一株产脂肽类表面活性剂的碱性Dietzia菌及特性研究

【目的】筛选降解性能良好的产生物表面活性剂的菌株,对其进行分类学鉴定,确定所产表面活性剂物质并对各影响因素进行评价。【方法】利用液体石蜡为底物筛选降解性能良好的产生物表面活性剂菌株,通过形态特征观察、生理生化测定、16S rRNA基因序列分析等实验确定菌株的分类地位。通过排油圈活性、表面张力值、薄层层析等方法确定生物表面活性剂的性质,分析碳、氮源和温度、pH、盐浓度各因素对菌株产生物表面活性剂的影响。【结果】从大连新港采集的样品中分离得到一株产表面活性剂的嗜碱菌株3372,经分类鉴定表明其是Dietzia cercidiphylli的新菌株。嗜碱菌3372发酵液粗提物的排油直径为6.1 cm,表面张力可从67.62 mN/m降到32.95 mN/m,经薄层层析分析,初步鉴定为脂肽类表面活性剂。综合各因素对发酵液表面活性的影响,菌株3372在pH为9.0、适盐浓度为3%的培养基中,经30°C培养可将发酵液表面张力值降到最低。【结论】嗜碱菌3372是脂肽类生物表面活性剂产生菌的新成员,其在高盐碱条件下产生表面活性剂的特性在工业应用上有一定的潜力。     

苯扎贝特降解菌的筛选及降解特性

【目的】药品苯扎贝特在水环境中频繁检测出,对环境的潜在危害不容忽视。我们筛选分离降解苯扎贝特的细菌,并研究其降解特性。【方法】根据分离菌株的细胞形态结构、生理生化特征及其16S rRNA基因序列分析鉴定降解菌,高效液相色谱法测定苯扎贝特,以判定该菌株的降解能力。【结果】分离菌株B-31属恶臭假单胞菌(Pseudomonas putida),降解机制是共代谢。降解最佳条件为30℃、pH7。此条件下,以1%甲醇为初级基质,30mg/L苯扎贝特的5日降解率为48%。当分别以5g/L葡萄糖、蛋白胨、酵母粉为初级基质时,可使降解率提高到61%,、72.6%、76.67%。【结论】这是国内首次报道恶臭假单胞菌可以通过共代谢降解苯扎贝特,该研究为利用细菌发酵消除水环境中苯扎贝特污染提供基础。     

一株糖脂表面活性剂产生菌的筛选及干酪根降解

【目的】从油页岩环境中筛选可降解油页岩干酪根的产生物表面活性剂菌株。【方法】从抚顺油页岩矿废水样品中用血平板法初筛,排油圈法、乳化法和表面张力法复筛,获得产生物表面活性剂菌株。对目标菌株进行生理生化鉴定、16S r RNA基因序列和系统发育分析,用薄层色谱鉴定其发酵液表面活性成分,优化产表面活性剂的培养条件,初步考察其对油页岩干酪根的降解能力。【结果】筛选到一株产糖脂表面活性剂菌株B-1,初步鉴定为Pseudomonas sp.,该菌株有良好的排油和乳化能力以及较低的表面张力,可利用烷烃、不饱和脂肪酸和糖类作为碳源。在30-34°C范围内添加0.3%Na Cl的葡萄糖培养基(p H 7.0)中该菌生长旺盛,发酵液表面张力最低为27 m N/m。菌株B-1在添加一定量葡萄糖的无机盐培养基中作用30 d后对干酪根的降解率为2.85%,高于不添加葡萄糖无机盐培养基对照组的降解率(1.04%)。【结论】菌株B-1是一株性能良好的产糖脂表面活性剂细菌,有降解干酪根的潜力。     

一株苯系物降解真菌筛选及降解特性

采用逐量分批驯化的方法以污水处理厂污泥作为菌源,苯、甲苯、二甲苯为唯一碳源,驯化、分离、筛选能够有效降解苯系物的真菌,命名为B1。采用单因素以及正交实验方法并对真菌降解环境影响因素及降解效率进行了测定和研究。结果表明:真菌B1对苯系物降解的最佳条件为C:N=5:1,pH5,温度30℃,菌种接种量为5.5ml(50ml培养基)。采用GC对初始液相浓度0~90mg/L范围内的苯系物降解效果进行测定,未发现苯系物对真菌降解活性产生抑制作用。真菌对苯系物的降解效率为:甲苯>苯>二甲苯,最高降解效率分别达到87.39%,85.21%,81.47%。混合物降解效果略高于单一底物的降解效果。     

一株产甘露糖赤藓糖醇脂菌株的筛选及其产物分析

【目的】解决石油长链烃类物质引起的环境污染问题,筛选可以高效降解石油烃的产糖脂类生物表面活性剂菌株。【方法】采用血平板、油平板法,从葡萄皮表面分离到6株产糖脂类的真菌,比较各菌株的排油性能,通过PCR扩增合成糖脂类表面活性剂的关键基因,筛选到一株具有emtl序列的真菌K6。经形态学、生理生化测定和分子系统发育分析(5.8S,ITS1,ITS2)对菌株进行鉴定,而且通过TLC和HPLC分析该菌株的代谢产物。【结果】经鉴定,该菌为Pseudozyma churashimaensis,可产甘露糖赤藓糖醇脂。石油烃降解实验表明,菌株K6具有很强的乳化性能和降解石油烃的能力,其石油烃降解率可达70.17%。【结论】菌株K6具有产生物表面活性剂和降解长链石油烃类的能力,其对石油污染环境的生物修复具有重要的现实意义。     

一株高效降解芘的细菌分离、鉴定及其降解效果

摘要：【目的】获得高效降解高分子量多环芳烃的细菌,并研究其对多环芳烃的降解能力。【方法】利用富集培养和芘升华平板方法,从焦化厂污染土壤中分离多环芳烃降解细菌,对分离菌株通过形态特征、16S rRNA基因和gyrb基因序列相似性分析进行鉴定,并研究该菌对高分子量多环芳烃（HMW-PAHs）的降解效果。【结果】筛选到一株能以芘、苯并蒽、屈、苯并芘、茚并芘、苯并苝、荧恩为碳源和能源生长并降解这些底物的菌株HBS1,该菌株的16S rRNA基因和gyrb基因序列与Gordonia amicalis的相应基因的相似     

降解尿酸乳酸菌复合菌系的筛选与菌株组合提升降解效果

【背景】高尿酸症由血液中尿酸含量明显升高而导致,利用乳酸菌对人体的益生作用缓解高尿酸血症越来越受到关注。【目的】获得具有降解尿酸能力的乳酸菌复合菌系与纯培养菌株。【方法】以泡菜为样品来源,以尿酸为底物,采用MRS培养基筛选降解尿酸的乳酸菌复合菌系,通过高效液相色谱法测定复合菌系对尿酸的降解能力。【结果】得到一组乳酸菌复合菌系,当培养温度为37 °C、pH值为6.20、静置培养72 h后复合菌系对尿酸的降解率为12.08%;通过优化培养条件,当该菌系在以牛肉膏为单一氮源、初始pH值为5.00、温度为35 °C的条件下培养72 h,尿酸降解率上升至17.19%,降解率比优化前提高了42.3%;从该菌系中分离出两株具有尿酸降解能力的菌株UA-1与UA-2,它们的尿酸降解率分别为10.85%和8.65%;通过形态学观察和16S rRNA基因序列分析,经鉴定两株菌均为布氏乳杆菌(Lactobacillus buchneri)。将两株单菌组合降解尿酸试验发现,UA-1与UA-2比例为2:1的尿酸降解率为20.2%,比原复合菌系的降解能力提高了67.22%。【结论】研究证明了乳酸菌复合菌系对尿酸的降解能力优于单个菌株,为后续利用乳酸菌复合菌系应用提供了数据支持。     

反应器进水管生物膜中喹啉降解菌的分离鉴定及降解特性

【背景】喹啉是一类高毒、致癌且难降解的含氮杂环化合物,本实验室建立了一个长期高效运行的反硝化喹啉降解生物反应器。【目的】从反应器进水管富集的生物膜中筛选有氧条件下降解喹啉的菌株。【方法】通过以喹啉为唯一碳源的培养基来富集、分离、纯化菌株;利用16S rRNA基因的序列分析鉴定分离株的系统发育地位;比较不同pH及温度条件下菌株的喹啉降解特性。【结果】经鉴定,4株分离物Q1、Q3、Q7和Q8分别属于Sphingobium、Massilia、Rhodococcus和Dyadobacter属。降解实验表明,以上菌株均能在48 h内实现50 mg/L喹啉的完全去除,但各自表现出不同的降解特性,其中Q1、Q3和Q8在降解过程中都检测到了喹啉降解产物2-羟基喹啉的积累。降解喹啉的Sphingobium、Massilia和Dyadobacter属菌株尚未见报道。【结论】从喹啉降解生物反应器的进水管内分离的4株喹啉降解菌可为设计处理含喹啉工业废水的反应器提供新菌种资源,对于完善喹啉生物降解机理研究具有实际意义。     

炔草酯降解菌Sphingopyxis sp. WP68的分离鉴定与降解特性

【背景】炔草酯可以高效防除麦田恶性杂草,但炔草酯的生产和使用也对环境造成了破坏,对动物和人类健康造成了威胁。【目的】分离筛选炔草酯高效降解菌株,研究其降解特性,为炔草酯污染生物修复提供优良菌种资源。【方法】采集农药厂活性污泥样品,通过富集培养和含有炔草酯的LB培养基进行炔草酯降解菌株的分离,通过形态和生理生化特性以及16S rRNA基因序列分析确定其分类学地位,通过单因素试验从温度、pH、接种量和底物浓度等方面考察菌株对炔草酯的降解特性,并利用UPLC-MS分析降解产物。【结果】筛选出一株炔草酯高效降解菌株WP68,经鉴定为鞘氨醇盒菌(Sphingopyxis sp.),该菌株在37°C和pH值为8.0时,10 h内可将200 mg/L的炔草酯降解98.26%。利用UPLC-MS鉴定菌株WP68降解炔草酯的产物为炔草酸。确定了该菌株降解炔草酯的最适温度、pH值、接种量、底物浓度分别是37°C、8.0、5%、200mg/L。菌株WP68还能降解氰氟草酯和精喹禾灵。【结论】Sphingopyxis sp. WP68对炔草酯有较强的降解能力和较高耐受性,在炔草酯污染土壤修复中具有潜在的应用前景。     

一株高效DEHP降解菌的分离、鉴定及其降解特性

【目的】分离得到高效的邻苯二甲酸二乙基己基酯(DEHP)降解菌。【方法】采用富集培养法筛选分离菌株,并对菌株进行驯化;通过PCR扩增得到其16S rRNA和gyrB基因序列,进行同源序列分析及分子系统发育树的构建,同时结合形态学观察和生理生化实验对菌株进行初步鉴定;采用高效液相色谱(HPLC)分析菌株对DEHP的降解特性。【结果】分离得到一株能以DEHP为唯一碳源和能源生长的菌株,命名为HS-NH1,初步鉴定其为戈登氏菌(Gordoniasp.)。菌株HS-NH1最适的生长和降解条件为30°C、pH 7.0,在此条件下,该菌株60 h内能够将浓度为500 mg/L的DEHP降解90%以上。高效液相色谱(HPLC)分析表明,菌株HS-NH1在降解DEHP过程中产生了一种重要的中间代谢产物——邻苯二甲酸。底物广谱性试验证明,菌株HS-NH1能够有效地利用多种常见的邻苯二甲酸酯(PAEs)与芳香族衍生物。【结论】筛选得到了一株DEHP降解菌Gordonia sp.HS-NH1,该菌降解效率高,具有良好的底物广谱性,在邻苯二甲酸酯类化合物的污染治理中将会有一定的应用潜力。     

Biodegradation and biotransformation of groundwater pollutant mixtures by Mycobacterium vaccae.

Mycobacterium vaccae can catabolize a number of major groundwater pollutants. When added singly, acetone, cyclohexane, styrene, benzene, ethylbenzene, propylbenzene, dioxane, and 1,2-dichloroethylene can be catabolized by M. vaccae. Catabolism of a number of these chemicals was monitored by gas-chromatographic analysis. Gas-chromatographic analysis indicated that the products of benzene degradation are phenol and hydroquinone. The products of chlorobenzene and ethylbenzene degradation are 4-chlorophenol and 4-ethylphenol. The extent that some compounds were catabolized when present as mixtures was also investigated. When toluene and benzene were present concomitantly, toluene was catabolized and benzene oxidation was delayed. Although toluene promoted the degradation of styrene, a lower rate of toluene degradation occurred when styrene was present. Both 4-chlorophenol and 4-ethylphenol had an antagonistic effect on the ability of M. vaccae to degrade other aromatic compounds. Studies with [14C]benzene indicated that M. vaccae can mineralize small amounts of this compound. These results suggest that components in mixtures may have a positive or a negative effect on the rates of biodegradation of other pollutants.     

The conversion of BTEX compounds by single and defined mixed cultures to medium-chain-length polyhydroxyalkanoate

Here, we report the use of petrochemical aromatic hydrocarbons as a feedstock for the biotechnological conversion into valuable biodegradable plastic polymers-polyhydroxyalkanoates (PHAs). We assessed the ability of the known Pseudomonas putida species that are able to utilize benzene, toluene, ethylbenzene, p-xylene (BTEX) compounds as a sole carbon and energy source for their ability to produce PHA from the single substrates. P. putida F1 is able to accumulate medium-chain-length (mcl) PHA when supplied with toluene, benzene, or ethylbenzene. P. putida mt-2 accumulates mcl-PHA when supplied with toluene or p-xylene. The highest level of PHA accumulated by cultures in shake flask was 26% cell dry weight for P. putida mt-2 supplied with p-xylene. A synthetic mixture of benzene, toluene, ethylbenzene, p-xylene, and styrene (BTEXS) which mimics the aromatic fraction of mixed plastic pyrolysis oil was supplied to a defined mixed culture of P. putida F1, mt-2, and CA-3 in the shake flasks and fermentation experiments. PHA was accumulated to 24% and to 36% of the cell dry weight of the shake flask and fermentation grown cultures respectively. In addition a three-fold higher cell density was achieved with the mixed culture grown in the bioreactor compared to shake flask experiments. A run in the 5-l fermentor resulted in the utilization of 59.6 g (67.5 ml) of the BTEXS mixture and the production of 6 g of mcl-PHA. The monomer composition of PHA accumulated by the mixed culture was the same as that accumulated by single strains supplied with single substrates with 3-hydroxydecanoic acid occurring as the predominant monomer. The purified polymer was partially crystalline with an average molecular weight of 86.9 kDa. It has a thermal degradation temperature of 350 degrees C and a glass transition temperature of -48.5 degrees C.     

Surfactant-enhanced biodegradation of high molecular weight polycyclic aromatic hydrocarbons by stenotrophomonas maltophilia

The objectives of this study were to isolate and evaluate microorganisms with the ability to degrade high molecular weight polycyclic aromatic hydrocarbons (PAHs) in the presence of synthetic surfactants. Stenotrophomonas maltophilia VUN 10,010, isolated from PAH-contaminated soil, utilized pyrene as a sole carbon and energy source and also degraded other high molecular weight PAHs containing up to seven benzene rings. Various synthetic surfactants were tested for their ability to improve the PAH degradation rate of strain VUN 10,010. Anionic and cationic surfactants were highly toxic to this strain, and the Tween series was used as a growth substrate. Five nonionic surfactants (Brij 35, Igepal CA-630, Triton X-100, Tergitol NP-10, and Tyloxapol) were not utilized by, and were less toxic to, strain VUN 10,010. MSR and log Km values were determined for fluoranthene, pyrene, and benzo[a]pyrene in the presence of these nonionic surfactants and their apparent solubility was increased by a minimum of 250-fold in the presence of 10 g L-1 of all surfactants. The rate of pyrene degradation by strain VUN 10,010 was enhanced by the addition of four of the nonionic surfactants (5-10 g L-1); however, 5 g L-1 Igepal CA-630 inhibited pyrene degradation and microbial growth. The specific growth rate of VUN 10,010 on pyrene was increased by 67% in the presence of 10 g L-1 Brij 35 or Tergitol NP-10. The addition of Brij 35 and Tergitol NP-10 to media containing a single high molecular weight PAH (four and five benzene rings) as the sole carbon source increased the maximum specific PAH degradation rate and decreased the lag period normally seen for PAH degradation. The addition of Tergitol NP-10 to VUN 10,010 cultures which contained a PAH mixture (three to seven benzene rings) substantially improved the overall degradation rate of each PAH and increased the specific growth rate of VUN 10,010 by 30%. Evaluation of the use of VUN 10,010 for degrading high molecular weight PAHs in leachates from surfactant-flushed, weathered, PAH-contaminated sites is warranted. Copyright 1998 John Wiley & Sons, Inc.     

Sphingobacterium bambusaue及其紫外诱变菌株的石油降解功能

【目的】研究Sphingobacterium bambusaue及其紫外诱变菌株的石油降解功能。【方法】紫外诱变后筛选石油降解高效菌株; 以不同石油浓度、pH值及盐浓度优化培养条件, 用重量法检测高效菌株石油降解率。【结果】发现菌株S. bambusaue在石油降解培养基中培养5 d的石油降解率为25.86%, UV诱变高效菌株IBFC2009-S3培养5 d的石油降解为42.85%, 比始发菌株提高65.7%; UV诱变菌株IBFC2009-S3的优化培养条件为石油浓度0.5 g/L、pH值7.0以及NaCl质量浓度为10 g/L, 其石油降解率可达50.51%。【结论】首次报道S. bambusaue具有石油降解功能; 紫外诱变获得的菌株S3的石油降解能力较强。     

Spore-forming, Desulfosporosinus-like sulphate-reducing bacteria from a shallow aquifer contaminated with gasoline

Previous studies on the geochemistry of a shallow unconfined aquifer contaminated with hydrocarbons suggested that the degradation of some hydrocarbons was linked to bacterial sulphate reduction. There was attenuation of naphthalene, 1,3,5-trimethylbenzene (TMB), toluene, p-xylene and ethylbenzene in the groundwater with concomitant loss of sulphate. Here, the recovery of eight strains of sulphate-reducing bacteria (SRB) from the contaminated site is reported. All were straight or curved rod-shaped cells which formed endospores. Amplification and sequencing of the 16S rDNA indicated that the strains were all sulphate reducers of the Gram-positive line of descent, and were most closely related to Desulfosporosinus (previously Desulfotomaculum) orientis DSM 8344 (97-98.9% sequence similarity). The strains clustered in three phylogenetic groups based on 16S rRNA sequences. Whole cell fatty acid compositions were similar to those of D. orientis DSM 8344, and were consistent with previous studies of fatty acids in soil and groundwater from the site. Microcosms containing groundwater from this aquifer indicated a role for sulphate reduction in the degradation of [ring-UL-14C]toluene, but not for the degradation of [UL-14C]benzene which could also be degraded by the microcosms. Adding one of the strains that was isolated from the groundwater (strain T2) to sulphate-enriched microcosms increased the rate of toluene degradation four- to 10-fold but had no effect on the rate of benzene degradation. The addition of molybdate, an inhibitor of sulphate reduction, to the groundwater samples decreased the rate of toluene mineralization. There was no evidence to support the mineralization of [UL-14C]benzene, [ring-UL-14C]toluene or unlabelled m-xylene, p-xylene, ethylbenzene, TMB or naphthalene by any of the strains in pure culture. Growth of all the strains was completely inhibited by 100 micromol l-1 TMB.     

一株纤维素降解菌的分离、鉴定及对 玉米秸秆的降解特性

[目的]获得高产纤维素酶细菌菌株,探讨以氨化预处理玉米秸秆为底物时的纤维素酶产酶特性及底物降解特性,探讨纤维素酶作用机理,提高玉米秸秆利用率.[方法]用LB培养基分离并纯化菌株,羧甲基纤维素钠培养基培养、刚果红染色进行初步筛选.考察氨化预处理对底物降解率、产酶能力的影响.通过形态特征观察及16S rRNA、Biolog鉴定菌株.[结果]分离到一株高效纤维素降解菌NH11,经鉴定为枯草芽孢杆菌(Bacillus subtilis). 30℃、发酵5d时,预处理前后玉米秸秆降解率分别为14.24％和24.73％.30℃、pH 7.2时,处理组CMC酶活力峰值处为153.84 U/mL,FPA酶活力为197.24 U/mL,比未处理组分别高出11.45％和10.59％.[结论]NH11具有较高的纤维素酶产酶能力,氨化预处理能够提高菌株对玉米秸秆的降解率.该菌株在秸秆堆肥、制作食用菌培养基和制取反刍动物粗饲料方面具有很高的应用价值.     

Biodegradation of benzene by halophilic and halotolerant bacteria under aerobic conditions

A highly enriched halophilic culture was established with benzene as the sole carbon source by using a brine soil obtained from an oil production facility in Oklahoma. The enrichment completely degraded benzene, toluene, ethylbenzene, and xylenes within 1 to 2 weeks. Also, [14C]benzene was converted to 14CO2, suggesting the culture's ability to mineralize benzene. Community structure analysis revealed that Marinobacter spp. were the dominant members of the enrichment.     

Stimulating the anaerobic degradation of aromatic hydrocarbons in contaminated sediments by providing an electrode as the electron acceptor

The possibility that electrodes might serve as an electron acceptor to simulate the degradation of aromatic hydrocarbons in anaerobic contaminated sediments was investigated. Initial studies with Geobacter metallireducens demonstrated that although toluene was rapidly adsorbed onto the graphite electrodes it was rapidly oxidized to carbon dioxide with the electrode serving as the sole electron acceptor. Providing graphite electrodes as an electron acceptor in hydrocarbon‐contaminated sediments significantly stimulated the removal of added toluene and benzene. Rates of toluene and benzene removal accelerated with continued additions of toluene and benzene. [¹⁴C]‐Toluene and [¹⁴C]‐benzene were quantitatively recovered as [¹⁴C]‐CO₂, demonstrating that even though the graphite adsorbed toluene and benzene they were degraded. Introducing an electrode as an electron acceptor also accelerated the loss of added naphthalene and [¹⁴C]‐naphthalene was converted to [¹⁴C]‐CO₂. The results suggest that graphite electrodes can serve as an electron acceptor for the degradation of aromatic hydrocarbon contaminants in sediments, co‐localizing the contaminants, the degradative organisms and the electron acceptor. Once in position, they provide a permanent, low‐maintenance source of electron acceptor. Thus, graphite electrodes may offer an attractive alternative for enhancing contaminant degradation in anoxic environments.     

可降解吡啶的全食副球菌B21-3的筛选鉴定及降解特性

【背景】吡啶作为一种难降解的有机污染物普遍存在于焦化、炼油、皮革和制药等行业的废水中,并对环境造成危害。【目的】治理废水中残留的有机污染物吡啶,筛选高效降解菌。【方法】采用富集培养和选择培养,以石家庄某污水处理厂的活性污泥为材料进行吡啶降解菌的筛选,通过形态特征、生理生化特性、(G+C)mol%测定及16S rRNA基因序列系统发育分析对筛选到的降解菌进行鉴定,并分析其对吡啶的降解特性。【结果】分离筛选到一株能以吡啶为唯一碳源和氮源生长代谢的降解菌B21-3,经鉴定该菌株为全食副球菌(Paracoccuspantotrophus)。菌株B21-3对吡啶的最适降解温度为32°C,最适降解pH为7.0,吡啶浓度为100mg/L时降解率为48.50%±0.02%;通过逐步提高吡啶初始浓度对菌株进行驯化,驯化后菌株可耐受较高浓度吡啶且吡啶降解率显著增加,吡啶浓度为100 mg/L时驯化后菌株B21-3对吡啶的降解率为90.26%±1.70%。驯化后菌株在含吡啶的无机盐平板上传代培养15代后,对吡啶的降解率为89.39%±2.03%。【结论】菌株B21-3具有较强的吡啶降解能力及降解稳定性,该菌株可作为吡啶污染水体生物修复的潜在资源。