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 .     

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.     

二苯并噻吩脱硫微生物菌种的筛选与活性

从被石油污染的土壤中筛选出1株能对二苯并噻吩(DBT)进行高效脱硫的微生物菌种,初步鉴定为红串红球菌USTB-03(RhodococcuserythropolisUSTB-03).该菌种可以按特异性脱硫途径(简称4S途径)将DBT转化为2-羟基联苯(2HBP)和亚硫酸作为最终脱硫产物.在葡萄糖、甘油和乙醇分别作为微生物生长的唯一碳源时,葡萄糖是支持该菌生长和提高其脱硫比活性的较好碳源,使培养出的微生物对DBT的脱硫比活性达到了68.63mmol2HBP/(kg·h).该菌株还可以对4,6-二甲基二苯并噻吩(4,6-DMDBT)进行脱硫.     

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.     

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.     

沼泽红假单胞菌H3对酸性红B2GL染料的厌氧脱色和降解作用

从印染厂污泥中分离到一株沼泽红假单胞菌（ＲｈｏｄｏｐｓｅｕｄｏｍｏｎａｓｐａｌｕｓｔｒｉｓＨ３），在光照厌氧条件下该菌生长细胞可将１００ｍｇ／Ｌ酸性红Ｂ２ＧＬ染料去除到３０ｍｇ／Ｌ．完整细胞脱色的最适条件为ｐＨ７０，温度３０℃，细胞浓度２０—２５ｍｇ／ｍＬ（湿重）．低浓度的阳离子对脱色影响不大．在通Ａｒ气使严格厌氧和加有还原性辅酶Ｉ的条件下无细胞提取液的脱色活性最高，比活率为１５４×１０－２ｍｇ／（ｍｇ·ｈ）．根据降解产物的分析，推断了该菌对酸性红染料的降解代谢途径．     

苯酚降解菌的分离及降解特性研究

从扬子乙烯集团废水处理系统曝气池中的活性污泥驯化分离得到一株能快速降解苯酚的菌株,初步鉴定其为假单胞菌属菌株。该菌株在5℃-35℃范围内时都可以有效降解并矿化200mg／L的苯酚,最适宜的生长温度为25℃左右;菌株在pH为5～9范围内可以降解200mg／L的苯酚,偏碱性的条件下比酸性条件更适合细菌生长;培养过程中振荡速率大于120r／m时降解速率最大。当苯酚的初始浓度超过1000mg／L时,降解菌的生长受到抑制,不能有效降解苯酚。     

氯嘧磺隆降解菌的筛选及对污染土壤的生物修复

采用富集培养方法从江苏某激素研究所污水处理池排污口污泥中分离得到1株氯嘧磺隆降解菌,经菌株形态学特征和26S rDNA序列分析,鉴定为胶红酵母菌（Rhodotorula mucilaginosa ）.经降解条件优化,菌株对含100mg/L氯嘧磺隆的无机盐培养基中氯嘧磺隆的最佳降解条件为：接种量2.5%,培养温度28℃,pH 6.0,培养5d后降解率为87.33%;在氯嘧磺隆初始浓度为10mg/kg（干土）的模拟污染土壤中,菌株最佳降解条件为：接种量2.5%,温度25℃,pH 6.0,土壤含水量30%,静息培养30d后降解率为90.74%.土壤修复实验结果表明,施加胶红酵母菌后减轻了氯嘧磺隆对小麦幼苗的药害,在氯嘧磺隆浓度为10mg/kg的土壤中投加降解菌后,小麦的出苗率、株高、根长及鲜重均明显高于未投加降解菌的对照组（P < 0.05）.     

外加碳源对红球菌IcdP1降解荧蒽的特性研究

从受多环芳烃(PAHs)污染的河道底泥中筛选分离、纯化得到一株荧蒽降解菌,经形貌分析和16S r DNA序列分析后确定该菌株为红球菌属Rhodococcus sp.,命名为Icd P1。之后对该菌株降解荧蒽性能进行了系统研究。结果表明,当荧蒽浓度为50 mg/L时,Icd P1在培养42 d后对荧蒽降解率可到达31.69%。投加"蒽-菲"混合物、葡萄糖、麦芽糖、腐殖酸(HA)等外加碳源可以不同程度强化Icd P1降解荧蒽的能力,其中投加葡萄糖对红球菌降解荧蒽的强化效果最好,且荧蒽去除率和红球菌生物量之间的线性相关关系最好,说明葡萄糖最适合作为该红球菌降解荧蒽的共代谢基质。动力学分析表明,无论是否添加葡萄糖作为外加碳源,红球菌Icd P1对荧蒽的降解都符合一级动力学降解模型,但葡萄糖的加入大大提高了Icd P1菌对荧蒽的降解速率。     

红球菌J-5菌株降解聚乙烯醇的研究

从土壤样品中分离到一株能以聚乙烯醇(PVA)为唯一碳源、能源生长的纯菌株,经生理生化鉴定和16S rRNA基因序列分析,该菌株属于红球菌属(Rhodococcus sp.),对该菌株降解PVA的特性进行了研究.结果表明,该菌株降解PVA的适宜条件为10000mg/L PVA,25℃,摇床培养;此外,添加0.1%的酵母膏可以明显促进PVA的降解.在该条件下通气培养7d,可以降解58%的PVA,通过红外光谱分析PVA的代谢产物,说明PVA降解的中间产物带有羰基.     

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（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.     

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.     

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

IntroductionEmissionofsulfur oxidestotheatmospherethroughcombustionoffossilfuelisamainreasonthatcausesseriousenvironmentalproblemsuchasacidrain.Asthecrisisofenergysourcebecomeworse ,mineoffossilfuelwithhighsulfurcontentsisinevitable.Butthedemandofmiddle d…     

硫酸盐还原菌的分离和生理特性研究

从受氯碱化工废水严重污染的湖北鸭儿湖1号氧化塘底泥中分离获得了硫酸盐还原菌,研究了其生理特性以及温度、pH、盐度、Fe2+和硫化物等环境因子对其的影响。结果显示,该硫酸盐还原菌营厌氧生活,革兰氏染色呈阴性,可以乳酸钠为碳源(电子供体),硫酸盐等为硫源(电子受体),该过程中有H2S的产生。该菌在35℃、pH7.0、0.7%的盐度、0.5g/LFe2+和不含硫化物等条件下,达到最佳生长状态。     

二苯噻吩的微生物脱硫及其在脱煤中有机硫的应用

评述了煤中有机硫的模型化合物二苯噻吩的微生物脱硫最新进展，对二苯噻吩的特异性脱硫的微生物和机理作了较新的介绍并提出了独到的看法，指出了当今存在的问题和今后的研究方向及其在脱煤中有机硫的应用前景。     

1株异养反硝化硫细菌的分离鉴定及代谢特性

工业废水中的有机物、硫化物和含氮化合物是废水处理面临的重大挑战.利用生物技术,可以实现在废水处理系统中同时去除这3种污染物.在污水处理反应器中,分离出1株异养反硝化硫细菌HDD1.基于16S rRNA基因的系统进化分析和生理特征显示菌株HDD1为Thauera属的一个种.菌株HDD1能够利用乙酸盐和硫化物作为电子供体,硝酸盐作为电子受体进行呼吸作用.在15h之内,CH3COO-(300 mg·L~(-1))、S2-(200 mg·L~(-1))和NO-3(487 mg·L~(-1))被完全代谢去除.扫描电子显微镜和能量色散谱结果显示硫化物氧化的主要产物为单质硫.菌株HDD1能够同时应用于工业废水处理和硫元素的资源化回收.     

Experimental and modeling investigations on the unexpected hydrogen sulfide rebound in a sewer receiving nitrate addition: Mechanism and solution

Biogenic hydrogen sulfide is an odorous, toxic and corrosive gas released from sewage in sewers. To control sulfide generation and emission, nitrate is extensively applied in sewer systems for decades. However, the unexpected sulfide rebound after nitrate addition is being questioned in recent studies. Possible reasons for the sulfide rebounds have been studied, but the mechanism is still unclear, so the countermeasure is not yet proposed. In this study, a lab-scale sewer system was developed for investigating the unexpected sulfide rebounds via the traditional strategy of nitrate addition during 195-days of operation. It was observed that the sulfide pollution was even severe in a sewer receiving nitrate addition. The mechanism for the sulfide rebound can be differentiated into short-term and long-term effects based on the dominant contribution. The accumulation of intermediate elemental sulfur in biofilm resulted in a rapid sulfide rebound via the high-rate sulfur reduction after the depletion of nitrate in a short period. The presence of nitrate in sewer promoted the microorganism proliferation in biofilm, increased the biofilm thickness, re-shaped the microbial community and enhanced biological denitrification and sulfur production, which further weakened the effect of nitrate on sulfide control during the long-term operation. An optimized biofilm-initiated sewer process model demonstrated that neither the intermittent nitrate addition nor the continuous nitrate addition was a sustainable strategy for the sulfide control. To minimize the negative impact from sulfide rebounds, a (bi)monthly routine maintenance (e.g., hydraulic flushing with nitrate spike) to remove the proliferative microorganism in biofilm is necessary.