分离海洋不动杆菌及其对石油烃降解性能研究

石油降解菌在石油污染生物修复技术中起到非常重要的作用。本研究分别以渤海湾油污区采集的水样,油样,水油泥混合样为材料富集分离石油降解菌,对其进行生理生化及分子生物学鉴定,并采用GC-MS测定烷烃、环烃、芳香烃等石油烃组分的变化。其中3株菌具有较高石油烃降解能力,16SrRNA序列分析表明该3株菌均与不动杆菌属（Acinetobacter）有99%序列相似性,可初步鉴定为不动杆菌属（Acinetobacter）。3株菌的石油烃降解能力依次为Tust-DM21＞Tust-DC12＞Tust-DW04,对原油成分的降解效果依次为烷烃＞芳香烃＞环烃。其中菌株Tust-DM21为一株高效石油烃降解菌,28℃于富集培养基培养10 d后,对烷烃（C10~C30）的降解率可达98%,对芳香烃和环烃的降解率达88%。研究表明,Tust-DM21菌株对烷烃,环烃,芳香烃都有较强的降解能力,是一株具有较好开发前景的石油降解菌。     

低温条件下硝基苯降解菌的分离及降解特性

低温条件下,从东北制药总厂的曝气池与氯霉素生产废水集水池污泥中分离、筛选得到6株以硝基苯为唯一碳源的降解菌,对6株菌进行降解能力研究得知,菌株cc-2为低温高效硝基苯降解菌。经形态特征和生理生化特征分析,初步鉴定属于不动杆菌属(Acinetobacter sp.)。菌株cc-2降解性能研究结果表明:菌株最佳条件为生长温度15℃、培养基pH值为7、摇床转速为140 r/min。最佳降解条件下,当硝基苯初始浓度为200 mg/L时菌株48h降解率达66.84%。外加葡萄糖和乙酸钠促进了菌株cc-2对硝基苯的生物降解,降解率分别为80.44%和78.57%,该菌株的生长降解性能研究为低温环境中硝基苯的修复提供了技术支撑。     

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.     

菲降解菌株的分离鉴定及特性研究

从某石化公司污水处理厂的活性污泥中分离到1株对菲具有降解活性的菌株AFF,该菌株能够以菲为唯一碳源和能源生长.根据形态特征及16S rRNA基因序列分析,该菌株初步鉴定为Burkholderia sp..在以菲为唯一碳源的培养基中,84 h内菌株对100 mg·L-1菲的降解率可达74.1%.通过液相-质谱联用及紫外可见分光光度法分析,推测该菌株代谢菲的途径为水杨酸途径.测定了降解途径中相关酶的活性,表明该菌株降解酶系中具有邻苯二酚2,3-双加氧酶活性,并证明该酶为诱导酶;确定了粗酶降解邻苯二酚的动力学常数:米氏常数Km为10.93μmol·L-1,最大反应速率Vmax为106.38μmol·min-1,说明酶与邻苯二酚的亲和力较大,邻苯二酚是该酶的良好底物.     

多氯联苯降解菌Pseudomonas sp. DN2的分离鉴定及其降解特性研究

从长期受PCBs污染的土壤中经富集培养筛选分离到1株能以联苯为唯一碳源和能源生长的革兰氏阴性细菌DN2.经形态观察、16S rDNA序列分析和系统发育分析初步鉴定为Pseudomonas sp..对其联苯双加氧酶的bphA1基因核心区和2,3-二羟基联苯-1,2-双加氧酶活力进行了分析测定,结果表明该菌具有降解联苯/PCBs的相关基因存在.利用GC-MS测试该菌的静息细胞降解PCBs的能力表明,该菌株可以在3 d内显著而高效地降解Aroclor 1242总量的67%,其对Aroclor 1242中各单体同系物的降解率在30%～90%之间.这对PCBs污染场地的原位生物修复具有实际应用意义.     

一株苯酚降解菌的分离和鉴定

:从工业废水中分离到一株高效降解苯酚的菌株PHEA 2 ,它能够在以苯酚为单一碳源和能源的无机培养基上生长 ,其最高苯酚降解率为 394nmol/ (mg·min) .经Biolog细菌自动鉴定仪鉴定 ,该菌株为醋酸钙不动杆菌 (Acinetobactercalcoaceti cus) .采用PCR扩增获得的该菌 16SrDNA片段 ,核苷酸序列分析结果表明 ,该菌株的 16SrDNA的核苷酸序列与醋酸钙不动杆菌同源性为 98% .在细菌系统发育分类学上 ,PHEA 2归属变形细菌γ亚类、不动杆菌属、醋酸钙不动杆菌 .     

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.     

Pentachlorophenol degradation by Pseudomonas stutzeri CL7 in the secondary sludge of pulp and paper mill

A pentachlorophenol (PCP) mineralizing bacterium was isolated from the secondary sludge of pulp and paper mill and identified as Pseudomonas stutzeri strain CL7. This isolate used PCP as its sole source of carbon and energy and was capable of degrading this compound as indicated by stoichiometric release of chloride and biomass formation. P. stutzeri (CL7) was able to mineralize a high concentration of PCP (600 mg/L) than any previously reported Pseudomonad with PCP as sole carbon source. As the concentration of PCP increased from 50 to 600 mg/L, the reduction in the cell growth was observed and the PCP degradation was more than 90% in all studied concentrations. This isolate was able to remove 66.8% of PCP from the secondary sludge of pulp and paper mill when supplemented with 100 mg/L of PCP and grown for two weeks. This study showed that the removal e ciency of PCP by CL7 was found to be very e ective and can be used in PCP remediation of pulp paper mill waste in the environment.     

1株咔唑降解菌的分离、鉴定及其降解特性研究

利用微生物分离技术,从土壤中分离到1株咔唑降解菌KH-6.菌株KH-6能够以咔唑为唯一的碳源和能源良好地生长,同时还可以利用3-甲基咔唑、4-羟基咔唑和2,2′-二羟基联苯生长.经过对其形态特征、生理生化、以及16S rDNA序列分析,初步鉴定为黄杆菌属细菌.该菌株对咔唑的降解性是通过在液体培养基内菌体的增加及底物的减少来证实的,其利用咔唑生长的最适温度和pH分别为30℃和7.5.静止细胞反应试验表明,菌株KH-6除了咔唑以外还可降解其它杂环化合物.咔唑污染土壤中的生物降解实验表明,在灭菌土壤中90%的咔唑被降解,菌株KH-6对土壤中的咔唑有很好的生物降解效果.     

环境激素对叔辛基酚降解菌的筛选、鉴定及降解特性

为了研究环境激素4-t-OP(对叔辛基酚)的生物降解,从扬州市汤汪生活污水处理厂二沉池污泥中筛选得到1株能以4-t-OP为唯一碳源进行生长的降解菌株,标示为TW30,16S rRNA测试其为不动杆菌属(Acinetobacter sp.),通过摇瓶试验测试其降解活性.结果表明:在40℃、初始pH为6.0、ρ(4-t-OP)为5 mg/L的无机盐培养基中,5 d后降解率可达99.03%;降解过程满足一级反应模型,降解速率常数(k)为0.875 d-1,半衰期(t1/2)为0.8 d.这说明TW30是一株高效的4-t-OP降解菌.此外,培养温度的升高和额外Ca2+、Mn2+的加入可以提高TW30降解4-t-OP的效率,而在5～25 mg/L范围内提高初始ρ(4-t-OP)以及额外加入磷酸盐、NH4+、Mg2+、Fe2+、Na+、Zn2+、Cu2+等无机盐和葡萄糖、CH3COO-等碳源则会降低降解率.      

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．     

DMF降解菌MBYD-1的分离、鉴定及降解特性

从农药厂污水处理池的活性污泥中分离得到一株能以二甲基甲酰胺(DMF)为唯一碳、氮源生长的菌株MBYD-1.经形态观察、生理生化实验及16S rRNA基因序列分析,将菌株初步鉴定为假单胞菌属 (Pseudomonas sp.).菌株MBYD-1降解DMF的最适条件为:pH 7.0,30℃.当DMF浓度为400mg/L,接种量为2%时,菌株能在72h内将DMF完全矿化.底物广谱性实验表明,菌株MBYD-1能在甲酸盐和甲醛中很好地生长,在甲酰胺中的生长情况优于二甲胺.     

接合子细胞强化对生物反应器降解2，4-D效应研究

研究了接合性质粒pJP4在2株纯菌E．coliDH5α、Alcaligenes sp．及好氧颗粒污泥混合菌群中的水平转移情况,并以获得pJP4质粒的接合子Alcaligenes sp．：：pJP4为强化菌种,考察了接合子细胞强化对好氧颗粒反应器和生物膜反应器中难降解有机物2,4-二氯苯氧乙酸（2,4-D）的去除效应．结...     

苯并[a]芘降解菌SL-1的筛选、鉴定及其降解特性

从土壤中筛选出一株以苯并[a]芘为唯一碳源的高效降解菌SL-1,经形态观察、生理生化分析以及16SrDNA基因序列分析,该菌株鉴定为假单胞菌（Pseudomonassp．）。28℃震荡培养10d后,菌株SL-1对5mg·L^-1苯并[a]芘的降解率为35．7％;同时测定该降解菌的生长曲线、降解率变化曲线、最佳培养条件;土壤修复试验表明,42d时,降解菌SL-1对苯并芘的降解率达到76．3％。     

Isolation and characteristics of a novel biphenyl-degrading bacterial strain, Dyella ginsengisoli LA-4

A novel biphenyl-degrading bacterial strain LA-4 was isolated from activated sludge. It was identified as Dyella ginsengisoli according to phylogenetic similarity of 16S rRNA gene sequence. This isolate could utilize biphenyl as sole source of carbon and energy, which degraded over 95 mg/L biphenyl within 36 h. The major metabolites formed from biphenyl, such as 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid (HOPDA) and benzoic acid, were identified by LC-MS. The crude cell extract of strain LA-4 exhibited the activity of 2,3-dihydroxybiphenyl 1,2-dioxygenase (2,3-DHBD) and the kinetic parameters were Km= 26.48 μmol/L and Vmax= 8.12 μmol/mg protein. A conserved region of the biphenyl dioxygenase gene bphA1 of strain LA-4 was amplified by PCR and confirmed by DNA sequencing.     

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.     

Rhodococcus sp. Ns对硝基苯酚的好氧生物降解

通过驯化富集培养,从红树林底泥中分离出6株硝基苯酚降解菌,其中Rhodococcus sp. Ns为对硝基苯酚（PNP）与邻硝基苯酚（ONP）的高效降解菌.在好氧条件下该菌可以耐受小于1.8 mmol/L的PNP,能够利用PNP和ONP为唯一碳源、能源和氮源生长并将其完全矿化.研究了Rhodococcus sp. Ns在不同pH、盐度与浓度范围下,PNP的降解特性并探讨了该菌降解PNP的途径.实验得出该菌在盐度<5‰、 pH>5的条件下能较快生长,1.5 mmol/L的PNP在96h内被完全降解,并检测到至少2种中间产物4-硝基儿茶酚(4-nitrocatechol)和1,2,4-苯三酚 (1,2,4-benzenetriol).红树林底泥中固有的细菌对PNP和ONP具有高效降解作用.     

菌株EA41对醋酸酯类化合物的生物降解

利用醋酸乙酯为能源和碳源从土壤中分离到一株菌。在生理生化试验的基础上,利用16S rDNA的方法鉴定这株菌为假单胞菌属细菌。底物特异性试验表明,假单胞菌EA41菌株可代谢多种醋酸酯类化合物。休眠细胞反应试验表明,这株菌的水解酶能水解口香糖的主要成分聚醋酸乙烯酯产生醋酸。     

17α-乙炔基雌二醇降解菌的分离鉴定及降解特性

从避孕药生产厂废水处理站的活性污泥中驯化、分离到1株能够以17α-乙炔基雌二醇(17-αethynylestradiol,EE2)为唯一碳源和能源生长的菌株JCR5.经过对其形态特征、生理生化以及16S rDNA序列分析,该菌株为鞘氨醇杆菌属(Sphingobacterium sp.).研究表明,菌株JCR5利用EE2生长的适宜温度为25～40℃,培养基初始pH为7～9.金属离子Ni²⁺、Mn²⁺、Cu²⁺、Fe³⁺能够促进菌株的生长,而Zn²⁺、Ag⁺、Pb²⁺、Ca²⁺和Al³⁺离子对菌株的生长具有不同程度的抑制作用.菌株JCR5在10d内对初始底物浓度为30mg·L^-1EE2的降解率可达到87%.     

好氧颗粒污泥及其高效菌株降解苯胺的试验研究

以苯胺为唯一碳源和氮源,培养降解高浓度苯胺废水的好氧颗粒污泥,该体系对苯胺废水的最高耐受浓度高达6 000mg/L.通过分离纯化,从颗粒污泥体系中获得2株具有不同降解特征的苯胺降解菌adx1和adx3,菌株adx1在降解速率上具有明显优势,而菌株adx3对苯胺的最高耐受浓度高于adx1.上述菌株在降解苯胺过程中均遵循Haldane动力学模型,菌株adx1和adx3的最大比较降解速率分别为0.924 g/(g.h)和0.645 g/(g.h),比生长速率分别为0.487 g/(g.h)和0.440 g/(g.h).16S rDNA测序结果表明adx1和adx3分别属于Pseudomonas和Achromobacter属,与好氧颗粒污泥PCR-DGGE指纹图条带1和4测序结果一致,表明上述菌株分别为好氧颗粒化体系中优势菌群之一.