2,4-二氯酚降解菌的分离及其特性

从３个不同地点采集的混合土壤中，分离到２株可利用２，４二氯酚为唯一碳源和能源的假单胞菌（Ｐｓｅｕｄｏｍｏｎａｓｓｐ．）ＤＣＰ１和ＤＣＰ２．在此基础上，研究探索了菌株ＤＣＰ１降解２，４二氯酚的影响因素和降解过程以及其遗传特性．结果表明：菌株ＤＣＰ１可降解２，４二氯酚，并有氯离子、邻氯酚和醌积累；经检测该菌株中有一条迁移速率比ｐＵＣ１８小的质粒．     

Isolation and characterization of a profenofos degrading bacterium

Profenofos, a well known organophosphate pesticide, has been in agricultural use over the last two decades for controlling Lepidopteron pests of cotton and tobacco crops. In this study, a bacterial strain, OW, was isolated from a long term profenofos exposed soil by an enrichment technique, and its ability to degrade profenofos was determined using gas chromatography. The isolated strain OW was identified as Pseudomonas aeruginosa according to its physiological and biochemical properties, and the analysis of its 16S rRNA gene sequence. The strain grew well at pH 5.5–7.2 with a broad temperature profile. Bioremediation of profenofos-contaminated soil was examined using soil treated with 200 g/g profenofos, which resulted in a higher degradation rate than control soils without inoculation. In a mineral salt medium (FTW), removal in the level of profenofos of 86.81% was obtained within 48 h of incubation. The intermediates of profenofos metabolism indicated that the degradation occurred through a hydrolysis mechanism, and one of the metabolites was found to be 4 bromo-2-cholorophenol (BCP) which in turn was also mineralized by the strain. The results of this study highlighted the potentiality of P. aeruginosa as a biodegrader which could be used for the bioremediation of profenofos contaminated soil.     

Isolation and characterization of heterotrophic nitrifying bacteria in MBR

The study presented the method for isolating the heterotrophic nitrifiers and the characterization of heterotrophic nitrification. Continuous tests via a membrane bioreactor (MBR) were operated under the controlled conditions to proliferate the nitrifiers. Heterotrophic nitrifying bacteria were isolated from the system in which the efficiency of total nitrogen(TN) removal was up to 80%. Since no autotrophic ammonium and nitrite oxidizers could be detected by fluorescence in situ hybridization(FISH), oxidized-N production was unlikely to be catalyzed by autotrophic nitrifiers during the heterotrophic nitrifiers‘ isolation in this study. The batch test results indicate that the isolated heterotrophic bacteria were able to nitrify. After 3 weeks incubation, the efficiencies of the COD removal by the three isolated bacterial strains B1,B2 and B3 were 52.6%, 71.7%, and 77.7%, respectively. The efficiencies of the TN removal by B1, B2, and B3 were 35.6%, 61.2% and 68.7%, respectively.     

诺氟沙星降解菌NOR-36的分离筛选及降解特性研究

利用微生物分离筛选技术从医药厂废水池活性污泥中分离得到一株可以以哌嗪为一级基质共代谢降解诺氟沙星的菌株.经生理生化鉴定和16S rDNA基因序列同源性分析,将此菌株初步鉴定为山羊葡萄球菌(Staphylococcus caprae),命名为NOR-36.当菌株NOR-36在温度为30℃、pH 7.0、诺氟沙星起始浓度为5 mg·L~(-1),哌嗪与诺氟沙星浓度比为5∶1时培养10 d降解率可达92.6%.而在相同的实验条件下,不加一级基质的空白实验中诺氟沙星的降解率最高仅为8.5%.可知菌株NOR-36以哌嗪为一级基质对诺氟沙星进行共代谢生物降解,其对诺氟沙星的降解率显著提高.采用高效液相色谱-质谱联用仪分析确定菌株降解诺氟沙星的过程中产生3种主要的中间代谢产物,分子式分别为:C_(16)H_(18)FN_3O_4(产物1)、C_(14)H_(16)FN_3O_3(产物2)、C_(12)H_(11)FN_2O_3(产物3).分析中间产物结构可知:NOR-36首先在共代谢关键酶作用下使诺氟沙星哌嗪环上碳氮键断裂,并且中间位置碳原子被氧原子氧化,加氧形成羰基,得到产物1;之后产物1继续被氧化,碳氮键继续断裂,—COCH_3基团被去除生成产物2,最后产物2经氧化后,—CH_2CH_2NH_2基团被去除,最终得到产物3.     

Isolation and preliminary characterization of a 3-chlorobenzoate degrading bacteria

A study was conducted to compare the diversity of 2-, 3-, and 4-chlorobenzoate degraders in two pristine soils and one contaminated sewage sludge. These samples contained strikingly different populations of mono-chlorobenzoate degraders. Although fewer cultures were isolated in the uncontaminated soils than contaminated one, the ability of microbial populations to mineralize chlorobenzoate was widespread. The 3- and 4-chlorobenzoate degraders were more diverse than the 2-chlorobenzoate degraders. One of the strains isolated from the sewage sludge was obtained. Based on its phenotype, chemotaxonomic properties and 16S rRNA gene, the organism S-7 was classified as Rhodococcus erythropolis. The strain can grow at temperature from 4 to 37℃. It can utilize several （halo）aromatic compounds. Moreover, strain S-7 can grow and use 3-chlorobenzoate as sole carbon source in a temperatures range of 10-30℃ with stoichiometric release of chloride ions. The psychrotolerant ability was significant for bioremediation in low temperature regions. Catechol and chlorocatechol 1,2-dioxygenase activities were present in cell free extracts of the strain, but no （chloro）catechol 2,3- dioxygenase activities was detected. Spectral conversion assays with extracts from R. erythropolis S-7 showed accumulation of a compound with a similar UV spectrum as chloro-cis,cis-muconate from 3-chlorobenzoate. On the basis of these results, we proposed that S-7 degraded 3-chlorobenzoate through the modified ortho-cleave pathway.     

Isolation and characterization of deodorizing bacteria for organic sulfide malodor

Strain JII screened out from different odor origins can efficiently degrade methyl mercaptan and ethanethiol whereas has no ability to remove dimethyl sulfide. The results indicated that the strain JII breaks only the C-SH bond. The optimum temperature and pH of JII are 20-30℃ and 6.0-8.3 respectively. A systematic identificatio nmethod-16S rDNA gene sequence comparison, for deodorizing bacteria was carried out. The 16S rDNA gene sequence analysis of strain JII showed the highest level of 97 % homology to Rape rhizosphere.     

一株降解微囊藻毒素的缺陷短波单胞菌的分离鉴定及降解特性研究

从太湖水华腐烂蓝藻中筛选出一株能够降解MC-LR的细菌,将其编号为Q3.经形态、生理生化及16S rRNA基因序列分析鉴定为缺陷短波单胞菌(Brevundimonas diminuta).研究发现,在实验条件下该菌能以MC-LR为唯一碳源和氮源生长;7 d内可将初始浓度为0.96 mg·L-1的MC-LR降解为0.37 mg·L-1,降解效率达到61.5%.同时,本研究首次发现缺陷短波单胞菌能够降解藻毒素,并且在此菌种中扩增出了降解过程的关键基因mlr A,推测该菌可能与已报道的Sphingomonas sp.ACM-3962等菌具有相同的降解机制.     

苯酚降解菌CM-HZX1菌株的分离、鉴定及降解性能研究

从污水处理厂的活性污泥中分离出一株以苯酚为唯一碳源生长的高效降解苯酚菌CM-HZX1.通过形态特征、生理生化及16S r DNA基因序列分析,初步鉴定菌株属于红球菌(Rhodococcus sp.),16S r DNA在Gene Bank的登录号为KM014567.实验结果表明,菌株CM-HZX1培养及降解苯酚的最适条件为p H=7.0,温度30℃,转速为150 r·min~(-1).该菌株能耐受4%的盐度,适应性强.0.5 g·L~(-1)苯酚在24 h时的降解率可达93.6%,1.5 g·L~(-1)苯酚在48 h时的降解率在90%以上.研究表明,该菌株在处理工业含酚废水方面具有广阔的应用前景.     

脂肽类生物表面活性剂产生菌的分离及特性研究

从石油污染土壤中分离筛选获得一株产生生物表面活性剂菌株Y8A,经生理生化实验、16S rDNA序列分析等将其鉴定为芽孢杆菌属（Bacillus sp.）.Y8A能在22h内将发酵液的表面张力从68.3mN·m-1降到23.5 mN·m-1.经TLC和傅立叶红外光谱分析, 菌株Y8A产生的生物表面活性剂为脂肽类.20mg·L-1 Ca2+和Fe2+能显著促进其生长和表面活性剂的产生;菌株Y8A在20～30℃,pH 5～12范围内产生表面活性剂的能力较强;LB培养基中添加1%乳糖对生长的影响不大,但能够明显促进Y8A产生生物表面活性剂,而葡萄糖、蔗糖抑制表面活性剂的产生.Y8A能够促进石油降解菌Y1D和F11对石油的降解和功夫菊酯降解菌ZZH对功夫菊酯的生物降解.     

海洋产毒甲藻——塔玛亚历山大藻(Alexandrium tarmarense)基因组DNA微卫星位点分离

采用FIASCO(Fast Isolation by AFLP of Sequences Containing repeats site)方法富集、分离海洋产毒甲藻——塔玛亚历山大藻基因组DNA微卫星位点。结果发现,测序分析的22个克隆中全部含有微卫星位点,插入片段大小为175-687 bp,微卫星DNA的重复数最低4次,最高27次,平均11次;在全部的39个微卫星位点中,其中25个属于完美重复型,13个不完美重复型,1个复合型。为塔玛亚历山大藻系统进化生物学、分子生态学研究提供了很好的标记位点。     

2株高效溶赤潮异湾藻放线菌的鉴定及溶藻特性

从武汉周边土壤中分离得到2株具有强溶藻能力的放线菌,命名为B-10和G-11.研究了2株菌对赤潮异弯藻(Heterosigma akashiwo)、锥状斯克里普藻(Scrippsiella trochoidea)、微小原甲藻(Prorocentrum minimum)、金藻(Golden algae)的溶藻效果及溶藻方式,通过形态学特征、培养特征、生理生化鉴定、16S rDNA序列分析对其进行鉴定.研究结果表明,株放线菌对赤潮异湾藻和锥状斯克里普藻都有较好的溶藻效果,对微小原甲藻和金藻的生长也有抑制作用,且在传代5次后能够保持稳定的溶藻效果, 按1％的体积比接种5d后B-10和G-11对异湾藻的去除率分别为90%和99.44%.通过溶藻方式研究,发现2菌株都是通过释放胞外代谢物溶藻,且胞外代谢产物对温度敏感,当温度达到100℃时会使2种溶藻物质失活.经形态学特征、生理生化鉴定及16S rDNA序列分析结果,可初步判断,B-10菌株属于链霉菌属的细黄链霉菌(Streptomyces microflavus)类群,菌株G-11属于链霉菌属的变异链霉菌(Streptomyces variabilis )类群.     

磺胺二甲基嘧啶(SM2)高效降解菌J2的分离筛选及降解特性研究

从城市污水处理厂活性污泥中分离得到一株能以磺胺二甲基嘧啶(SM2)为唯一碳源的菌株,经生理生化鉴定和16S rRNA基因序列同源性分析,将此菌鉴定为蜡样芽孢杆菌(Bacillus cereus),命名为J2.不同条件下的降解特性研究结果表明,J2菌株具有极高的SM2耐受性(100 mg·L~(-1))、较广的温度(20～30℃)和pH(6～8)适用范围;在温度30℃、pH=8.0、初始OD_(600)=0.1、SM2起始浓度为50 mg·L~(-1)的条件下,J2菌株在36 h内对SM2的降解率可达100%,降解效率远超目前已报道的其他SM2降解菌株,展现出了良好的应用潜力.J2菌株降解SM2过程中产生了5种主要中间代谢产物,分析推断其降解SM2的途径分为两条:①磺胺二甲基嘧啶分子首先在酶促反应作用下脱除SO_2,生成嘧啶环和苯胺环自由基,这两种自由基再经过环间耦合生成N-(4,6-二甲基嘧啶-2基)-1,4-二苯胺,该分子中的C—N键在活性氧物种的作用下断开生成苯胺和2-氨基-4,6-二甲基嘧啶;②在漆酶的作用下N~4键断裂,产生N-(3,5-二甲基嘧啶)-苯磺酰胺,之后N-(3,5-二甲基嘧啶)-苯磺酰胺的N—S键断裂,进一步形成2-氨基-4,6-二甲基嘧啶和苯亚砜.     

Isolation and characterization of gasoline-degrading bacteria from gas station leaking-contaminated soils

The effects of culture conditions in vitro and biosurfactant detection were studied on bacterial strains capable of degrading gasoline from contaminated soils near gas station. The main results were summarized as follows. Three bacteria （strains Q10, Q14 and Q18） that were considered as efficiently degrading strains were isolated and identified as Pseudomonas sp., Flavobaeterium sp. and Rhodococcus sp., respectively. The optimal growth conditions of three bacteria including pH, temperature and the concentration of gasoline were similar. The reduction in surface tension was observed with all the three bacteria, indicating the production of biosurfactant compounds. The value of surface tension reduced by the three strains Q10, Q14 and Q18 was 32.6 mN.m, 12.4 mN. m and 21.9 mN.m, respectively. Strain Q10 could be considered as a potential biosurfactant producer. Gasoline, diesel oil, benzene, toluene, ethylbenzene and xylene （BTEX） could easily be degraded by the three isolates. The consortium was more effective than the individual cultures in degrading added gasoline, diesel oil, and BTEX. These results indicate that these strains have great potential for in situ remediation of soils contaminated by gas station leaking.     

降解甲基对硫磷的节杆菌(Arthrobacter sp.)L4菌株的分离和降解特性研究

从生产甲基对硫磷的山东华阳农药厂污水曝气池中,分离到一株能以甲基对硫磷及其降解中间产物对硝基苯酚为唯一碳源生长的细菌L4菌株.经16S rRNA基因序列分析,该菌株被鉴定为节杆菌属(Arthrobacter sp.).用气相色谱法和分光光度法分析L4菌株的降解性能,结果表明,L4在5h内对50mg·L-1的甲基对硫磷和对硝基苯酚的降解率分别为85%和98%,对其它有机磷农药也有良好的降解效果.L4的最适培养条件为pH值7、30℃、接种量30%.     

Isolation and characterization of facultative mixotrophic ammonia-oxidizing bacteria from constructed wetlands

Autotrophic ammonia-oxidizing bacteria (AOB) have been widely studied in constructed wetlands systems, while mixotrophic AOB have been less thoroughly examined. Heterotrophic bacteria were isolated from wastewater and rhizospheres of macrophytes of constructed wetlands, and then cultivated in a mixotrophic medium containing ammonium and acetic acid. A molecular characterization was accomplished using ITS-PCR amplification, and phylogenetic analysis based on 16S rRNA gene sequences. Results showed the presence of 35 bacteria, among 400 initially heterotrophic isolates, that were able to remove ammonia. These 35 isolates were classified into 10 genetically different groups based on ITS pattern. Then, a collection of 10 isolates were selected because of their relatively high ammonia removal efficiencies (ARE ≥ 80%) and their phylogenetic diversity. In conditions of mixotrophy, these strains were shown to be able to grow (increase of optical density OD₆₆₀ during incubation with assimilation of nitrogen into cellular biomass) and to oxidize ammonia (important ammonia oxidation efficiencies, AOE between 79% and 87%). Among these facultative mixotrophic AOB, four isolates were genetically related to Firmicutes (Bacillus and Exiguobacterium), three isolates were affiliated to Actinobacteria (Arthrobacter) and three other isolates were associated with Proteobacteria (Pseudomonas, Ochrobactrum and Bordetella).     

一株多氯联苯降解菌的筛选鉴定及降解性能研究

从长期受有机污染的土壤中驯化、分离出1株高效多氯联苯降解菌,经生理生化和16S r DNA测序鉴定为铜绿假单胞菌(Pseudomonas aeruginosa),用其对PCB77进行降解性能研究.结果表明,该菌株能够以PCB77为唯一碳源生长,在培养温度为30℃、p H值7.5、PCB77浓度1.0 mg·L-1、接菌量2 m L(OD600=1.0)、摇床转速150 r·min-1的培养条件下,7 d后PCB77的降解率为49.6%;菌株在外加相同浓度联苯和邻苯二甲酸时,降解效率分别提高到58.5%和53.8%,而加入苯甲酸时,降解率为40.8%;外加重金属Cr6+和Pb2+对菌株降解PCB77均有显著的抑制作用;当重金属浓度较低时抑制作用不显著,而浓度较高时有明显的抑制作用,且Cr6+对菌株降解能力的抑制强于Pb2+.     

噻嗪酮降解菌BF3的分离、鉴定及降解特性

分离到1株能降解噻嗪酮的细菌BF3,通过生理生化试验和16S rRNA基因同源性序列分析,初步鉴定其属于副球菌属(Paracoccus sp.).该菌在接种量为2%的条件下,7d内对50mg/L噻嗪酮的降解率为68.2%.其最适降解pH值为7.0,降解最适温度为30℃.BF3对低浓度(1,10,30,50mg/L)的噻嗪酮有较好的降解效果,但对高浓度(100,200mg/L)的噻嗪酮降解效果一般,降解速率与起始接种量呈正相关.需氧量试验表明,BF3在通气良好的状况下降解速率较高.     

苯胺降解菌的分离和特性研究

从活性污泥中分离到一株细菌ＡＮ３，能以苯胺为唯一碳源、氮源和能源生长．经鉴定为食酸丛毛胞菌（Ｃｏｍａｍｏｎａｓａｃｉｄｏｖｏｒａｎｓ）．该菌株可以在高达５０００ｍｇ／Ｌ以上的苯胺中生长．当苯胺浓度为２０００ｍｇ／Ｌ左右时，经３天培养即可全部被降解．该菌株还可以利用乙酰苯胺，但不能利用其他取代类苯胺化合物．ＡＮ３菌的生长和降解苯胺的最适温度为３０℃和ｐＨ为７０．９种金属离子对该菌的生长和苯胺的降解均有不同程度抑制作用，尤以Ａｇ＋和Ｈｇ２＋为明显．该菌与苯胺降解代谢有关酶类的测定结果表明，该菌含有的邻苯二酚－２，３加双氧酶是诱导酶．     

一株海洋柴油降解菌的分离筛选鉴定及其生物表面活性剂特性分析

从中国浙江省舟山渔场油污染的海水和海洋沉积物中分离筛选产生物表面活性剂的柴油降解菌株。经富集培养、形态观察、测定单菌噬油斑、柴油降解率大小初筛到3株柴油降解菌。然后对初筛到的3株菌进行液滴坍塌实验、发酵液的表面张力、排油圈和乳化稳定性的大小测定进一步复筛,最终筛选出1株产生物表面活性剂的柴油降解菌,经18s rRNA鉴定为海洋解脂耶罗威亚酵母（Yarrowia lipolytica）。其柴油降解率为80%,发酵液液体表面张力可从73.4 mN/m降至23.56 mN/m,乳化效率E₂₄为60%。通过薄层色谱、傅里叶变换红外光谱、GC/MS鉴定,其产生的表面活性剂是由C₁₄、C₁₅β-羟基脂肪酸组成的脂肽。