomitus genera eliminated almost 1 mM TBBPA within 4 days. Laccase, whose role in TBBPA degradation was demonstrated in fungal cultures, was applied to TBBPA degradation alone and in combination with cellobiose dehydrogenase from Sclerotium rolfsii. Purified laccase from Trametes versicolor degraded approximately 2 mM TBBPA within 5 h, while the addition of cellobiose dehydrogenase increased the degradation rate to almost 2.5 mM within 3 h. Laccase was used to prepare TBBPA metabolites 2,6-dibromo-4-(2-hydroxypropane-2-yl) phenol (1), 2,6-dibromo-4-(2-methoxypropane-2-yl) phenol (2) and 1-(3,5-dibromo-4-hydroxyphen-1-yl)-2,2',6,6'-tetrabromo-4,4'-isopropylidene diphenol (3). As compounds 1 and 3 were identical to the TBBPA metabolites prepared by using rat and human liver fractions (Zalko et al., 2006), laccase can provide a simple means of preparing these metabolites for toxicity studies. Products 1 and 2 exhibited estrogenic effects, unlike TBBPA, but lower cell toxicity.     

Anti-thyroid hormonal activity of tetrabromobisphenol A, a flame retardant, and related compounds: Affinity to the mammalian thyroid hormone receptor, and effect on tadpole metamorphosis

The thyroid hormone-disrupting activity of tetrabromobisphenol A (TBBPA), a flame retardant, and related compounds was examined. TBBPA, tetrachlorobisphenol A (TCBPA), tetramethylbisphenol A (TMBPA) and 3,3'-dimethylbisphenol A (DMBPA) markedly inhibited the binding of triiodothyronine (T3; 1 x 10(-10) M) to thyroid hormone receptor in the concentration range of 1 x 10(-7)-1 x 10(-4) M, while bisphenol A and 2,2-diphenylpropane were inactive. TBBPA, TCBPA, TMBPA and DMBPA did not exhibit thyroid hormonal activity in a thyroid hormone-responsive reporter assay using a Chinese hamster ovary cell line (CHO-K1) transfected with thyroid hormone receptor alpha1 or beta1, but TBBPA and TCBPA showed significant anti-thyroid hormone effects on the activity of T3 (1 x 10(-8) M) in the concentration range of 3 x 10(-6) - 5 x 10(-5) M. The thyroid hormone-disrupting activity of TBBPA was also examined in terms of the effect on amphibian metamorphosis stimulated by thyroid hormone. TBBPA in the concentration range of 1 x 10(-8) to 1 x 10(-6) M showed suppressive action on T3 (5 x 10(-8) M)-enhancement of Rana rugosa tadpole tail shortening. These facts suggest that TBBPA, TCBPA, TMBPA and DMBPA can act as thyroid hormone-disrupting agents.     

Characterization of an enrichment culture debrominating tetrabromobisphenol A and optimization of its activity under anaerobic conditions

Aim: To study the effects of incubation conditions on the microbial community structure and activity of a TBBPA‐debrominating enrichment culture composed of bacterial and archaeal species. Methods and Results: The effects of the methanogen inhibitor 2‐bromoethanesulfonate (BES), of the antibiotic ampicillin, of substrate (tetrabromobisphenol A, TBBPA) omission and availability of different electron donors on microbial community structure and activity were examined under anaerobic conditions. Debromination of TBBPA was blocked in the presence of ampicillin, while long‐term incubation with BES resulted in delayed debromination activity. The results suggest that the bacterial species responsible for the debromination of TBBPA, while archaeal species involved in electron donor metabolism. The enrichment culture lost its debromination activity after cultivation for 9 months without TBBPA, concomitantly with the disappearance of two DNA bands in a denaturing gradient gel electrophoresis (DGGE) analysis of 16S rRNA gene fragments corresponding to Pelobacter carbinolicus and Sphaerochaeta sp. TQ1 that were present in the original culture. When butyrate was used as an electron donor, TBBPA debromination activity was attenuated. When acetate was used as the electron donor, no debromination was observed and in addition, there was a decrease in the abundance of the mcrA gene. Conclusions: The results indicate that to maintain a high rate of TBBPA debromination activity, it is essential to preserve the microbial community structure (bacterial and archaeal members) of this culture and supply an electron donor that produces high amounts of hydrogen when fermented. Significance and Impact of the Study: The study provides important information for the management of cultures to be used in bioremediation of TBBPA contaminated sites.