一株菲降解细菌的分离及其特性

利用平板熏蒸和双氧加合酶活性测定的方法 ,从大庆油田水体中筛选出 1株对菲具有降解活性的细菌 ,Phx1.在LB培养基中 ,能将浓度为 10 0 μg/mL的菲在 2 4h内完全降解 .与无机盐培养基相比 ,LB和牛肉膏蛋白胨培养基能促进此菌对菲的降解 ,尤其能促进对中间产物的进一步转化 .经质粒消除证明 ,Phx1具有 2个质粒 ,且较大的 1个对菲的降解是必须的 .通过对其形态、生理生化特性 ,16SrDNA序列分析 ,鉴定Phx1为土壤杆菌属细菌 ,其 16SrDNA序列与Agrobacterium tumefaciensUP 3、Agrobacterium albertimagni的相似性最高 ,分别达到 97 8%和 96 6 % ,但对向日葵的致瘤实验为阴性 .     

采用固定化技术处理土壤中菲、芘污染物

采用动胶杆菌(Zoogloea sp.)固定化技术包埋来降解土壤中菲、芘污染物,在不同接种量、不同系列浓度下,对降解效果进行了测定.结果表明:5%的接种量最为合理,在168h时,固定化细菌对菲、芘的降解率达84.89%和76.94%;而在相同条件下土著菌降解仅达到27.85%和19.65%,因此动胶杆菌(Zoogloea sp.)具有更好的降解能力.另外,还用电镜观察研究了动胶杆菌在载体中的分布形态,表明了固定化细菌降解土壤中菲、芘具有较高的优势.     

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.     

Martelella sp. AD-3强化活性污泥耐盐降解菲效能

基于嗜盐菌Martelella sp. AD-3优配市政活性污泥搭建耐盐高效降解菲的生物反应器,在3.0 %盐度,进水菲浓度20mg/L的运行条件下,菲去除率高达97 %.批次实验证明,盐度为3.0 %,pH值为7.5~8.5,底物浓度为20~200mg/L是菲降解的最佳环境条件,此时污泥比活性高于1.0mg/(gVSS×h).各共存底物的受试浓度下,酵母提取物和苯酚促进菲降解,镉和氰化物则抑制该过程,乙酸钠、铜、铬对该过程没有明显影响.16S rRNA基因高通量测序结果表明AD-3菌在反应器内具有长期稳定性,其相对丰度维持在1.5 %.Shinella、Microbacterium和Brevundimonas是反应器中的优势菌,其相对丰度分别为20.7 %,15.1 %和11.9 %.RT-qPCR结果显示接种AD-3菌后,编码PAHs双加氧酶RHDa的功能基因差异倍数从2.1提升至11.7.     

Degradation of polycyclic aromatic hydrocarbons by Pseudomonas sp. JM2 isolated from active sewage sludge of chemical plant

It is important to screen strains that can decompose polycyclic aromatic hydrocarbons (PAHs) completely and rapidly with good adaptability for bioremediation in a local area. A bacterial strain JM2, which uses phenanthrene as its sole carbon source, was isolated from the active sewage sludge from a chemical plant in Jilin, China and identified as Pseudomonas based on 16S rDNA gene sequence analysis. Although the optimal growth conditions were determined to be pH 6.0 and 37℃, JM2 showed a broad pH and temperature profile. At pH 4.5 and 9.3, JM2 could degrade more than 40% of fluorene and phenanthrene (50 mg/L each) within 4 days. In addition, when the temperature was as low as 4℃, JM2 could degrade up to 24% fluorene and 12% phenanthrene. This showed the potential for JM2 to be applied in bioremediation over winter or in cold regions. Moreover, a nutrient augmentation study showed that adding formate into media could promote PAH degradation, while the supplement of salicylate had an inhibitive effect. Furthermore, in a metabolic pathway study, salicylate, phthalic acid, and 9-fluorenone were detected during the degradation of fluorene or phenanthrene. In conclusion, Pseudomonas sp. JM2 is a high performance strain in the degradation of fluorene and phenanthrene under extreme pH and temperature conditions. It might be useful in the bioremediation of PAHs.     

降解芘的分枝杆菌M11的分离鉴定和降解特性

从多环芳烃污染的土壤中分离到1株能高效降解四环芳烃芘的放线菌M11,经形态观察、生理生化和16S rDNA鉴定,属于分枝杆菌属(Mycobacterium sp.).菌株M11能以菲、蒽、荧蒽和芘为唯一碳源生长,在含芘50、100和200 mg/L的无机盐液体培养基中培养16 d降解率分别达到76.9%、91.8%和79.23%.菌株M11对芘的降解具有较广泛的pH范围,在芘浓度100 mg/L,pH为5～9的液体条件下,均可生长.根据已报道的芘降解菌的双加氧酶同源序列设计引物,PCR扩增出编码双加氧酶大亚基和小亚基的基因片段,序列分析表明与已知降解芘的分枝杆菌的双加氧酶基因具有高度同源性.     

假单胞菌P12对菲的降解

从武汉石油化工厂的活性污泥中,分离到一株可以以菲为唯一碳源和能源的假单胞菌P_(12)该菌经靛蓝产生法测定有双加氧酶,在菲无机盐培养基中生长时,有3-菲酚物质积累,经检测有一条质粒带。用丝裂霉素C消除质粒后的菌落,失去了降解菲的能力,说明P_(12)对菲的降解功能是由质粒控制的。     

真菌对土壤中苯并[a]芘的共代谢降解

研究了模拟生物泥浆法修复多环芳烃污染土壤.选择了几种从石油污染土壤中分离出来的真菌,研究它们对土壤内苯并[a]芘的降解,并研究了土壤内共存底物:菲、芘、邻苯二甲酸对苯并[a]芘降解的影响,及其之间的共代谢过程.结果表明,芘可以促进镰刀菌、毛霉对苯并[a]芘的降解,并认为这是共代谢作用的结果;菲也可以促进镰刀菌对苯并[a]芘的降解,但抑制了毛霉和青霉对苯并[a]芘的降解;而邻苯二甲酸对青霉和毛霉降解苯并[a]芘的过程均有抑制作用.      

一株菲高效降解菌的环境适应性研究

生物降解是多环芳烃从环境中去除的主要途径,菲是一种典型的三环芳烃。本研究考察了一株能高效降解多环芳烃菲的鞘氨醇单胞菌GY2B在含河沙环境及不同盐度的人工海水环境中的生长特性与降解菲的情况。结果表明：河沙的加入对菌株GY2B的生长及其高效降解菲的性能均无明显影响,65 h可将起始浓度为100 mg/L的菲降解99.5%以上;而经过驯化后在添加85%人工海水的条件下该菌也仍可正常生长并高效降解菲,66 h可将起始浓度为100 mg/L的菲几乎完全降解。本研究结果可为菌株GY2B在受多环芳烃污染的河滩、河口及近海海洋环境修复中的应用提供参考依据。     

耐冷腐殖酸吸附态PAHs降解菌筛选及其降解特性

为了寻找自然环境中高效PAHs降解菌,并应用于北方寒冷地区PAHs污染土壤修复,从沈抚灌渠冻融土壤中筛选出一株以腐殖酸(HA)吸附态PAHs为碳源和能源且生长良好的耐冷菌株,结合其生理生化特征和16S rDNA序列比对,鉴定此菌株为假单胞菌(Pseudomonas sp.SDR4),并研究了其对冻融土壤中HA吸附态PAHs的降解性能.结果表明:Pseudomonas sp.SDR4对冻融土壤中Phe、Pyr和Ba P均具有一定的降解能力,42 d降解率分别为73.88%、64.88%和49.39%.加入HA能够提高其对Phe、Pyr和Ba P的降解率,42 d降解率分别提高15.9%、13.8%和8.6%;在降解初期加入HA可显著提高Phe、Pyr和Ba P的降解速率,第1周,降解速率分别提高37.8%、28.4%和39.1%,但在后期促进效果减弱.若要在寒冷地区选择Pseudomonas sp.SDR4作为固定化菌株,可添加HA提高其修复效果.本研究为北方寒冷地区PAHs污染土壤修复提供了新的固定化菌种.     

金属离子对蒽、菲、芘生物降解过程的影响

对驯化筛选的多环芳烃(PAHs)高效降解菌CN4与CN2进行了深入研究,澄清了金属离子对其降解多环芳烃的作用,得出结论:对于CN2、CN4降解菌,Mn2+与常量Ca2+、Mg2+同时存在于降解体系中时,对蒽和菲降解起到明显促进作用。Cu2+和常量元素Ca2+、Mg2+混合对菲降解也起到促进作用,但没有Mn2+的促进作用明显。不加任何常量和微量金属元素离子的体系对蒽、菲、芘的降解作用仅次于含Mn2+的降解体系,加入Cu2+、Ca2+、Mg2+、Fe3+、Zn2+对CN2降解蒽、芘产生抑制作用,加入Ca2+、Mg2+、Fe2+、Zn2+对CN2降解菲产生促进作用。Mn2+主要是在降解过程中发挥作用的,外加不同浓度的Mn2+制备的菌悬液,对芘的降解作用基本没有差别。在制备菌悬液时加入Mn2+,菌体对Mn2+有富集作用,但Mn2+的存在与否并不影响菌体的生长繁殖状况。降解体系中加入Mn2+能够有效促进降解过程的进行,加入0.5 mmol/L的Mn2+对CN4菌株降解芘的促进作用最大,而在降解体系中一次性加入5.0 mmol/L的Mn2+则会对降解产生抑制作用。     

Isolation of marine benzo[a]pyrene-degrading Ochrobactrum sp. BAP5 and proteins characterization

A bacterial strain BAP5 with a relatively high degradation ability of benzo[a]pyrene (BaP) was isolated from marine sediments of Xiamen Western Sea, China and identified as Ochrobactrum sp. according to 16S rRNA gene sequence as well as Biolog microbial identification system. Strain BAP5 could grow in mineral salt medium with 50 mg/L of BaP and degrade about 20% BaP after 30 d of incubation. Ochrobactrum sp. BAP5 was able to utilize other polycyclic aromatic hydrocarbons (PAHs) (such as phenanthrene, pyrene and fluoranthene) as the sole carbon source and energy source, suggesting its potential application in PAHs bioremediation. The profile of total soluble protein from Ochrobactrum sp. BAP5 was also investigated. Some over- and special-expressed proteins of strain BAP5 when incubated with the presence of BaP were detected by two-dimensional polyacrylamide gel electrophoresis, and found to be related with PAHs metabolism, DNA translation, and energy production based on peptide fingerprint analysis through matrix-assisted laser desorption/ionization-time of flight mass spectrometry.     

一株高效降解菲的植物内生细菌筛选及其生长特性

从长期受PAHs污染的植物看麦娘中分离出1株可高效降解菲的内生细菌Pn2,经生理生化特征分析和16S rDNA序列同源性分析,初步鉴定为Naxibacter sp.,研究了菌株Pn2的生长特性及其对菲的降解作用.结果表明,菌株Pn2能以菲为唯一碳源生长,并对菲有良好的降解性能.菲浓度为49.92mg/L时,30℃下150r/min振荡培养72h,菲降解率高达98.78%.接种量和污染强度显著影响Pn2对菲的降解:接种量越大,菲降解率越高;随污染强度升高,菲降解率先增大后减小,最适污染强度为150mg/L.菌株Pn2有较强的环境适应能力.温度为25~37℃、环境pH值为6.0~8.0、盐浓度1%~2%范围内,菌株Pn2生长状况良好.菌株Pn2为好氧生长,通气量越大,菌株生长越旺盛.菌株Pn2对低浓度的氨苄青霉素和氯霉素有抗性.     

两株假单胞菌对蒽菲芘的降解作用

从污染污泥中分离出两株假单胞菌PCN5及PCB2。研究了它们对蒽、菲、芘的降解性能及生长繁殖情况。结果表明，单基质存在下，10h，PCN5对蒽的降解转化率为91．8％，芘为75．6％，菲仅为26．25％；相反PCB2对菲的降解效果最好，蒽最差；混合基质体系中，两菌株都有良好的降解效果，对芘的降解效果较差；130h，PCN5对蒽、菲、芘混合体系中的TOC去除率为38．9％，PCB2对相应体系的去除率为73．7％；混合体系中两株的生长曲张相似，细菌浓度均呈指数增长趋势，PCN5的最大浓度约是原加入量的10000倍，PCB2是原加入量的8000倍。     

Isolation of marine benzo[a]pyrene-degrading Ochrobactrum sp. BAP5 and proteins characterization

A bacterial strain BAP5 with a relatively high degradation ability of benzo[a]pyrene (BaP) was isolated from marine sediments of Xiamen Western Sea, China and identified as Ochrobactrum sp. according to 16S rRNA gene sequence as well as Biolog microbial identification system. Strain BAP5 could grow in mineral salt medium with 50 mg/L of BaP and degrade about 20% BaP after 30 d of incubation. Ochrobactrum sp. BAP5 was able to utilize other polycyclic aromatic hydrocarbons (PAHs) (such as phenanthrene, pyrene and fluoranthene) as the sole carbon source and energy source, suggesting its potential application in PAHs bioremediation. The profile of total soluble protein from Ochrobactrum sp. BAP5 was also investigated. Some over- and special-expressed proteins of strain BAP5 when incubated with the presence of BaP were detected by two-dimensional polyacrylamide gel electrophoresis, and found to be related with PAHs metabolism, DNA translation, and energy production based on peptide fingerprint analysis through matrix-assisted laser desorption/ionization-time of flight mass spectrometry.     

青顶拟多孔菌对单一和复合多环芳烃的降解特性

利用中国东北林区普遍存在白腐菌——青顶拟多孔菌,降解单一和复合多环芳烃,分别测定了菲、蒽、芘于11,22,33d的累积降解率.结果显示,对于单一多环芳烃,该菌种降解能力由强到弱依次为菲>蒽>芘,33d累积降解率依次为96.56%、94.76%和57.53%;对复合多环芳烃降解中,菲和芘的累积降解率分别为99.46%和61.09%.在复合多环芳烃的降解研究中发现,少量蒽的加入,刺激了菌种对菲和芘的降解,使菲和芘的降解率分别提高了2.9%和3.56%.由此提示,在研究降解高环、难降解多环芳烃时,可利用低环多环芳烃对菌种的刺激作用,在体系内形成高、低环多环芳烃的共代谢,以达到更加高效降解多环芳烃的目的.     

优势短杆菌对多环芳烃的降解性能

本研究从长期被焦化废水污染的污泥中分离出一株优势短杆菌研究该菌株及其在无机离子存在下对蒽、菲、芘的降解性能.结果发现该菌株对蒽、菲、芘均有良好的降解效果,Fe³⁺对降解过程有促进作用,反应10h后,蒽、菲、芘转化率分别可达约75%,87%,及62%.总有机碳(TOC)的测定结果表明,该菌株在106h内能分别将反应瓶中加入的蒽、菲、芘所产生的总有机碳去除50%、90%及50%.     

三株菲降解细菌的分离、鉴定及降解特性的研究

从石油污染土壤中分离纯化到 3株能以菲为唯一碳源和能源生长的革兰氏阴性杆菌、好氧、不形成芽孢、端生或侧生单根鞭毛 ,分别命名为菌株ZX4、ZX6和EVA17.据部分片段长度的 16SrDNA序列比对分析和生理生化试验结果 ,确认此 3株菌均为鞘氨醇单胞菌属 (Sphingomonas)细菌 ,菌株ZX4为少动鞘氨醇单胞菌 (S .paucimobilis) ,菌株ZX6为溶芳烃鞘氨醇单胞菌 (S .aromaticivorans) ,菌株EVA17尚不能确定 .分析了各菌株在系统发育中的分类地位 .菌株ZX4在 14d内对含量为 10 0 0mg·L-1的菲降解率可达 98 74%.不同底物实验结果表明 ,菌株EVA17可能同时拥有两条菲降解途径 .     

生物炭固定化多环芳烃高效降解菌剂的制备及稳定性

高效多环芳烃降解微生物在污染环境中存活并保持一定生物量是实现生物强化修复的前提.本研究通过优选生物炭,优化生物炭固定化多环芳烃高效降解菌Martelella sp.AD-3的制备条件以及评估生物炭固定化菌剂的稳定性,期望获得具有应用前景的生物材料.结果显示,稻壳生物炭比表面积及孔隙大、Zeta电位高、固定化菌剂去除效果好,选择其作为固定化AD-3菌的载体.电镜观察及固定化菌剂对菲的去除率表明,固定化培养基为3% LB,接种量为2.9×10⁸ CFU·mL^-1,固定2 d时,稻壳固定化菌剂负载AD-3量最多,对菲的去除速率可达8.08 mg·L^-1·h^-1.室温保存21 d后,稻壳生物炭固定化菌剂对菲的去除速率仍达到4.46 mg·L^-1·h^-1,表明稻壳生物炭固定化AD-3菌不仅保持了菌对菲的高效降解能力,而且延长了降解微生物的保存时间,这为多环芳烃污染土壤修复提供了良好的生物修复功能材料.     

Photocatalytic degradation of phenanthrene on soil surfaces in the presence of nanometer anatase TiO2 under UV-light

The effect of nanometer anatase TiO₂ was investigated on the photocatalytic degradation of phenanthrene on soil surfaces under a variety of conditions. After being spiked with phenanthrene, soil samples loaded with different amounts of TiO₂ (0 wt.%, 1 wt.%, 2 wt.%, 3 wt.%, and 4 wt.%) were exposed to UV-light irradiation for 25 hr. The results indicated that the photocatalytic degradation of phenanthrene followed the pseudo first-order kinetics. TiO₂ significantly accelerated the degradation of phenanthrene with the half-life reduced from 45.90 to 31.36 hr for TiO₂ loading of 0 wt.% and 4 wt.%, respectively. In addition, the effects of H₂O₂, light intensity and humic acid on the degradation of phenanthrene were investigated. The degradation of phenanthrene increased with the concentration of H₂O₂, light intensity and the concentration of humic acids. It has been demonstrated that the photocatalytic method in the presence of nanometer anatase TiO₂ was a very promising technology for the treatments of soil polluted with organic substances in the future.