Variability in Pseudomonas aeruginosa Lipopolysaccharide Expression during Crude Oil Degradation

Bacterial utilization of crude oil components, such as the n-alkanes, requires complex cell surface adaptation to allow adherence to oil. To better understand microbial cell surface adaptation to growth on crude oil, the cell surface characteristics of two Pseudomonas aeruginosa strains, U1 and U3, both isolated from the same crude oil-degrading microbial community enriched on Bonny Light crude oil (BLC), were compared. Analysis of growth rates demonstrated an increased lag time for U1 cells compared to U3 cells. Amendment with EDTA inhibited U1 and U3 growth and degradation of the n-alkane component of BLC, suggesting a link between cell surface structure and crude oil degradation. U1 cells demonstrated a smooth-to-rough colony morphology transition when grown on BLC, while U3 cells exhibited rough colony morphology at the outset. Combining high-resolution atomic force microscopy of the cell surface and sodium dodecyl sulfate-polyacrylamide gel electrophoresis of extracted lipopolysaccharides (LPS), we demonstrate that isolates grown on BLC have reduced O-antigen expression compared with that of glucose-grown cells. The loss of O-antigen resulted in shorter LPS molecules, increased cell surface hydrophobicity, and increased n-alkane degradation.     

Variability in Pseudomonas aeruginosa lipopolysaccharide expression during crude oil degradation

Bacterial utilization of crude oil components, such as the n-alkanes, requires complex cell surface adaptation to allow adherence to oil. To better understand microbial cell surface adaptation to growth on crude oil, the cell surface characteristics of two Pseudomonas aeruginosa strains, U1 and U3, both isolated from the same crude oil-degrading microbial community enriched on Bonny Light crude oil (BLC), were compared. Analysis of growth rates demonstrated an increased lag time for U1 cells compared to U3 cells. Amendment with EDTA inhibited U1 and U3 growth and degradation of the n-alkane component of BLC, suggesting a link between cell surface structure and crude oil degradation. U1 cells demonstrated a smooth-to-rough colony morphology transition when grown on BLC, while U3 cells exhibited rough colony morphology at the outset. Combining high-resolution atomic force microscopy of the cell surface and sodium dodecyl sulfate-polyacrylamide gel electrophoresis of extracted lipopolysaccharides (LPS), we demonstrate that isolates grown on BLC have reduced O-antigen expression compared with that of glucose-grown cells. The loss of O-antigen resulted in shorter LPS molecules, increased cell surface hydrophobicity, and increased n-alkane degradation.     

Effects of sodium chloride on biodegradation of crude oils by two species ofAeromonas

Summary The biodegradation of five weathered crude oils by two species ofAeromonas, (B59-4 and E. BOB) was investigated in varying concentrations of sodium chloride. A minimal salts medium whose NaCl concentration increased serially by 0.5% w/v up to 1.5% w/v was used to investigate the growth of these strains in glucose, and their biodegradation of the crude oils. The latter was also investigated in fresh and aged sea water. Strain B59-4 was more potent than E. BOB in the degradation of all five crude oils and at all four levels of salt concentration tested. The amount of oil degraded by each strain increased initially to a maximum level at 0.5% w/v NaCl, but thereafter decreased with increasing salt concentration, and the patterns were similar to those of aged and fresh sea water, respectively. The Forties and Nigerian crude oils with lower specific gravity, were more readily degraded than the Libyan and Venezuelan with higher specific gravity. The growth of the two strains ofAeromonas in glucose and their biodegradation of crude oils was optimal at 0.5% w/v NaCl, and thereafter decreased with increasing salt concentration of the basal medium.     

Degradation of hydrocarbons and biosurfactant production by Pseudomonas sp. strain LP1

Pseudomonas sp. strain LP1, an organism isolated on the basis of its ability to grow on pyrene, was assayed for its degradative and biosurfactant production potentials when growing on crude, diesel and engine oils. The isolate exhibited specific growth rate and doubling time of 0.304 days⁻¹ and 2.28 days, respectively on crude oil (Escravos Light). The corresponding values on diesel were 0.233 days⁻¹ and 2.97 days, while on engine oil, were 0.122 days⁻¹ and 5.71 days. The organism did not show significant biosurfactant production towards crude oil and diesel, but readily produced biosurfactant on engine oil. The highest Emulsification index (E₂₄) value for the biosurfactant produced by LP1 on engine oil was 80.33 ± 1.20, on day 8 of incubation. Biosurfactant production was growth-associated. The surface-active compound which exhibited zero saline tolerance had its optimal activity at 50°C and pH 2.0.     

Effects of Temperature and Crude Oil Composition on Petroleum Biodegradation

The biodegradability of seven different crude oils was found to be highly dependent on their composition and on incubation temperature. At 20 C lighter oils had greater abiotic losses and were more susceptible to biodegradation than heavier oils. These light crude oils, however, possessed toxic volatile components which evaporated only slowly and inhibited microbial degradation of these oils at 10 C. No volatile toxic fraction was associated with the heavier oils tested. Rates of oil mineralization for the heavier oils were significantly lower at 20 C than for the lighter ones. Similar relative degradation rates were found with a mixed microbial community, using CO2 evolution as the measure, and with a Pseudomonas isolate from the Arctic, using O2 consumption as the measure. The paraffinic, aromatic, and asphaltic fractions were subject to biodegradation. Some preference was shown for paraffin degradation, especially at low temperatures. Branched paraffins, such as pristane, were degraded at both 10 and 20 C. At best, a 20% residue still remained after 42 days of incubation. Oil residues generally had a lower relative percentage of paraffins and higher percentage of asphaltics than fresh or weathered oil.     

Comparison of biodegradability of crude and fuel oils.

Crude and fuel oils were compared for ability to support growth of a mixed population of estuarine bacteria. A total of four oils, two crude and two fuel oils, were examined. It was found that each of the oils supported a unique population of bacteria and yeasts, with respect to generic composition. Low-sulfur, high-saturate, South Louisiana crude oil was found to be highly susceptible to degradation. In contrast, the dense, high-sulfur, high-aromatic, Bunker C fuel oil was strongly refractory to microbial degradation.     

Diversity and similarity of microbial communities in petroleum crude oils produced in Asia

To understand microbial communities in petroleum crude oils, we precipitated DNA using high concentrations of 2,2,4-trimethylpentane (isooctane) and purified. Samples of DNA from five crude oils, (Middle East, 3; China, 1; and Japan, 1) were characterized based upon their 16S rRNA gene sequences after PCR amplification and the construction of clone libraries. We detected 48 eubacterial species, one cyanobacterium, and one archaeon in total. The microbial constituents were diverse in the DNA samples. Most of the bacteria affiliated with the sequences of the three oils from the Middle East comprised similar mesophilic species. Acinetobacter, Propionibacterium, Sphingobium and a Bacillales were common. In contrast, the bacterial communities in Japanese and Chinese samples were unique. Thermophilic Petrotoga-like bacteria (11%) and several anaerobic-thermophilic Clostridia- and Synergistetes-like bacteria (20%) were detected in the Chinese sample. Different thermophiles (12%) and Clostridia (2%) were detected in the Japanese sample.     

杉木心材精油抑菌活性及其化学成分研究

通过水蒸气蒸馏法提取杉木心材精油,并进行柱层析分离、气-质联用分析和抑菌活性试验,比较分析了精油含量、化学组成和抑菌活性成分.结果表明,杉木心材精油含量为1.794～2.076(w/w);气-质联用分析共分离出47个色谱峰,鉴定出27个化合物(占精油总量的99%),其中主要成分为柏木脑(76.27%);杉木心材精油对大肠杆菌、金黄色葡萄球菌、枯草芽孢杆菌、伤寒沙门氏菌等均有较明显的抑制作用;柏木脑是杉木精油的主要抑菌活性成分.     

Microbial Community Composition in Crude Oils and Asphalts from the Kurdistan Region of Iraq

Abstract

To identify hydrocarbon-degrading microorganisms contributing to the formation of heavy oil we investigated the microbial community composition in different types of crude oils from oil-production facilities and in crude oil and asphalt from different natural seeps from the Kurdistan Region of Iraq (KRI). Crude oils from five out of six production facilities did not contain microorganisms detectable by 16S rRNA gene PCR amplicon sequencing likely reflecting a low microbial abundance in these samples. Crude oil and asphalt from the natural seeps hosted diverse microbial communities. The same phylotypes of uncultivated Deferribacteres and Thermodesulfobacteraceae were predominant community members across crude oils and asphalts from separate geographical locations. Soils surrounding seeps did not contain these phylotypes suggesting that they originate from the subsurface and although they seem commonly detected in hydrocarbon-rich environments their role in hydrocarbon-degradation is unknown. GC-MS analyses showed that mainly aromatic hydrocarbons were present in the crude oil and asphalt and that they were undergoing biodegradation - likely with sulfate and nitrate as terminal oxidants. In agreement, only bssA gene, but not assA gene-carrying microorganisms were detectable in the analyzed sampled. Overall our study identified several abundant uncultivated taxa with likely roles in transformation of nitrate, sulfate and hydrocarbons.     

The role of indigenous bacterial and fungal soil populations in the biodegradation of crude oil in a desert soil

The biodegradation capacity of indigenous microbial populations was examined in a desert soil contaminated with crude oil. To evaluate biodegradation, soil samples supplemented with 5, 10 or 20% (w/w) of crude oil were incubated for 90 days at 30 °C. The effect of augmentation of the soil with vermiculite (50% v/v) as a bulking agent providing increased surface/volume ratio and improved soil aeration was also tested. Maximal biodegradation (91%) was obtained in soil containing the highest concentration of crude oil (20%) and supplemented with vermiculite; only 74% of the oil was degraded in samples containing the same level of crude oil but lacking vermiculite. Gas chromatograms of distilled fractions of crude oil extracted from the soil before and after incubation demonstrated that most of the light and part of the intermediate weight fractions initially present in the oil extracts could not be detected after incubation. Monitoring of microbial population densities revealed an initial decline in bacterial viable counts after exposure to oil, presumably as a result of the crude oil’s toxicity. This decline was followed by a steep recovery in microbial population density, then by a moderate increase that persisted until the end of incubation. By contrast, the inhibitory effect of crude oil on the fungal population was minimal. Furthermore, the overall increased growth response of the fungal population, at all three levels of contamination, was about one order of magnitude higher than that of the bacterial population.     

Diet fat influences liver plasma-membrane lipid composition and glucagon-stimulated adenylate cyclase activity.

Rats were fed diets that differed in fatty acid composition or in the proportion of energy derived from fat to determine if alteration of dietary fat intake influences the structural lipid composition of liver plasma membrane and the expression of an associated hormone-receptor-mediated function. Weanling rats were fed 9% (w/w) or 20% (w/w) low-erucic acid rape-seed oil or 9% (w/w) soya-bean oil for 24 days. Plasma membranes were isolated and the effect of diet fat on the fatty acid composition of phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol and sphingomyelin was determined. Diet fat significantly altered total saturated and (omega-9) and (omega-6)-unsaturated fatty acid composition in addition to the (omega-6)- to (omega-3)-unsaturated fatty acid ratio in these polar lipids. Feeding the high-fat diet increased the (omega-6)- to (omega-3)-unsaturated fatty acid ratio and the (omega-9)-unsaturated fatty acid content in all lipids except sphingomyelin. Assay of glucagon-stimulated adenylate cyclase activity at both high and low glucagon concentrations indicated that high-fat intake also decreased cyclic AMP formation. In a second experiment the fat intake was held constant (40% of energy) and oleic acid was substituted for linoleic acid by blending high- and low-linoleic acid-type safflower oils. This experiment established that a dose-response relationship exists between dietary intake of fatty acid and the fatty acid composition of plasma-membrane phospholipids. Specific diet-induced transitions in membrane phospholipid fatty acid composition were paralleled by changes in glucagon-stimulated adenylate cyclase activity. This study suggests that transitions in dietary fat intake can alter a hormone-receptor-mediated enzyme function in vivo by changing the surrounding lipid environment.     

Preliminary studies on continuous recovery and purification of the penicillin acylase under pseudo-affinity conditions using phenyl-Sepharose gel

A continuous system for the recovery and purification of the penicillin acylase from crude extracts by recycling phenyl-Sepharose gel through three agitated vessels with disc filters of stainless steel is presented. The penicillin acylase present in the crude extract was absorbed into the phenyl-Sepharose gel under pseudo-affinity conditions (16% w/v of ammonium sulphate). After gel washing under the same conditions in the second vessel, enzyme desorption was performed using the same salt but at a lower concentration (6% w/v) in the third vessel. The preliminary studies reported here occurred without experimental difficulties, even at a gel concentration as high as 40% (v/v). The recovery of the penicillin acylase was achieved with high yield (74%), but a low purification factor of 2.4 was obtained owing to the use of a crude extract with low specific activity.     

Effects of amendments on biodegradation of crude petroleum by sediment bacteria from Bonny River Estuary

The biodegradation of Bonny light crude petroleum by bacteria in batch culture was enhanced by the addition to culture media, of 0.2 mg of urea and soya bean lecithin per 100 ml of crude oil, sediment and water mixture. Biodegradation was found to be purely an aerobic process. There was a direct relationship between hydrocarbon content and proportion (%) of total heterotrophic count that was capable of growing on crude petroleum as sole carbon and energy source.     

Development and optimization of fractional precipitation for the pre-purification of (+)-dihydromyricetin

A novel fractional precipitation process, both simple and efficient, was developed for producing (+)-dihydromyricetin in high purity and high yield from crude extracts. The optimal acetone composition in water, initial (+)-dihydromyricetin concentration in crude extract, storage temperature, storage time, and pH were 1/5 (v/v), 0.1 g/mL, 4°C, 32 h, and 9.0, respectively. Crude extracts were efficiently pre-purified using fractional precipitation of (+)-dihydromyricetin by differences of solubility in an acetone solution, increasing purity from 55.0 to over 84.9% with an overall yield of 97.5%. The use of fractional precipitation for pre-purification allowed for rapid and efficient separation of (+)-dihydromyricetin from interfering compounds and dramatically increased the purity of crude (+)-dihydromyricetin for subsequent purification steps.     

Fractional precipitation for paclitaxel pre-purification from plant cell cultures of Taxus chinensis

Fractional precipitation is a simple and efficient procedure for pre-purification of paclitaxel. The optimal methanol composition in water, paclitaxel content in crude extract, storage temperature, and time were 61.5% (v/v), 0.5% (w/v), 0°C, and 3 days, respectively. As purity of the paclitaxel in the crude extract increases, the purity and yield of paclitaxel from fractional precipitation increases. This pre-purification process serves to minimize the solvent usage, size and complexity of the HPLC operations for paclitaxel purification. This process is readily scalable to a pilot plant and eventually to a production environment where multikilogram quantities of material are expected to be produced.     

Characteristics and composition of Parkia biglobbossa and Jatropha curcas oils and cakes

Parkia biglobbossa (PKBS) and Jatropha curcas (JTC) seeds were analysed for their proximate composition. The seeds oils were analysed for fatty acid, lipid classes, sterols and physicochemical characteristics. Proximate analysis revealed that the percentage crude protein, crude fat and moisture in PKBS were 32.40%, 26.525% and 10.18% respectively and 24.60%, 47.25% and 5.54% in JTC. Campesterol, stigmasterol, beta-sitosterol, Delta5-avenasterol and Delta7-stigmasterol were identified in the seed oils, but beta-sitosterol was most abundant, constituting 71.9% in JTC and 39.5% in PKBS. JTC oil had 72.7% unsaturated fatty acids with oleic acid predominating, while PKBS had 62% unsaturated fatty acids with linoleic acid being the most abundant. Results of lipid classes showed triglyceride as the dominant lipid species in the seed oils. Physicochemical analysis of the seed oils showed that they could be classified as semi drying oils and that they could be found applicable in alkyd resin and soap manufacture.     

Current Prospects on Production of Microbial Lipid and Other Value-Added Products Using Crude Glycerol Obtained from Biodiesel Industries

Production of microbial lipids using crude glycerol from the biodiesel industry is reviewed in this paper. Approximately 10 wt.% of crude glycerol is obtained for every batch of biodiesel. The crude glycerol accumulated contains various impurities and hence cannot be used for any commercial applications without further purification. Its conversion via biological and chemical routes into valuable products has been studied by different researchers. Varieties of fungal, yeasts, and algal species have been used to produce microbial lipids from crude glycerol. However, research focus on screening a robust industrial oleaginous strain capable of doing this is still on-going. Due to its chemical similarity to vegetable oils, microbial lipids are considered a potential renewable feedstock for biodiesel production and for applications in food and pharmaceutical industries. Its conversion to polyols and subsequently to biobased polymers is also being explored. The rising price of vegetable oils, increasing energy demands, growing environmental concerns, and availability of crude glycerol as a cheap carbon substrate result in considerable potential for the application of these processes in the future.     

Degradation of mineral oils by a selected microbial association

A series of microbial associations capable of degrading various petroleum oils, emulsols, and crude oil were obtained by selection during periodic or continuous cultivation. The formation of these associations and oil-product degradation occurred most efficiently during aerobic flow cultivation. Under these conditions, oils were degraded by 92% on average. The microbial degradation of a petroleum oil depended on its brand, concentration, emulsification, and aeration.     

Filtration performance of microporous ceramic supports

The use of inorganic membranes in pollution treatment is actually limited by the cost of such membranes. Advantages of inorganic membranes are their chemical, thermal and pH properties. The purpose of this work was the development of microporous ceramic materials based on clay for liquid waste processing. The supports or ceramic filters having various compositions were prepared and thermally treated at 1100 °C. The results show that, at the temperature studied, porosity varied according to the support composition from 12% for the double-layered (ceramic) support to 47% for the activated carbon- filled support with a mean pore diameter between 0.8 and 1.3 μm, respectively. Volumes of 5 l of distilled water were filtered tangentially for 3 h under an applied pressure of 3.5 and 5.5 bar. The retention of tubular supports prepared was tested with molecules of varying size (Evans blue, NaCl and Sacharose). The study of the liquid filtration and flow through these supports showed that the retention rate depends on support composition and pore diameter, and solute molecular weight. The S1 support (mixture of barbotine and 1% (w/w) activated carbon) gave a flux for distilled water of 68 L/m² h while the double-layered support resulted in a flux of 8 L/m² h for the same solution at the pressure of 3.5 bar. At a pressure of 5.5 bar an increase in the distilled water flux through the various supports was observed. It was significant for the S1 support (230 L/m h).     

An overview of lipid metabolism in yeasts and its impact on biotechnological processes

High energy prices, depletion of crude oil supplies, and price imbalance created by the increasing demand of plant oils or animal fat for biodiesel and specific lipid derivatives such as lubricants, adhesives, and plastics have given rise to heated debates on land-use practices and to environmental concerns about oil production strategies. However, commercialization of microbial oils with similar composition and energy value to plant and animal oils could have many advantages, such as being non-competitive with food, having shorter process cycle and being independent of season and climate factors. This review focuses on the ongoing research on different oleaginous yeasts producing high added value lipids and on the prospects of such microbial oils to be used in different biotechnological processes and applications. It covers the basic biochemical mechanisms of lipid synthesis and accumulation in these organisms, along with the latest insights on the metabolic processes involved. The key elements of lipid accumulation, the mechanisms suspected to confer the oleaginous character of the cell, and the potential metabolic routes enhancing lipid production are also extensively discussed.