Yeast and bacteria cell hydrophobicity and hydrocarbon biodegradation in the presence of natural surfactants: rhamnolipides and saponins

This study concerns the relation between hydrocarbon biodegradation in the presence of natural surfactants and cell hydrophobicity resulting from the use of these surfactants. The relative capabilities of two bacterial strains (Pseudomonas aeruginosa and Bacillus subtilis) and two yeast strains (Candida maltosa, Yarrowia lipolytica) were investigated. The selected microorganisms were tested separately and in combination in order to achieve the optimal degrading performance. The surface cell hydrophobicity of microorganisms and the degree of hydrocarbon biodegradation were measured. The microbial adhesion to the hydrocarbon (MATH) test was used to denote the surface cell hydrophobicity of the microbial species. The results indicate the correlation between the modification of the surface cell and the degree of hydrocarbon biodegradation; however results for bacteria differ from that obtained for yeast strains. Saponins, as the surfactant, was more effective than rhamnolipides during hydrocarbon biodegradation, though the concentration of this surfactant has no significant influence on the surface cell hydrophobicity.     

Nutrients Can Enhance the Abundance and Expression of Alkane Hydroxylase CYP153 Gene in the Rhizosphere of Ryegrass Planted in Hydrocarbon-Polluted Soil

Plant-bacteria partnership is a promising strategy for the remediation of soil and water polluted with hydrocarbons. However, the limitation of major nutrients (N, P and K) in soil affects the survival and metabolic activity of plant associated bacteria. The objective of this study was to explore the effects of nutrients on survival and metabolic activity of an alkane degrading rhizo-bacterium. Annual ryegrass (Lolium multiflorum) was grown in diesel-contaminated soil and inoculated with an alkane degrading bacterium, Pantoea sp. strain BTRH79, in greenhouse experiments. Two levels of nutrients were applied and plant growth, hydrocarbon removal, and gene abundance and expression were determined after 100 days of sowing of ryegrass. Results obtained from these experiments showed that the bacterial inoculation improved plant growth and hydrocarbon degradation and these were further enhanced by nutrients application. Maximum plant biomass production and hydrocarbon mineralization was observed by the combined use of inoculum and higher level of nutrients. The presence of nutrients in soil enhanced the colonization and metabolic activity of the inoculated bacterium in the rhizosphere. The abundance and expression of CYP153 gene in the rhizosphere of ryegrass was found to be directly associated with the level of applied nutrients. Enhanced hydrocarbon degradation was associated with the population of the inoculum bacterium, the abundance and expression of CYP153 gene in the rhizosphere of ryegrass. It is thus concluded that the combination between vegetation, inoculation with pollutant-degrading bacteria and nutrients amendment was an efficient approach to reduce hydrocarbon contamination.     

Isolation of alkane‐degrading bacteria from deep‐sea Mediterranean sediments

Aims: To isolate and identify alkane‐degrading bacteria from deep‐sea superficial sediments sampled at a north‐western Mediterranean station. Methods and Results: Sediments from the water/sediment interface at a 2400 m depth were sampled with a multicorer at the ANTARES site off the French Mediterranean coast and were promptly enriched with Maya crude oil as the sole source of carbon and energy. Alkane‐degrading bacteria belonging to the genera Alcanivorax, Pseudomonas, Marinobacter, Rhodococcus and Clavibacter‐like were isolated, indicating that the same groups were potentially involved in hydrocarbon biodegradation in deep sea as in coastal waters. Conclusions: These results confirm that members of Alcanivorax are important obligate alkane degraders in deep‐sea environments and coexist with other degrading bacteria inhabiting the deep‐subsurface sediment of the Mediterranean. Significance and Impact of the Study: The results suggest that the isolates obtained have potential applications in bioremediation strategies in deep‐sea environments and highlight the need to identify specific piezophilic hydrocarbon‐degrading bacteria (HCB) from these environments.     

Activity and viability of polycyclic aromatic hydrocarbon‐degrading Sphingomonas sp. LB126 in a DC‐electrical field typical for electrobioremediation measures

There has been growing interest in employing electro‐bioremediation, a hybrid technology of bioremediation and electrokinetics for the treatment of contaminated soil. Knowledge however on the effect of weak electrokinetic conditions on the activity and viability of pollutant‐degrading microorganisms is scarce. Here we present data about the influence of direct current (DC) on the membrane integrity, adenosine triphosphate (ATP) pools, physico‐chemical cell surface properties, degradation kinetics and culturability of fluorene‐degrading Sphingomonas sp. LB126. Flow cytometry was applied to quantify the uptake of propidium iodide (PI) and the membrane potential‐related fluorescence intensities (MPRFI) of individual cells within a population. Adenosine tri‐phosphate contents and fluorene biodegradation rates of bulk cultures were determined and expressed on a per cell basis. The cells' surface hydrophobicity and electric charge were assessed by contact angle and zeta potential measurements respectively. Relative to the control, DC‐exposed cells exhibited up to 60% elevated intracellular ATP levels and yet remained unaffected on all other levels of cellular integrity and functionality tested. Our data suggest that direct current (X = 1 V cm^?1; J = 10.2 mA cm^?2) as typically used for electrobioremediation measures has no negative effect on the activity of the polycyclic aromatic hydrocarbon (PAH)‐degrading soil microorganism, thereby filling a serious gap of the current knowledge of the electrobioremediation methodology.     

南海深海沉积物烷烃降解菌的富集分离与多样性初步分析

通过培养和非培养2种手段研究了南海沉积物中石油降解菌的多样性。通过烷烃富集培养,从2个站点不同深度的南海沉积物样品中富集筛选出48株深海细菌,其中27株对十六烷有降解能力。表面张力测定结果表明,4株降解菌同时具有较强的表面活性剂产生能力,2株Dietzia maris菌能使水的表面张力降至33mN/m左右,这是该种微生物产表面活性剂的首次报道。通过变性梯度凝胶电泳(DGGE)分析显示,南海沉积物富集物中的烷烃降解菌优势菌是芽孢杆菌,而且有多种。其中,Bacillus aquimaris在两个站点的7个样品的富集物中都是优势菌。此外,Sporosarcina,Halomona以及Brevibacterium属的细菌在不同样品中也表现为除Bacillus之外的优势菌。     

Optimization of <Emphasis Type="Italic">Enterobacter cloacae</Emphasis> (KU923381) for diesel oil degradation using response surface methodology (RSM)

Efficiency of Enterobacter cloacae KU923381 isolated from petroleum hydrocarbon contaminated soil was evaluated in batch culture and bioreactor mode. The isolate were screened for biofilm formation using qualitative and quantitative assays. Response surface methodology (RSM) was used to study the effect of pH, temperature, glucose concentration, and sodium chloride on diesel degradation. The predicted values for diesel oil degradation efficiency by the statistical designs are in a close agreement with experimental data (R ² = 99.66%). Degradation efficiency is increased by 36.78% at pH = 7, temperature = 35°C, glucose = 5%, and sodium chloride concentration = 5%. Under the optimized conditions, the experiments were performed for diesel oil degradation by gas chromatographic mass spectrometric analysis (GC-MS). GC-MS analysis confirmed that E. cloacae had highly degrade hexadecane, heptadecane, tridecane, and docosane by 99.71%, 99.23%, 99.66%, and 98.34% respectively. This study shows that rapid bioremoval of hydrocarbons in diesel oil is acheived by E. cloacae with abet of biofilm formation. The potential use of the biofilms for preparing trickling filters (gravel particles) for the degradation of hydrocarbons from petroleum wastes before their disposal in the open environment is highly suggested. This is the first successful attempt for artificially establishing petroleum hydrocarbon degrading bacterial biofilm on solid substrates in bioreactor.     

Empirical Models Estimating Carbon Dioxide Accumulation in Two Petroleum Hydrocarbon–Contaminated Soils

Bioremediation is a popular method in degrading diesel fuel contaminants from soil. Bioremediation can be enhanced by estimating the effect of important environmental parameters on microbial activity. Respirometry was used to develop empirical models describing the effects of temperature, moisture, nitrogen, and phosphorus concentration on microbial activity in a diesel-contaminated soil from Wyoming. Carbon dioxide (CO₂) data were analyzed using a base equation where its coefficient values were functions of each parameter. Two physiologically different groups of microorganisms were identified from the results under different operating temperatures. The empirical correlations were combined into one model and this model was tested against a hydrocarbon-contaminated soil collected from a site in Egypt with similar history of contamination. The predicted CO₂ evolution agreed well with the actual data obtained from the Egyptian soil samples, showing a sound predicting power of the empirical model for petroleum hydrocarbon biodegradation. Overall, the empirical correlations developed from the respirometric data provide a method to describe microbial activity in diesel-contaminated soils.     

Biodegradation of petroleum hydrocarbons in estuarine sediments: metal influence

In this work, the potential effect of metals, such as Cd, Cu and Pb, on the biodegradation of petroleum hydrocarbons in estuarine sediments was investigated under laboratory conditions. Sandy and muddy non-vegetated sediments were collected in the Lima River estuary (NW Portugal) and spiked with crude oil and each of the metals. Spiked sediments were left in the dark under constant shaking for 15 days, after which crude oil biodegradation was evaluated. To estimate microbial abundance, total cell counts were obtained by DAPI staining and microbial community structure was characterized by ARISA. Culturable hydrocarbon degraders were determined using a modified most probable number protocol. Total petroleum hydrocarbons concentrations were analysed by Fourier Transform Infrared Spectroscopy after their extraction by sonication, and metal contents were determined by atomic absorption spectrometry. The results obtained showed that microbial communities had the potential to degrade petroleum hydrocarbons, with a maximum of 32 % degradation obtained for sandy sediments. Both crude oil and metals changed the microbial community structure, being the higher effect observed for Cu. Also, among the studied metals, only Cu displayed measurable deleterious effect on the hydrocarbons degradation process, as shown by a decrease in the hydrocarbon degrading microorganisms abundance and in the hydrocarbon degradation rates. Both degradation potential and metal influence varied with sediment characteristics probably due to differences in contaminant bioavailability, a feature that should be taken into account in developing bioremediation strategies for co-contaminated estuarine sites.     

Structural characterization of the hydrocarbon degrading bacteria–oil interface: implications for bioremediation

Hydrocarbon degrading bacteria, enriched from an in situ bioremediation site in Long Valley, AZ emulsified and colonized the surface of waste engine oil. The application of a partial dehydration conventional embedding protocol for ultrathin-section transmission electron microscopy preserved the hydrocarbon degrading bacteria–surfactant–oil interface. Bacterial adsorption to oil occurred in association with a highly charged, amphipathic bacterial surfactant interface (25–50 nm thick). This biosurfactant completely encapsulated the emulsified oil droplets demonstrating that less than 1% surfactant (by volume) is required to emulsify waste hydrocarbon during or to promote biodegradation. Growth on oil appeared to occur by the uptake of tens of nm-sized droplets of emulsified oil.     

Comparative hydrocarbon utilization by hydrophobic and hydrophilic variants of Pseudomonas aeruginosa

Aims: To investigate hydrocarbon degradation by hydrophobic, hydrophilic and parental strains of Pseudomonas aeruginosa. Methods and Results: Partitioning of hydrocarbon‐degrading P. aeruginosa strain in a solvent/aqueous system yielded hydrophobic and hydrophilic fractions. Exhaustive partitioning of aqueous‐phase cells yielded the hydrophilic variants (L), while sequential fractionation of the hydrophobic phase cells yielded successive fractions exhibiting increasing cell‐surface hydrophobicity (CSH). In hydrocarbon adherence assays (bacterial attachment to hydrocarbon), L had a value of 20%, which increased from 61·7% in first hydrophobic fraction (H₁) to 72·2% in the third (H₃). Crude oil degradation by L was 70%, but increased from 82% in H₁ to 93% in H₃. L variant produced most exopolysaccharides and reduced surface tension from about 73 to 49 mN m^?1. Rhamnolipid production was highest in L, but was not detected in all crude oil cultures. Conclusions: Hydrophobic subpopulations of hydrocarbon‐degrading P. aeruginosa exhibited greater hydrocarbon‐utilizing ability than hydrophilic ones, or the parental strain. Significance and Impact of the Study: Results demonstrate that a population of P. aeruginosa consists of cells with different CSH which affect hydrocarbon utilization. This potentially provides the population with the capacity to utilize different hydrophobic substrates found in petroleum. Judicious selection of such hydrophobic subpopulations can enhance hydrocarbon pollution bioremediation.     

The Use of Conocarpus lancifolius Trees for the Remediation of Oil-Contaminated Soils

The role of the Conocarpus lancifolius tree in remediaitng oil-contaminated soil, which was bioremediated using conventional methods, was investigated. The selected tree was used to phytoremediate bioremediated oil-contaminated soil for three successive growing seasons. At the end of the phytoremediation experiment, 85.7% of measurable total petroleum hydrocarbon (TPH) was degraded in Conocarpus lancifolius rhizosphere, and the detectable concentrations of some poly aromatic hydrocarbons (PAHs) were less than 0.02 ppm. A number of hydrocarbon degrading microorganisms (HDMs) were isolated at 35°C under aerobic conditions, and were identified using 16S rRNA gene sequencing and fatty acid methyl ester (FAME) analysis. The efficiency of the isolated HDMs in degrading a mixture of hydrocarbon compounds (HC) was assessed. Among the bacterial isolates, Rhodococcus equi was distinguished from the other isolates because of its efficient degradation of some compounds in the HC mixture.

Samples were also collected from Conocarpus lancifolius vegetative parts and were analyzed for heavy metal and mineral accumulation. The results demonstrated that the Conocarpus lancifolius tree was able to uptake high levels of chromium (Cr), vanadium (V), and nickel (Ni) and accumulate them in the tree's roots. Additionally, Conocarpus trees tolerated high concentration and accumulated several metals in all plant tissues. These metals included aluminum (Al), calcium (Ca) and iron (Fe).     

Indigenous and enhanced mineralization of pyrene, benzo[a]pyrene, and carbazole in soils.

We studied the mineralization of pyrene, carbazole, and benzo[a]pyrene in soils obtained from three abandoned coal gasification plants in southern Illinois. The soils had different histories of past exposure to hydrocarbon contamination and different amounts of total organic carbon, microbial biomass, and microbial activity. Mineralization was measured by using serum bottle radiorespirometry. The levels of indigenous mineralization of 14C-labeled compounds ranged from 10 to 48% for pyrene, from undetectable to 46% for carbazole, and from undetectable to 25% for benzo[a]pyrene following long-term (greater than 180-day) incubations. Pyrene and carbazole were degraded with short or no lag periods in all soils, but benzo[a]pyrene mineralization occurred after a 28-day lag period. Mineralization was not dependent on high levels of microbial biomass and activity in the soils. Bacterial cultures that were capable of degrading pyrene and carbazole were isolated by enrichment, grown in pure culture, and reintroduced into soils. Reintroduction of a pyrene-degrading bacterium enhanced mineralization to a level of 55% within 2 days, compared with a level of 1% for the indigenous population. The carbazole degrader enhanced mineralization to a level of 45% after 7 days in a soil that showed little indigenous carbazole mineralization. The pyrene and carbazole degraders which we isolated were identified as a Mycobacterium sp. and a Xanthamonas sp., respectively. Our results indicated that mineralization of aromatic hydrocarbons can be significantly enhanced by reintroducing isolated polycyclic aromatic hydrocarbon-degrading bacteria.     

Surfactant enhances biodegradation of hydrocarbons: Microcosm and field study

The purpose of the present study was to provide new methods that would increase the rates of biodegradation of petroleum hydrocarbons in soil, thus reducing the time required to achieve a satisfactory level of residual hydrocarbon in an ex situ bioremediation. Results of laboratory studies on several techniques were used to guide our implementation of these methods in controlled field studies. Soils contaminated with nonvolatile hydrocarbons were treated with various combinations of (1) an anionic surfactant guanidinium cocoate (CGS), (2) a consortium of hydrocarbon‐degrading microorganisms, (3) a slow‐release form of nitrogen:urea, and (4) the bulking agent vermiculite. Laboratory results describing the activity of CGS have been presented previously (Jain et al., 1992). The amount and rate of hydrocarbon loss in treated soil was compared with hydrocarbon lost in soil that received no amendment other than water (water only). We also used a sheen screen method (Nelson et al., 1995), to assess the effectiveness of our field application of microorganisms.     

Indigenous and enhanced mineralization of pyrene, benzo[a]pyrene, and carbazole in soils.

We studied the mineralization of pyrene, carbazole, and benzo[a]pyrene in soils obtained from three abandoned coal gasification plants in southern Illinois. The soils had different histories of past exposure to hydrocarbon contamination and different amounts of total organic carbon, microbial biomass, and microbial activity. Mineralization was measured by using serum bottle radiorespirometry. The levels of indigenous mineralization of 14C-labeled compounds ranged from 10 to 48% for pyrene, from undetectable to 46% for carbazole, and from undetectable to 25% for benzo[a]pyrene following long-term (greater than 180-day) incubations. Pyrene and carbazole were degraded with short or no lag periods in all soils, but benzo[a]pyrene mineralization occurred after a 28-day lag period. Mineralization was not dependent on high levels of microbial biomass and activity in the soils. Bacterial cultures that were capable of degrading pyrene and carbazole were isolated by enrichment, grown in pure culture, and reintroduced into soils. Reintroduction of a pyrene-degrading bacterium enhanced mineralization to a level of 55% within 2 days, compared with a level of 1% for the indigenous population. The carbazole degrader enhanced mineralization to a level of 45% after 7 days in a soil that showed little indigenous carbazole mineralization. The pyrene and carbazole degraders which we isolated were identified as a Mycobacterium sp. and a Xanthamonas sp., respectively. Our results indicated that mineralization of aromatic hydrocarbons can be significantly enhanced by reintroducing isolated polycyclic aromatic hydrocarbon-degrading bacteria.     

中国东海和南海海域可培养烃类降解细菌的筛选及功能

【背景】海洋环境中蕴藏着丰富的微生物资源,其种类繁多而且功能多样,在驱动物质循环及能量流动等方面起着重要的作用。目前,海洋中烷烃化合物降解菌的分离筛选和降解功能研究已有文献报道;但是对海洋中尤其是我国东海和南海海域,具有降解芳香烃类化合物功能的菌株分离筛选及其多样性研究鲜有报道。【目的】分离筛选我国东海和南海海域具有烃类降解能力的可培养菌株,并对其降解功能和多样性进行初步研究。【方法】分别从东海和南海海底沉积物样品中筛选菌株,选择不同的烃类化合物为菌株筛选的唯一碳源,采用梯度稀释和平板划线法分离纯化得到单菌落,并利用相应烃类为唯一碳源进行生长验证获得该化合物降解菌。【结果】以肉桂酸、碱木素、十六烷等12种烃类化合物为唯一碳源,从样品中共分离到63株具有烃类化合物降解能力的菌株,分别属于3个门4个纲8个目10个属,主要为红球菌属(Rhodococcus)、不动杆菌属(Acinetobacter)、弧菌属(Vibrio)、盐单胞菌属(Halomonas)、假单胞菌属(Pseudomonas)。两大海域优势降解菌差别较大,其中东海沉积物降解菌株主要为不动杆菌属(Acinetobacter),而南海沉积物降解菌株主要为红球菌属(Rhodococcus)。【结论】我国东海和南海海域蕴藏着丰富的烃类化合物降解菌株资源,两大海域优势降解菌种类存在明显差异,这将为我国未来可能的海洋环境石油污染的微生物治理储备菌种资源。     

微贮用植物乳杆菌固定化菌剂的制备

乳酸菌与纤维素降解菌因其可防止微贮饲料酸败、增加秸秆饲料的营养价值等优点,在秸秆微贮过程中起重要作用。但由于乳酸菌的繁殖会抑制纤维素降解菌的活性,如何实现微贮过程中两种微生物分时发挥功能是解决上述问题的关键。文中利用固定化技术将乳酸菌制备成含有玉米秸秆粉的固定化菌剂以达到缓释的目的。首先制作固定化空白小球得出复合固定化载体成球的最佳浓度,利用玉米芯吸附植物乳杆菌S1得到复合固定化载体,以对S1的包埋率、成球效果等为指标,通过对比两种固定化方法 (包埋法与包埋-交联法),得到固定化植物乳杆菌S1的最佳条件。研究表明,使用6%PVA+0.4%SA+0.3%CMC-Na进行包埋-交联时成球效果最好,使用1.2%SA+0.5%CMC-Na进行直接包埋时成球效果最好。通过对比5种固定化工艺,将1.2%SA+0.5%CMC-Na和吸附玉米粉组成的固定化载体混合物逐滴滴入4%氯化钙中直接包埋24 h得到的固定化小球其机械强度以及包埋率均优于其他工艺。因此,利用玉米芯吸附-海藻酸钠包埋的方法可以有效提高植物乳杆菌包埋效率,为使用固定化技术制备微贮饲料菌剂奠定基础。     

A complementary approach to identifying and assessing the remediation potential of hydrocarbonoclastic bacteria

The isolation and assessment of hydrocarbonoclastic bacteria often represents a key strategy in the bioremediation of hydrocarbon-contaminated sites. However the isolation and assessment of such bacteria is often a lengthy and expensive procedure. The aim of this study was to identify potential isolates for use in the remediation of hydrocarbon contaminated sites using a combination of selective isolation plating, the Biolog system and subsequent multivariate analyses. The use of weathered oil as the main C source restricted the number of isolates growing to 5×10(2)CFUg soil(-1). Isolates (n=96) were then assessed individually using Biolog MT2 plates with seven different hydrocarbons (dodecane, tridecane, hexadecane, octadecane, eicosane, naphthalene and phenanthrene). The results indicated that all isolates were able to grow on at least one hydrocarbon from the seven chosen. This confirmed that the isolation media developed was selective in isolating hydrocarbonoclastic bacteria only. Cluster analysis of Biolog data separated the isolates into two discrete clusters with cluster 2 identifying hydrocarbonoclastic bacteria that are effective in degrading a variety of contaminants. Further study on the isolates from cluster 2 was carried out based on their phylogenetic analysis. Phylogenetic analysis of 28 bacterial isolates from cluster 2 based on the 1500bp sequences from 16S rDNA genes using MRBAYES confirmed all isolates as being hydrocarbonoclastic, providing supportive evidence that isolates from cluster 2 have a potential use in bioremediation. This approach could improve both the speed and efficiency of the commercial bioremediation process.     

Rhamnolipid production using Shewanella seohaensis BS18 and evaluation of its efficiency along with phytoremediation and bioaugmentation for bioremediation of hydrocarbon contaminated soils

Abstract

This study reports the combined use of a rhamnolipid type biosurfactant (BS) along with phytoremediation and bioaugmentation (BA) for bioremediation of hydrocarbon-contaminated soils. Bacterial isolates obtained from hydrocarbon contaminated soil were screened for rhamnolipid production and isolate BS18, identified as Shewanella seohaensis, was selected for bioremediation experiments. Growth of BS18 in mineral salt medium (MSM) with diesel oil as the carbon source showed a maximum biomass of 8.2?g L^?1, rhamnolipid production of 2.2?mg g^?1 cell dry weight, surface tension reduction of 28.6?mN/m and emulsification potential (EI₂₄%) of 65.6. Characterization of rhamnolipid based on Fourier transmittance infrared (FTIR) analysis confirmed the presence of OH, CH₂/CH₃, C=O, and COO stretching vibrations, respectively, which are distinctive features of rhamnolipid type BSs. In bioremediation experiments, the lowest hydrocarbon concentration of 2.1?mg g^?1 of soil for non-sterilized soil and 4.3?mg g^?1 of soil for sterilized soil was recorded in the combined application of rhamnolipid, phytoremediation, and BA. This treatment also yielded the highest hydrocarbon degrading bacterial population (6.4 Log Cfu g^?1 of soil), highest plant biomass (8.3?g dry weight plant^?1), and the highest hydrocarbon uptake (512.3?mg Kg^?1 of plant).     

Microbiological characterization of a fuel-oil contaminated site including numerical identification of heterotrophic water and soil bacteria

Seven soil samples and seven groundwater samples from a site contaminated with fuel-oil were investigated using several chemical and microbiological techniques. In soil samples, 500 to 7,500 mg/kg of total hydrocarbons were found. These samples contained no n-alkanes but iso- and branched chain alkanes. No polychlorinated biphenyls could be detected. Microbiological investigations included estimations of total cell counts, viable cell counts on different media, and numbers of methylotrophic, denitrifying, sulphate reducing, anaerobic (with the exception of methanogenic organisms), and hydrocarbon degrading bacteria. Viable and hydrocarbon degrading bacteria were found in all samples. A total of 1,366 pure cultures was characterized morphologically and physiologically and identified by numerical identification using a data base of more than 4,000 reference strains. Groundwater samples were dominated by gram-negative bacteria of the generaPseudomonas, Comamonas, Alcaligenes, andAcinetobacter, which were also found in soil samples. In addition, more grampositive bacteria belonging to the generaArthrobacter, Nocardia, andBacillus could be isolated from soil samples.     

采用酶液活性测定法快速筛选及构建石油烃高效降解菌系

快速筛选复杂有机物降解微生物混合菌系,在污染物治理过程中具有重要的实践意义.本研究首次尝试利用MicroResp^TM技术分析微生物酶液活性的方法,快速标定高效降解菌及混合菌系的石油烃降解能力,并采用传统的摇瓶培养检测法予以验证.通过微生物胞内、胞外及混合酶液的活性分析,考察了不同酶系(胞外、胞内及混合酶液)、菌系对石油烃分子的降解情况.结果表明: 结合MicroResp^TM技术的酶液活性测定法能够快速检测石油烃代谢酶系的降解能力,其灵敏度好、通量高,与传统的菌株摇瓶培养方法的检测结果基本一致.其中,7株菌株的120种全组合菌系活性测定试验在12 h周期内1次完成.筛选周期由传统摇瓶培养所需的7 d缩短10倍以上.以酶活性测定结果为指导设计的复配菌系具有较高的降解效率,最高石油烃降解率达(56.1±1.6)%.表明本筛选方法精度高、通量高,可用于石油烃降解功能菌系的构建.