Biodegradability of diesel oil

In batch culture diesel oil was degraded rapidly, with a maximum growth rate (for a consortium of microorganisms) of 0.55 h^-1. The corresponding yield Y _SX was 0.1 Cmol/Cmol. In a continuous stirred tank reactor the maximum dilution rate was about 0.25 h^-1, with a yield of 0.3 Cmol/Cmol. With a residence time of 1 day 82% of the influent oil was degraded. In the batch reactor, of the mixture of linear and branched alkanes the linear alkanes were degraded fastest and with the highest yield. Only after most of the linear alkanes had disappeared were the branched alkanes consumed. In a CSTR a large part of the branched alkanes was not degraded.     

海洋石油降解微生物及其降解机理

微生物修复作为一种新型环保的生物修复技术,已成为海洋石油污染生物修复的核心技术。对海洋石油降解微生物的种类即细菌、蓝藻、真菌以及藻类进行了总结,对微生物对石油烃的降解途径与降解机理进行了综述。微生物降解烷烃的过程包括末端氧化、烷基氢过氧化物以及环己烷降解3种形式。微生物对芳香烃的降解是通过芳香烃被氧化酶氧化导致苯环开环来实现的。微生物对多环芳烃的降解是在单加氧酶或双加氧酶的催化作用下被最终降解为二氧化碳和水而被分解。并对影响石油烃降解微生物的因素包括温度、营养物质等因素进行了分析。     

油藏微生物及其压力适应机制

目前我国油田开发主要处于高含水后期,微生物驱提高石油采收率技术(MEOR)以低成本、环境友好等独特的优势引起了石油工业界的重视。实际上,经过半个多世纪的发展,MEOR已经成为提高采收率的重要前沿技术。高压是油藏的主要环境特征,在影响油藏微生物生存与活性等方面具有重要作用。本文从油藏及其微生物的主要特征、微生物对高压环境的适应机制以及高压下微生物降解烃的代谢特征等方面进行了综述。介绍了对油藏微生物资源、群落结构、微生物在油水相中分布的认识,微生物乳化原油机制,以及微生物在油藏厌氧环境中协同代谢、受温度和压力影响的特点,并列举了MEOR的矿场应用。在高压适应机制上,微生物主要通过改变和调整细胞膜结构、增加胞内脂质组分和表达胞内特殊酶等作用来实现对压力的适应;在高压下烃降解微生物代谢速率低于常压,而且耐压菌和嗜压菌具有不同的烃降解效率。     

A Model for diffusion controlled bioavailability of crude oil components

Crude oil is a complex mixture of several different structural classes of compounds including alkanes, aromatics, heterocyclic polar compounds, and asphaltenes. The rate and extent of microbial degradation of crude oil depends on the interaction between the physical and biochemical properties of the biodegradable compounds and their interactions with the non-biodegradable fraction. In this study we have systematically altered the concentration of non-biodegradable material in the crude oil and analyzed its impact on transport of the biodegradable components of crude oil to the microorganisms. We have also developed a mathematical model that explains and accounts for the dependence of biodegradation of crude oil through a putative bioavailability parameter. Experimental results indicate that as the asphaltene concentration in oil increases, the maximum oxygen uptake in respirometers decreases. The mathematically fitted bioavailability parameter of degradable components of oil also decreases as the asphaltene concentration increases.     

Oil-oxidizing activity of bacteria isolated from south Sakhalin coastal waters

Ability of bacteria isolated from the southern coastal waters of the Sakhalin Island to degrade various hydrocarbons was studied. The population of marine microorganisms grown on oil was heterogeneous in terms of hydrocarbon degradation. The rate of bacterial degradation of oil hydrocarbons was shown to correlate with their growth rate on the model medium. The degradation rates were higher for aromatic hydrocarbons than for alkanes. Based on our data, the studied bacteria were conditionally assigned to three groups: active, intermediately active, and passive degraders. Ability to oxidize oil was previously not reported for members of the genus Cobetia.     

Microbial communities involved in anaerobic degradation of alkanes

Saturated hydrocarbons are quantitatively the most abundant fraction among all petroleum hydrocarbons. Significant advances have been made in the understanding of the anaerobic biodegradability of alkanes in terms of the microorganisms involved and the biochemical pathways over the past two decades. They can be used as carbon and energy sources by diverse physiological groups of microorganisms (isolates or consortia) grown under chlorate-reducing, nitrate-reducing, sufidogenic or methanogenic conditions. Two general biochemical mechanisms have been proposed for the initial activation of alkanes including addition of fumarate and carboxylation. However, glycyl radical enzymes dependent fumarate addition which yields alkyl-substituted succinate appear to be the most commonly shared mechanism for the anaerobic attack of alkanes under various redox conditions by phylogenetically diverse microorganisms. The genes encoding the candidate alkylsuccinate synthase have been recently described in alkane-degrading sulfate- and nitrate-reducers as well as in hydrocarbon-rich environments. Alternative mechanisms may also be available depending on the alkane-degrading microbial community and electron acceptors utilized.     

Biodegradation of Oil Tank Bottom Sludge using Microbial Consortia

We present a rationale for the selection of a microbial consortia specifically adapted to degrade toxic components of oil refinery tank bottom sludge (OTBS). Sources such as polluted soils, petrochemical waste, sludge from refinery-wastewater plants, and others were used to obtain a collection of eight microorganisms, which were individually tested and characterized to analyze their degradative capabilities on different hydrocarbon families. After initial experiments using mixtures of these strains, we developed a consortium consisting of four microorganisms (three bacteria and one yeast) selected in the basis of their cometabolic effects, emulsification properties, colonization of oil components, and degradative capabilities. Although the specific contribution each of the former parameters makes is not clearly understood, the activity of the four-member consortium had a strong impact not only on linear alkane degradation (100%), but also on the degradation of cycloalkanes (85%), branched alkanes (44%), and aromatic and sulphur–aromatic compounds (31–55%). The effectiveness of this consortium was significantly superior to that obtained by individual strains, commercial inocula or an undefined mixture of culturable and non-culturable microorganisms obtained from OTBS-polluted soil. However, results were similar when another consortium of four microorganisms, previously isolated in the same OTBS-polluted soil, was assayed.     

Biobased production of alkanes and alkenes through metabolic engineering of microorganisms

Advancement in metabolic engineering of microorganisms has enabled bio-based production of a range of chemicals, and such engineered microorganism can be used for sustainable production leading to reduced carbon dioxide emission there. One area that has attained much interest is microbial hydrocarbon biosynthesis, and in particular, alkanes and alkenes are important high-value chemicals as they can be utilized for a broad range of industrial purposes as well as ‘drop-in’ biofuels. Some microorganisms have the ability to biosynthesize alkanes and alkenes naturally, but their production level is extremely low. Therefore, there have been various attempts to recruit other microbial cell factories for production of alkanes and alkenes by applying metabolic engineering strategies. Here we review different pathways and involved enzymes for alkane and alkene production and discuss bottlenecks and possible solutions to accomplish industrial level production of these chemicals by microbial fermentation.     

Identification of crude-oil components and microorganisms that cause souring under anaerobic conditions

Oil souring has important implications with respect to energy resources. Understanding the physiology of the microorganisms that play a role and the biological mechanisms are both important for the maintenance of infrastructure and mitigation of corrosion processes. The objective of this study was to identify crude-oil components and microorganisms in oil-field water that contribute to crude-oil souring. To identify the crude-oil components and microorganisms that are responsible for anaerobic souring in oil reservoirs, biological conversion of crude-oil components under anaerobic conditions was investigated. Microorganisms in oil field water in Akita, Japan degraded alkanes and aromatics to volatile fatty acids (VFAs) under anaerobic conditions, and fermenting bacteria such as Fusibacter sp. were involved in VFA production. Aromatics such as toluene and ethylbenzene were degraded by sulfate-reducing bacteria (Desulfotignum sp.) via the fumarate-addition pathway and not only degradation of VFA but also degradation of aromatics by sulfate-reducing bacteria was the cause of souring. Naphthenic acid and 2,4-xylenol were not converted.     

Degradation of car engine base oil by Rhodococcus sp. NDKK48 and Gordonia sp. NDKY76A

Two microorganisms (NDKK48 and NDKY76A) that degrade long-chain cyclic alkanes (c-alkanes) were isolated from soil samples. Strains NDKK48 and NDKY76A were identified as Rhodococcus sp. and Gordonia sp., respectively. Both strains used not only normal alkane (n-alkane) but also c-alkane as a sole carbon and energy source, and the strains degraded more than 27% of car engine base oil (1% addition).     

油藏微生物群落研究的方法学

油藏微生物群落的解析和认知是开发和应用微生物采油技术的基础。利用各种提高油藏微生物可培养性的方法和非培养技术解析不同油藏微生物的群落结构、功能和多样性,对定向调控油藏微生物群落、开发和应用有效微生物驱油技术具有重要的指导意义。通过调查新近发展的提高微生物可培养性的方法和措施以及不依赖于培养的分子微生物生态学技术,总结了油藏微生物群落研究方法学的最新进展。提高微生物可培养性的方法和措施主要通过模拟微生物的生存环境,减少富营养的毒害作用、添加信号分子维持微生物细胞间的作用和提供新型电子供体和受体等手段采用稀释法、高通量培养法等方法得以实现;不依赖于培养的分子微生物生态学技术主要包括荧光原位杂交、末端限制性片断长度多态性分析、变性梯度凝胶电泳和构建克隆文库等技术。这些方法学的进展为更有效的获得各种油藏微生物资源、调控油藏微生物群落以提高石油采收率提供理论指导。     

中药郁金挥发油成分及挥发油中蜡质成分研究

对中药郁金挥发油成分进行ＧＣ－ＭＳ－ＤＳ分析,共分离得到１３７个气相色谱峰,鉴定出７３种成分,其中３６种为首次鉴定出。首次,作者对其挥发油中蜡质成分进行分离,经ＧＣ－ＭＳ－ＤＳ分析得其主要成分为长直链烷烃和长直链脂肪酸。     

Migration of N-Alkanes from Diesel Oil along the Bacteria-Ciliates Trophic Chain

The data on the dynamics of the content and qualitative composition of intracellular alkanes in Paramecium caudatum Ehrenberg, 1838 cells feeding on bacteria with different levels of exposure to diesel oil are presented. The possibility of migration of alkanes from diesel oil along the trophic chain from bacteria to ciliates was demonstrated.__________Translated from Izvestiya Akademii Nauk, Seriya Biologicheskaya, No. 3, 2005, pp. 375–379.Original Russian Text Copyright © 2005 by Karetnikova, Zhirkova.     

Petroleum biodegradation in marine environments

Petroleum-based products are the major source of energy for industry and daily life. Petroleum is also the raw material for many chemical products such as plastics, paints, and cosmetics. The transport of petroleum across the world is frequent, and the amounts of petroleum stocks in developed countries are enormous. Consequently, the potential for oil spills is significant, and research on the fate of petroleum in a marine environment is important to evaluate the environmental threat of oil spills, and to develop biotechnology to cope with them. Crude oil is constituted from thousands of components which are separated into saturates, aromatics, resins and asphaltenes. Upon discharge into the sea, crude oil is subjected to weathering, the process caused by the combined effects of physical, chemical and biological modification. Saturates, especially those of smaller molecular weight, are readily biodegraded in marine environments. Aromatics with one, two or three aromatic rings are also efficiently biodegraded; however, those with four or more aromatic ring are quite resistant to biodegradation. The asphaltene and resin fractions contain higher molecular weight compounds whose chemical structures have not yet been resolved. The biodegradability of these compounds is not yet known. It is known that the concentrations of available nitrogen and phosphorus in seawater limit the growth and activities of hydrocarbon-degrading microorganisms in a marine environment. In other words, the addition of nitrogen and phosphorus fertilizers to an oil-contaminated marine environment can stimulate the biodegradation of spilled oil. This notion was confirmed in the large-scale operation for bioremediation after the oil spill from the Exxon Valdez in Alaska. Many microorganisms capable of degrading petroleum components have been isolated. However, few of them seem to be important for petroleum biodegradation in natural environments. One group of bacteria belonging to the genus Alcanivorax does become predominant in an oil-contaminated marine environment, especially when nitrogen and phosphorus fertilizers are added to stimulate the growth of endogenous microorganisms.     

Evolution des populations bactériennes dans les sédiments des marais maritimes de l'Ile Grande pollués par I'Amoco Cadiz Evolution of Bacterial Populations in Salt Marsh Sediments of Ile Grande Polluted by Amoco Cadiz

Microbial investigations on heterotrophic bacteria (MPN counts, adenylates charge, growth rate and enzymatic activities) and hydrocarbon utilizing microorganisms (MPN counts) were carried out during three years in the salt marshes of Ile Grande. The aim was to follow their recovery after oil pollution. Oil degradation is related to bacterial activity (depending on site, depth in the soil, redox level and hydrocarbon concentrations). The results for the three years show a significant enhancement of the microflora during the first year (corresponding to removal of n. alkanes) followed by a decrease in bacterial activity which persisted for over two years. The activity of the heterotrophic bacterial population is deeply disturbed compared with unoiled sites, and these shifts remain detectable three years after oiling. This study does not modify our previous estimation regarding the recovery of the salt marshes.     

Determining the identity and roles of oil-metabolizing marine bacteria from the Thames estuary, UK

Crude oil is a complex mixture of different hydrocarbons. While diverse bacterial communities can degrade oil, the specific roles of individual members within such communities remain unclear. To identify the key bacterial taxa involved in aerobic degradation of specific hydrocarbons, microcosm experiments were established using seawater from Stanford le Hope, Thames estuary, UK, adjacent to a major oil refinery. In all microcosms, hydrocarbon degradation was significant within 10 weeks, ranging from > 99% of low-molecular-weight alkanes (C(10)-C(18)), 41-84% of high-molecular-weight alkanes (C(20)-C(32)) and pristane, and 32-88% of polycyclic aromatic hydrocarbons (PAHs). Analysis of 16S rRNA sequences from clone libraries and denaturing gradient gel electrophoresis (DGGE) indicated that, except when incubated with fluorene, PAH-degrading communities were dominated by Cycloclasticus. Moreover, PAH-degrading communities were distinct from those in microcosms containing alkanes. Degradation of the branched alkane, pristane, was carried out almost exclusively by Alcanivorax. Bacteria related to Thalassolituus oleivorans (99-100% identity) were the dominant known alkane degraders in n-alkane (C(12)-C(32)) microcosms, while Roseobacter-related bacteria were also consistently found in these microcosms. However, in contrast to previous studies, Thalassolituus, rather than Alcanivorax, was dominant in crude oil-enriched microcosms. The communities in n-decane microcosms differed from those in microcosms supplemented with less volatile alkanes, with a phylogenetically distinct species of Thalassolituus out-competing T. oleivorans. These data suggest that the diversity and importance of the genus Thalassolituus is greater than previously established. Overall, these experiments demonstrate how degradation of different petroleum hydrocarbons is partitioned between different bacterial taxa, which together as a community can remediate petroleum hydrocarbon-impacted estuarine environments.     

厌氧产表面活性剂微生物提高原油采收率的研究进展

微生物强化采油(microbial enhanced oil recovery,MEOR)是近年来在国内外发展迅速的一项提高原油采收率技术。微生物在油藏中高效生产表面活性剂等驱油物质是微生物采油技术成功实施的关键之一。然而,油藏的缺/厌氧环境严重影响好氧表面活性剂产生菌在油藏原位的生存与代谢活性;油藏注空气会增加开采成本,且注入空气的作用时效和范围难以确定。因此,开发厌氧产表面活性剂菌种资源并强化其驱油效率对于提高原油采收率具有重要意义。本文综述了国内外近年来利用厌氧产表面活性剂微生物提高原油采收率的研究进展,简述了微生物厌氧产表面活性剂的相关驱油机理、菌种资源开发现状以及油藏原位驱油应用进展,并对当前的研究提出了一些思考。     

Learning and discrimination of individual cuticular hydrocarbons by honeybees (Apis mellifera)

In social insect colonies, recognition of nestmates, kinship, caste and reproductive status is crucial both for individuals and for the colony. The recognition cues used are thought to be chemical, with the hydrocarbons found on the cuticle of insects often cited as being particularly important. However, in honeybees (Apis mellifera) the role of cuticular hydrocarbons in nestmate recognition is controversial. Here we use the proboscis extension response (PER) conditioning paradigm to determine how well honeybees learn long-chain linear alkanes and (Z)-alkenes present on the cuticle of worker bees, and also how well they can discriminate between them. We found large differences both in learning and discrimination abilities with the different cuticular hydrocarbons. Thus, the tested hydrocarbons could be classified into those which the bees learnt and discriminated well (mostly alkenes) and those which they did not (alkanes and some alkenes). These well-learnt alkenes may constitute important compounds used as cues in the social recognition processes.     

Microbial Degradation of High-Molecular-Weight Alkanes

Measurements of biological O(2) demand showed that normal alkanes containing up to 44 carbon atoms were metabolized by microorganisms.     

石油中长链烷烃微生物降解及分子机制研究进展

中长链烷烃是石油烃中的重要组成部分,由于其疏水性强、黏度大、化学活性低、难降解,是地下原油黏度大、石油采收率低、泄漏后长期污染生态环境的重要原因,因此成为提高石油采收率和石油污染环境治理中的重要降解目标。微生物降解中长链烷烃作为一种新型高效的绿色技术日益受到重视。本文总结了微生物降解中长链烷烃的间期适应与转运过程,与转运过程相关的膜蛋白,微生物好氧与厌氧降解的代谢途径,以及好氧降解过程中的基因调控机制,并对微生物降解中长链烷烃的研究方向提出了展望,以期为后续的相关研究工作提供参考。