红串红球菌TJQ对不同形式硫的脱除研究

红串红球菌TJQ是本实验室筛选的一株专一性脱硫菌,高效液相色谱法（HPLC）分析表明．该菌能选择性地脱除二苯并噻吩（DBT）中的硫,最终代谢产物是2-羟基联苯（2-HBP）。实验结果表明,该菌既可以利用有机硫源又可以利用廉价的无机硫源作为生长硫源,以硫酸钠作为硫源培养的菌株可专一性脱除苯并噻吩（BT）和DBT及其甲基衍生物4,6-二甲基二苯并噻吩（4,6-DMDBT）中的硫。除此之外,它还可以脱除苯硫醚（PS）和硫醇类含硫化合物中的硫。在正十六烷模拟体系中菌株对噻吩（TH）、4,6-DMDBT、DBT等类型的硫的降解效率很高,对BT和PS类硫的降解效率也较好。它可以使实际柴油中总硫含量由554mg／L降到267mg／L,降解率达到51．8%。     

苯并噻吩与二苯并噻吩脱硫酶功能相关性研究

苯并噻吩（BT）专一性降解菌株Gordonia sp. C-6的脱硫途径类似于二苯并噻吩（DBT）的“4S”脱硫途径,能以BT为唯一硫源生长,但不能以DBT为唯一硫源生长.目前,还没有BT专一性脱硫基因的相关报道.本研究将Rhodocossus erythropolis DS-3中DBT脱硫途径的相关基因dszA、dszB、dszC和dszABC分别转入Gordonia sp. C-6中构建工程菌株Gordonia sp. CRA、Gordonia sp. CRB、Gordonia sp. CRC和Gordonia sp. CRABC;其DBT相关脱硫酶活性(以DCW计)分别为76.8 μmol·(g·h)^-1、 51.6 μmol·(g·h)^-1和62.4 μmol·(g·h)^-1,比原始菌株Rhodocossus erythropolis DS-3的35.2 μmol·(g·h)^-1、 21.3 μmol·(g·h)^-1和25.5 μmol·(g·h)^-1提高了1.5倍左右.其中Gordonia sp. CRA和Gordonia sp. CRB在以DBT为唯一硫源的培养基中几乎不生长,无法降解DBT;而Gordonia sp. CRC同表达完整DBT脱硫酶的Gordonia sp. CRABC一样,在以DBT为唯一硫源的培养基中生长良好,亦能降解大部分DBT（84％）.这表明催化BT和DBT前两步脱硫反应的BT单加氧酶和DBT单加氧酶是负责底物识别的关键酶,催化BT和DBT后两步脱硫反应的酶功能相似,通过比较这2种单加氧酶的氨基酸序列差异,即可预测其作用的活性位点.     

功能菌接种对石油污染土壤修复效果及微生物群落的影响

利用定向筛选驯化的二苯并噻吩（DBT）降解菌对石油污染土壤进行为期40 d的土壤模拟培养试验,研究了功能菌（红球菌,Rhodococcus sp. ZYL-1）接种对石油污染土壤中DBT的降解效果,结合16S rDNA高通量测序及生物信息学分析,解析了功能菌接种对土壤细菌群落演替的影响。结果表明：在25 ℃的暗箱培养过程中,红球菌添加显著提高了DBT污染土壤的降解率（P<0.001）,生物接种处理组（BIOEIF）的DBT降解主要发生在培养前10 d,培养结束后DBT降解率接近60%,比依赖土壤土著微生物的自然衰减组（NAT）降解率提升10%以上;对比BIOEIF组和NAT组土壤培养过程中细菌群落组成,BIOEIF组香农多样性指数和系统发育指数显著低于NAT组,但接种红球菌未对DBT污染土壤的细菌群落组成造成显著影响,Micromonospora、Bacillus、unclassified_f_Planococcaceae为污染土壤培养过程中的优势菌属,而接种的红球菌并未成为优势菌属;网络分析表明,功能菌接种显著提升污染土壤微生物的DBT关键降解菌属类群,通过强化土壤土著微生物Shimazuella协同降解DBT,进而提升了石油烃污染土壤的修复效果,Shimazuella可能是参与DBT的代谢的关键微生物。所接种的功能菌（红球菌）可协同提高土著微生物对于石油污染土壤DBT的生物降解,具有较高的土壤修复应用潜力。

     

苯并噻吩脱硫菌株的筛选及脱硫活性研究

从孤岛油田油浸土样中筛选到1株能降解苯并噻吩(BT)的脱硫菌,经初步鉴定该菌为戈登氏菌属(Gordona sp.).实验证明:该菌能以类似于4S途径脱除BT及其衍生物中的硫,但是不能脱除二苯并噻吩(DBT)及其衍生物中的硫.GC-MS分析表明该途径的终产物为邻羟基苯乙醛或其异构体苯并呋喃.在以BT为唯一硫源的培养基中30℃培养48h,Gordona sp.C-6能降解0.15mmol/L的BT,终产物占发酵培养基中BT加入量的50%,其余BT在有氧培养过程中挥发.通过Matlab拟合曲线确定以邻羟基苯乙酸为标准品进行产物定量检测的方法.     

二苯并噻吩降解菌筛选及16S rDNA序列分析

以DBT(dibenzothiophene,二苯并噻吩)为模型化合物,分离得到了一株能降解DBT的微生物HB2,应用HPLC对其脱硫特性进行了检测。应用PCR技术克隆到16SrDNA片段,核苷酸序列分析结果表明,该菌的16SrDNA全序列与假单胞菌存在98%的同源性,初步确定该菌在微生物系统发育学上的地位。     

利用休止细胞法选择性脱除燃料油中有机硫

红球菌(Rhodococcus sp .)FS-1菌株能通过专一性断裂C—S键的途径脱除二苯并噻吩 (DBT)中的有机硫作为自身生长需要的硫源 ,从而降低油品中的硫含量 .本文利用该菌的休止细胞对DBT和柴油的脱硫活力进行了研究 .结果表明 ,该菌株对DBT及柴油中的有机硫均有良好的选择性脱除作用 .DBT浓度为0.5～1.0mmol/L、油水比例为 1:5时 ,脱硫效果最佳 .采用二次脱硫法可使柴油脱硫率达85%以上 ,证明FS-1能有效脱除柴油中的有机硫 .烃质组分的气相色谱分析表明 ,FS-1作用前后的柴油烃类组分基本没有改变 ,说明该菌的脱硫作用是特异性针对硫原子的 ,不会破坏柴油的有效成分 .     

聚乙烯醇包埋石油脱硫菌UP-2的研究

以筛选出的具有脱硫能力的施氏假单胞菌UP-2为固定化研究对象,二苯并噻吩(DBT)为生物催化脱硫模型化合物,考察了脱硫菌UP-2的固定化操作条件和固定化细胞使用条件.结果表明,当包埋剂聚乙烯醇(PVA)浓度为10%、添加剂海藻酸钠(SA)浓度为0.2%、液菌比为201时,在4℃、含有1%CaCl2的饱和硼酸中交联24h后,可以得到脱硫性能很好的固定化细胞小球;在30℃、pH值为7.0的体系中反应6d,可将浓度为576mg/L的DBT降解70%左右,固定化细胞降解DBT的比活性由未固定化细胞的0.49mmol/gdw增加到6.39mmol/gdw,使用寿命高达800h以上.     

基于Cx-DBT的微生物脱硫研究进展

烷基取代二苯并噻吩Cx-DBT作为微生物脱除煤中有机硫研究的模型化合物在理论和研究实践中可以获得足够的支持，而传统的脱硫研究仅仅是围绕单一模型化合物DBT展开的。对已经用于研究工作的模型化合物和基于Cx-DBT的微生物脱硫研究进展进行了论述，分析了取代基、分子形态和温度等参数对于微生物脱除Cx—DBT结构中有机硫活性的影响，介绍了不对称取代Cx-DBT脱硫时的第一个C—S键断裂的优先模式和微生物脱硫酶系统的不同识别特征。     

施氏假单胞菌对二苯并噻吩的降解

从胜利油田油井附近土壤中筛选到 1 株施氏假单胞菌(Pseudomonas stutzeri UP1),可把二苯并噻吩(DBT)降解成水溶性硫化物,静息细胞实验证实该菌株细胞内存在能够降解DBT的酶系,降解过程的某些中间产物及终产物与已知的Kodama 路线相同,表明此菌株对DBT的降解是以 Kodama 路线进行的.     

Ni/Ce-Y沸石的制备及其对二苯并噻吩吸附研究

研究了负载二元金属NiCe-Y沸石的制备以及吸附二苯并噻吩的动力学性能。试验考察了二元金属Ni、Ce离子交换顺序、负载溶液方面对吸附剂制备的影响,初步得到了制备负载二元金属NiCe-Y吸附剂的优化条件。对NiCe-Y吸附剂进行脱硫性能评价及动力学分析,结果表明,负载二元金属NiCe-Y吸附剂比负载单一金属的Ni-Y吸附量提高31%,比Ce-Y吸附剂提高16%。     

Effects of nicotinamide and riboflavin on the biodesulfurization activity of dibenzothiophene by Rhodococcus erythropolis USTB-03

Rhodococcus erythropolis USTB-03 is a promising bacterial strain for the biodesulfurization of dibenzothiophene (DBT) via a sulfurspecific pathway in which DBT is converted to 2-hydroxybiphenyl (2HBP) as an end product. The effects of nicotinamide and riboflavin on the sulfur specific activity (SA) of DBT biodesulfurization by R. erythropolis USTB-03 were investigated. Both nicotinamide and riboflavin were found to enhance the expression of SA, which was not previously reported. When R. erythropolis USTB-03 was grown on a medium containing nicotinamide of 10. 0 mmol or riboflavin of 50. 0 μnol, SA was raised from 68. 0 or so to more than 130 mmol 2HBP/(kg dry cells. h). When R. erythropolis USTB-03 was grown in the presence of both nicotinamide of 5. 0 mmol and riboflavin of 25. 0 μmol, SA was further increased to 159. 0 mmol 2HBP/(kg dry cells. h). It is suggested that the biological synthesis of reduced form of flavin mononucleotide (FMNH2), an essential coenzyme for the activities of biodesulfurization enzyme Dsz C and A, might be enhanced by nicotinamide and riboflavin, which was responsible for the increased SA of R. erythropolis USTB-03.     

Desulfurization of dibenzothiophene by a newly isolated Corynebacterium sp. ZD-1 in aqueous phase

IntroductionEmissionofsulfur oxidestotheatmospherethroughcombustionoffossilfuelisamainreasonthatcausesseriousenvironmentalproblemsuchasacidrain.Asthecrisisofenergysourcebecomeworse ,mineoffossilfuelwithhighsulfurcontentsisinevitable.Butthedemandofmiddle d…     

Desulfurization of dibenzothiophene（DBT） by a novel strain Lysinibacillus sphaericus DMT-7 isolated from diesel contaminated soil

A new bacterial strain DMT-7 capable of selectively desulfurizing dibenzothiophene (DBT) was isolated from diesel contaminated soil. The DMT-7 was characterized and identified as Lysinibacillus sphaericus DMT-7 (NCBI GenBank Accession No. GQ496620) using 16S rDNA gene sequence analysis. The desulfurized product of DBT, 2-hydroxybiphenyl (2HBP), was identified and confirmed by high performance liquid chromatography analysis and gas chromatography-mass spectroscopy analysis respectively. The desulfurization kinetics revealed that DMT-7 started desulfurization of DBT into 2HBP after the lag phase of 24 hr, exponentially increasing the accumulation of 2HBP up to 15 days leading to approximately 60% desulfurization of the DBT. However, further growth resulted into DBT degradation. The induced culture of DMT-7 showed shorter lag phase of 6 hr and early onset of stationary phase within 10 days for desulfurization as compared to that of non-induced culture clearly indicating the inducibility of the desulfurization pathway of DMT-7. In addition, Lysinibacillus sphaericus DMT-7 also possess the ability to utilize broad range of substrates as sole source of sulfur such as benzothiophene, 3,4-benzo DBT, 4,6-dimethyl DBT, and 4,6-dibutyl DBT. Therefore, Lysinibacillus sphaericus DMT-7 could serve as model system for efficient biodesulfurization of diesel and petrol.     

固相微萃取-液相色谱法测定水中二苯并噻吩

分别采用高效液相色谱与固相微萃取-高效液相色谱法建立了水中微量二苯并噻吩的分析方法,结果表明固相微萃取对二苯并噻吩具有非常明显的富集作用。实验得到固相微萃取技术最佳检测条件为:吸附时间20min,解析时间3min,色谱流动相甲醇/水=90/10,方法的变异系数为2.85%,加标回收率102.4%,检测限0.03μg/mL。     

Contribution of the Kodama and 4S pathways to the dibenzothiophene biodegradation in different coastal wetlands under different C/N ratios

Dibenzothiophene (DBT) degradation mechanisms and the transformation of pathways during the incubation of three types of coastal sediments with C/N ratios ranging from 1 to 9 were investigated. The DBT degradation efficiencies were clearly improved with increasing C/N ratio in reed wetland sediments, tidal wetlands sediments and estuary wetland sediments. The quantitative response relationships between DBT degradation rates and related functional genes demonstrate that the Kodama pathway-related gene groups were dominant factors at low C/N ratios, while the 4S-related gene groups mainly determined the degradation rate when the C/N ratio was up to 5. Network analysis also shows that the pathway shifts from the Kodama pathway to the 4S pathway occurred through changes in the connections between functional genomes and rates. Furthermore, there were competition and collaboration between the Kodama and 4S pathways. The 4S pathway-related bacteria were more active in estuary wetland sediments compared with reed wetland sediments and tidal wetland sediments. The higher degradation efficiency in estuary wetland sediments may indicate the greater participation of the 4S pathway in the DBT biodegradation reaction. And the effects of ring cleavage of Kodama pathway caused more complete metabolizing of DBT.     

海洋耐镉微生物筛选及其生物学特性

从重金属污染的近海域筛选到一株对镉具有较强抗性和富集能力的微生物Cd20002,该菌株在液体培养基中的最大耐受浓度为1 000 mg/L,在镉浓度达到200 mg/L时,其吸附率高达93.4%,根据形态特征和生理生化鉴定及16S rRNA测序鉴定为蓝杆菌。通过在不同的镉浓度环境对蓝杆菌Cd20002进行培养,测定其生长曲线,并优化菌体生长的条件及其吸附能力。结果表明,蓝杆菌Cd20002既能在重金属镉环境中生存,且富镉能力较强,对水产环境中重金属的生物清排具有重要意义。