混合菌系QZ-10对原油的作用性能表征及现场清防蜡应用

[背景]油藏环境中呈单相液态的原油,在开采、运送的过程中由于温度、压力及流动条件的变化,石蜡不断从原油中析出,造成井筒、管线的结蜡.微生物清防蜡作为一项新兴的技术,受到广泛的关注,但是需要根据现场的环境条件施用合适的微生物清防蜡菌液.[目的]利用来源于青海油田盐碱油藏环境的混合菌系QZ-10,有效解决油井结蜡的问题并探...     

Use of TLC‐FID and GC‐MS/FID to examine the effects of migratory state,diet and captivity on preen wax composition in White‐throated Sparrows Zonotrichia albicollis

Preen wax is important for plumage maintenance and other functions. Its chemical composition is complex, and separating and quantifying its components, commonly by gas chromatography (GC), can be challenging. We present a simple analytical system consisting of thin‐layer chromatography/flame ionization detection (TLC‐FID) using a solvent system of 100% toluene to analyse the complex compound classes present in preen wax. We used GC and TLC‐FID to investigate the effects of migratory status, diet and captivity on the preen wax composition of White‐throated Sparrows Zonotrichia albicollis, and to measure the quantity of preen wax on the head, primary and tail feathers. White‐throated Sparrows produced preen wax containing only monoesters regardless of migratory state. The monoesters contained several isomers consisting of homologous series of fatty alcohols (C10–C20) and fatty acids (C13–C19) esterified together in different combinations to form monoesters with total carbon numbers ranging from C23 to C38. Weighted average monoester carbon number was greater in captive birds than in wild birds and was greater in captives fed a formulated diet enriched with sesame oil than in birds fed the same diet enriched with fish oil. Captivity and migratory state also affected the complexity of the mixture of monoesters. There was significantly more preen wax on head feathers compared with primary and tail feathers. We suggest that among its many functions, preen wax may play a role in drag reduction by affecting the physical properties of feathers, and/or the fluid flow at their surfaces.     

花椒种籽油的含蜡量测定与脱蜡

花椒种籽油的含蜡量测定与脱蜡是长期困扰花椒种籽油处理的一项关键技术,本研究通过分析混合压榨制备的花椒种籽油,花椒籽种壳油及种仁油的含蜡量,研究了5种脱蜡方法脱除花椒籽油中蜡质的脱蜡效果,确定了脱除花椒籽油中蜡质的有效方法,研究结果表明,花椒籽油的含蜡量在15－20％,左右,而这些蜡质基本上都含在种壳油内一即种壳上,种仁油基本不含蜡质,脱除花椒籽油中蜡质的合理方法应是：（1）含蜡量相对较低的精制粗油可选用表面活性剂法脱蜡;（2）当含蜡量相对较高时,为降低脱蜡过程中油的耗损率可选用分步脱蜡法脱蜡;（3）需进行碱炼的油,可在碱炼过程中将蜡质与游离脂肪酸一半除去。     

Phenotypic Adaptations Help Rhodococcus erythropolis Cells during the Degradation of Paraffin Wax

During crude oil extraction, the reduction in temperature and pressure results in the precipitation of paraffin wax that contains 20–40 carbon chain hydrocarbons. The paraffin wax may accumulate inside production tubes, pipelines, and processing facilities, and also in tankers during petroleum transportation. There are few bacterial strains that are able to degrade solid substrates. In the present study, the biodegradation of paraffin is evaluated using Rhodococcus erythropolis cells. This bacterium is able to grow using paraffin wax from an oil refinery plant as the sole carbon source. The cells grow as a thick biofilm over the solid substrate, make scale‐like structures that increase the area of the initially smooth surface of paraffin, produce biosurfactants, and become more negatively charged than ethanol‐ or glucose‐grown cells. When paraffin wax is supplied as microparticles, to increase the cell–substrate contact area and to simulate paraffin precipitation, the cells also adjust the composition of the fatty acids of the phospholipids of the cellular membrane to decrease its fluidity and paraffin biodegradation increases considerably. The study suggests that the phenotypic adaptation of R. erythropolis cells may be used to degrade paraffin wax under real conditions.     

The origin of stomach oil in marine birds: Analyses of the stomach oil from six species of subantarctic procellariiform birds

The stomach oil produced by many marine birds of the order Procellariiformes is an important aspect of their breeding ecology. Fifty-seven samples of stomach oil from six species of subantarctic sea birds were examined by thin-layer and gas chromatography to determine the degree of variation in stomach oil composition between individuals of the same species. The wide variability detected, the typically marine composition of the component fatty acids and alcohols of the wax esters and triacyglycerols examined and the presence of pristane, squalene, and astaxanthin in the stomach oils all indicate that the bulk of the oil is derived directly from the food. This is in contrast to the nutritive fluids produced by secretion in several other groups of birds. Many of the stomach oils contain large amounts of wax ester and marine birds represent a significant link in the marine food web for the reconversion of zooplankton wax ester to triacylglycerol. No substantial offshore pollution by petroleum hydrocarbons was indicated by the samples; stomach oil samples from pelagic birds may be valuable in monitoring offshore pollution.     

Use of flax oil to influence honey bee nestmate recognition

Fatty acids, normally found in comb wax, have a strong influence on nestmate recognition in honey bees, Apis mellifera L. Previous work has shown that bees from different colonies, when treated with 16- or 18-carbon fatty acids, such as oleic, linoleic, or linolenic acids, are much less likely to fight than bees from two colonies when only one of the two is treated. Previous work also shows that the influence of comb wax on recognition has practical applications; transfer of empty comb between colonies, before merger of those colonies, reduces fighting among workers within the merged colony. Flax oil contains many of the same fatty acids as beeswax. Here, we tested the hypothesis that treatment of individual bees with flax oil affects nestmate recognition; the results proved to be consistent with this hypothesis and showed that treated bees from different colonies were less likely to fight than untreated bees. These results suggest that flax oil may be useful in facilitating colony mergers.     

Neutral Lipid Biosynthesis in Engineered Escherichia coli: Jojoba Oil-Like Wax Esters and Fatty Acid Butyl Esters

Wax esters are esters of long-chain fatty acids and long-chain fatty alcohols which are of considerable commercial importance and are produced on a scale of 3 million tons per year. The oil from the jojoba plant (Simmondsia chinensis) is the main biological source of wax esters. Although it has a multitude of potential applications, the use of jojoba oil is restricted, due to its high price. In this study, we describe the establishment of heterologous wax ester biosynthesis in a recombinant Escherichia coli strain by coexpression of a fatty alcohol-producing bifunctional acyl-coenzyme A reductase from the jojoba plant and a bacterial wax ester synthase from Acinetobacter baylyi strain ADP1, catalyzing the esterification of fatty alcohols and coenzyme A thioesters of fatty acids. In the presence of oleate, jojoba oil-like wax esters such as palmityl oleate, palmityl palmitoleate, and oleyl oleate were produced, amounting to up to ca. 1% of the cellular dry weight. In addition to wax esters, fatty acid butyl esters were unexpectedly observed in the presence of oleate. The latter could be attributed to solvent residues of 1-butanol present in the medium component, Bacto tryptone. Neutral lipids produced in recombinant E. coli were accumulated as intracytoplasmic inclusions, demonstrating that the formation and structural integrity of bacterial lipid bodies do not require specific structural proteins. This is the first report on substantial biosynthesis and accumulation of neutral lipids in E. coli, which might open new perspectives for the biotechnological production of cheap jojoba oil equivalents from inexpensive resources employing recombinant microorganisms.     

YdfH Identified as a Repressor of rspA by the Use of Reduced Genome Escherichia coli MGF-01

The effects of carnauba wax addition on the physical state of palm kernel oil-in-water emulsions were investigated. The oil-in-water emulsion (40 wt% oil + 60 wt% aqueous phase) kept the liquid state at 25°C irrespective of the presence or absence of carnauba wax in the oil phase. The emulsion containing the wax transformed from the liquid state to the solid state by shearing after storage for 20 h at 4°C, although the liquid-solid transition was not observed for the emulsion not containing the wax upon the same treatment. The viscoelasticity of the solid emulsions was demonstrated by small-deformation mechanical testing. Analysis of flow behavior of the emulsions showed that the change in physical properties of the emulsion containing the wax at 4°C was caused by the shearing at a low shear rate, around 50 s^?1–100 s^?1. According to the transition from the liquid state to the solid state of the emulsion containing the wax, the aggregation of oil droplets was found to occur to a large extent. The results of differential scanning calorimetry and surface pressure–surface area isotherms suggested that triglyceride molecules of palm kernel oil were more oriented at the oil–water interfaces in the emulsions after the wax addition. Based on these results, it is thought that carnauba wax is important in destabilization of palm kernel oil-in-water emulsions by modifying the physical state of the oil triglyceride molecules at the interfaces.     

Synthesis of oleyl oleate wax esters in Arabidopsis thaliana and Camelina sativa seed oil

Seed oil composed of wax esters with long‐chain monoenoic acyl moieties represents a high‐value commodity for industry. Such plant‐derived sperm oil‐like liquid wax esters are biodegradable and can have excellent properties for lubrication. In addition, wax ester oil may represent a superior substrate for biodiesel production. In this study, we demonstrate that the low‐input oil seed crop Camelina sativa can serve as a biotechnological platform for environmentally benign wax ester production. Two biosynthetic steps catalysed by a fatty alcohol‐forming acyl‐CoA reductase (FAR) and a wax ester synthase (WS) are sufficient to achieve wax ester accumulation from acyl‐CoA substrates. To produce plant‐derived sperm oil‐like liquid wax esters, the WS from Mus musculus (MmWS) or Simmondsia chinensis (ScWS) were expressed in combination with the FAR from Mus musculus (MmFAR1) or Marinobacter aquaeolei (MaFAR) in seeds of Arabidopsis thaliana and Camelina sativa. The three analysed enzyme combinations Oleo3:mCherry:MmFAR1?c/Oleo3:EYFP:MmWS, Oleo3:mCherry:MmFAR1?c/ScWS and MaFAR/ScWS showed differences in the wax ester molecular species profiles and overall biosynthetic performance. By expressing MaFAR/ScWS in Arabidopsis or Camelina up to 59% or 21% of the seed oil TAGs were replaced by wax esters, respectively. This combination also yielded wax ester molecular species with highest content of monounsaturated acyl moieties. Expression of the enzyme combinations in the Arabidopsis fae1 fad2 mutant background high in oleic acid resulted in wax ester accumulation enriched in oleyl oleate (18:1/18:1 > 60%), suggesting that similar values may be obtained with a Camelina high oleic acid line.     

Neutral lipid biosynthesis in engineered Escherichia coli: jojoba oil-like wax esters and fatty acid butyl esters

Wax esters are esters of long-chain fatty acids and long-chain fatty alcohols which are of considerable commercial importance and are produced on a scale of 3 million tons per year. The oil from the jojoba plant (Simmondsia chinensis) is the main biological source of wax esters. Although it has a multitude of potential applications, the use of jojoba oil is restricted, due to its high price. In this study, we describe the establishment of heterologous wax ester biosynthesis in a recombinant Escherichia coli strain by coexpression of a fatty alcohol-producing bifunctional acyl-coenzyme A reductase from the jojoba plant and a bacterial wax ester synthase from Acinetobacter baylyi strain ADP1, catalyzing the esterification of fatty alcohols and coenzyme A thioesters of fatty acids. In the presence of oleate, jojoba oil-like wax esters such as palmityl oleate, palmityl palmitoleate, and oleyl oleate were produced, amounting to up to ca. 1% of the cellular dry weight. In addition to wax esters, fatty acid butyl esters were unexpectedly observed in the presence of oleate. The latter could be attributed to solvent residues of 1-butanol present in the medium component, Bacto tryptone. Neutral lipids produced in recombinant E. coli were accumulated as intracytoplasmic inclusions, demonstrating that the formation and structural integrity of bacterial lipid bodies do not require specific structural proteins. This is the first report on substantial biosynthesis and accumulation of neutral lipids in E. coli, which might open new perspectives for the biotechnological production of cheap jojoba oil equivalents from inexpensive resources employing recombinant microorganisms.     

Wax production from renewable feedstock using biocatalysts instead of fossil feedstock and conventional methods

Background, aim, and scope

Using renewable feedstock and introducing biocatalysts in the chemical industry have been suggested as the key strategies to reduce the environmental impact of chemicals. The Swedish interdisciplinary research program “Speciality Chemicals from Renewable Resources—Greenchem” is aiming to develop these strategies. One target group of chemicals for Greenchem are wax esters which can be used in wood coatings to replace paraffin wax made from fossil crude oil. The aim of this study was to conduct a life cycle assessment of wax esters based on rapeseed oil produced by biocatalysts (enzymes). The scope was to compare the environmental performance of wax esters with paraffin wax produced by conventional methods.

Materials and methods

The study has a cradle-to-gate perspective and the functional unit is “1-kg wax product ready to use in a wood coating product.” Extensive data collection and calculations have been performed for the wax esters, whereas existing life cycle inventory data have been used for the paraffin wax.

Results

The energy input into the wax ester production is about one third of the energy input in paraffin wax production. However, the wax ester has a higher contribution to the global warming potential (GWP) due to high emissions of nitrous oxide from rapeseed cultivation. Referring to a cradle-to-grave perspective, including waste incineration, the contribution to the GWP will, however, be 3.5 times higher from paraffin wax. Wax ester makes a higher contribution to the acidification and eutrophication potential, due to emissions from soil from rapeseed cultivation, but five times lower contribution to the photochemical ozone creation potential. From a land-use perspective and a global warming point of view, it is more efficient to produce paraffin wax and grow high-yielding, short-rotation coppice (Salix) to replace fuel oil than it is to grow rapeseed for wax ester production.

Discussion

Overall, this study shows the importance of studying the environmental performance of a product not only from a gate-to-gate perspective but, instead, considering the environmental performance from cradle-to-gate. The biocatalytic production of the wax ester consumes less energy than the conventional chemical method, but the raw material step, cultivation of rapeseed contributes much to both acidification and eutrophication. When the waste treatment step is included, the contribution to GWP, however, for paraffin wax will be 3.5 times higher than for the wax ester.

Conclusions

From a gate-to-gate perspective, replacing conventional chemical processes by biocatalysts using enzymes leads to energy savings and reduces emissions. However, from a cradle-to-gate perspective, the use of renewable feedstock, such as rapeseed oil, may counteract some of these benefits. Concerning the GWP benefit from using renewable feedstock instead of fossil feedstock, the final waste treatment step must be included, thereby applying a cradle-to-grave perspective.

Recommendations and perspectives

The introduction of biocatalysts as a key strategy in reducing the environmental impact from the chemical industry is supported by the results in this study. On the other hand, it is not obvious that the key strategy of using renewable feedstock in chemical production per se leads to benefits concerning all environmental impact categories. Thus, much more attention needs to be paid to the choice of potential renewable feedstock options, the minimization of energy inputs, and the biological emissions from the soil in the cultivation of feedstock crops, improved gas cleaning in nitrogen fertilizer production plants, and the alternative use of the arable land, in optimizing the overall environmental benefits of an increased use of renewable feedstock in the chemical industry.     

Wax-incorporated Emulsion Gel Beads of Calcium Pectinate for Intragastric Floating Drug Delivery

The purpose of this study was to prepare wax-incorporated pectin-based emulsion gel beads using a modified emulsion-gelation method. The waxes in pectin–olive oil mixtures containing a model drug, metronidazole, were hot-melted, homogenized and then extruded into calcium chloride solution. The beads formed were separated, washed with distilled water and dried for 12 h. The influence of various types and amounts of wax on floating and drug release behavior of emulsion gel beads of calcium pectinate was investigated. The drug-loaded gel beads were found to float on simulated gastric fluid if the sufficient amount of oil was used. Incorporation of wax into the emulsion gel beads affected the drug release. Water-soluble wax (i.e. polyethylene glycol) increased the drug release while other water-insoluble waxes (i.e. glyceryl monostearate, stearyl alcohol, carnauba wax, spermaceti wax and white wax) significantly retarded the drug release. Different waxes had a slight effect on the drug release. However, the increased amount of incorporated wax in the formulations significantly sustained the drug release while the beads remained floating. The results suggest that wax-incorporated emulsion gel beads could be used as a carrier for intragastric floating drug delivery.     

Orange-derived stimuli regulating oviposition in the Mediterranean fruit fly

Abstract:  Some visual and olfactory host stimuli influencing oviposition of the Mediterranean fruit fly ( Ceratitis capitata Wiedemann) in orange fruit were investigated in a laboratory-reared strain of this species. Mated females of C. capitata were found to be attracted to the same extent by fragrant orange fruits and odourless sham oranges, while unmated females were notably less attracted than mated females by the above objects. Mated females laid significantly more eggs in orange-coloured than in whitish (optically neutral) spherical wax dummies (diameter, 9.0 cm) as well as in both orange-coloured and whitish wax dummies supplemented with high internal humidity (80–100%) compared with respective wax dummies supplied with low internal humidity (30–50%). The orange-like colour together with high internal humidity provides basic stimuli supporting adequate oviposition of C. capitata in oranges. The oviposition rate of female C. capitata was not significantly changed when graded dosages ranging from 0.004 to 19.6  μ l of orange peel oil per cm² were added to orange-coloured wax dummies, while oviposition was considerably subdued by addition of 3.9  μ l orange peel oil and completely disrupted by addition of 9.8 or 19.6  μ l of orange peel oil per cm² of whitish wax dummies. Oviposition of C. capitata in ripe orange fruit may thus be interpreted by the predilection of this tephritid species for an orange-coloured, glossy pericarp, being capable of counteracting the deterrent effect of the essential oil found in orange flavedo.     

Study on the role of Nocardia farcinica in enhancing the flow rate of crude oil

Microbial degradation of paraffin wax is an efficient method of removing wax deposition from pipelines and enhancing the flow rate of crude oil. The present study was carried out to isolate a potential paraffin-degrading organism from oil wells of Gujarat. Screening for bacteria-utilizing paraffin wax as the sole source of carbon was carried out using 2,6-dichlorophenol indophenol (DCPIP) dye as redox indicator. The selected organism was identified as Nocardia farcinica by 16S rRNA sequencing. Nocardia farcinica showed 100% degradation of heneicosane, 65.99% degradation of docosane, and 50.59% degradation of tricosane, the major components of paraffin wax, in 8 days, which was observed by gas chromatography. Eicosane (86%) and heneicosane (80%) were utilized more by the selected organism compared with octacosane (61%) and triacontane (58%) (DCPIP dye method). Gas chromatographic analysis revealed that the selected organism degraded 50% of paraffin crude oil in 10 days. To determine the ability of the selected organism to enhance flow rate, parameters such as viscosity (cps), surface tension (d/cm²), pour point (°C), and flow rate (min/2 ml) were determined, and the result showed significant reduction in all the parameters after the addition of Nocardia farcinica. The viscosity and surface tension of crude oil were reduced by 22 and 6.30 points, respectively, after the addition of Nocardia farcinica. Pour point and flow rate were reduced by 2 and 11 points, respectively, when compared with control. The above findings indicate that Nocardia farcinica isolated from crude oil plays a major role in enhancing the flow rate of crude oil.     

Molecular composition and surface properties of storage lipid particles in wax bean (Phaseolus vulgaris)

Lipid particles have been isolated from seeds of wax bean (Phaseolus vulgaris), a species in which starch and protein rather than lipid are the major seed storage reserves. These lipid particles resemble oil bodies present in oil-rich seeds in that > 90% of their lipid is triacylglycerol. Moreover, this triacylglycerol is rapidly metabolized during seed germination indicating that it is a storage reserve. The phospholipid surfaces of oil bodies are known to be completely coated with oleosin which prevents their coalescence, particularly during desiccation of the developing seed. This would appear to be necessary since lipid is the major storage reserve in oil seeds, and there are very few alternate types of storage particles in the cytoplasm of oil seed endosperm to provide a buffer against coalescence of oil bodies by isolating them from one another. The present study indicates that the surfaces of lipid particles from wax bean are not completely coated with oleosin and feature regions of naked phospholipid. This finding has been interpreted as reflecting the fact that lipid particles in wax been seeds are less prone to coalescence than oil bodies of oil-rich seeds. This arises because the individual lipid particles are interspersed in situ among highly abundant protein bodies and starch grains and hence less likely to come in contact with one another, even during desiccation of the developing seed.     

角蜡蚧和日本龟蜡蚧蜡泌物的超微结构及化学成分分析

采用扫描电镜和气相色谱/质谱联用技术研究了角蜡Ceroplastes ceriferus (Fabricius) 和日本龟蜡蚧C. japonicus Green蜡泌物的超微结构及化学成分。结果表明,这2种蜡蚧分泌蜡质和形成蜡壳的过程基本相似。在1、2龄期分泌的蜡质为“干蜡”,蜡壳为星芒状,虫体周缘的蜡芒均为2大节,与其2个龄期发育相对应,每一个大节又分为若干小节。同时,虫体背面中央蜡质堆积成帽状,也分为均匀的多层。由此说明泌蜡过程具有节律性。虫体周缘与蜡芒对应的突起区上分布着密集的刻点状腺孔,每一个腺孔分泌1根蜡丝,这在以往玻片标本中是观察不到的。雌性第3龄幼虫和成虫期,虫体分泌“湿蜡”,形成龟背状蜡壳,泌蜡腺孔主要为三格腺和四格腺。在肛突区发现了密集的泌蜡腺孔,排列为纵条纹状。从角蜡蚧蜡泌物甲酯化处理样品中检测到14个组分,从直接测试 (未经甲酯化处理) 样品中检测到14个组分;而从日本龟蜡蚧则分别检测到10个组分和25个组分。它们的主要成分是一系列高级的长链饱和与不饱和的烃、脂肪酸、脂肪醇、酯类、醛类以及杂环、多环或大环状化合物。对它们可能的生物生态学功能进行了讨论。     

白鱀豚额隆油的研究——Ⅰ.额隆油的脂肪酸组成

世界珍稀水兽白豚(Lipotes vexillifer.)是我国名贵特产之一,属齿鲸亚目(Odontoceti),淡水豚总科(Platanistoidea),白鱀豚科(Lipotidae)。白鱀豚生活在长江中、下游。借回声定位系统以探知外界情况(荆显英等,1981)。通常认为,在声发射过程中,齿鲸类饱含油分的额隆组织起着声透镜的作用。额隆的这种特殊生理机能,与其所含脂质的脂肪酸组成密切相关。1980年陆佩洪等报道了白鱀豚额隆油的酸价、碘价、皂化价、不皂化物及甘油三酯(比色法)的含量。有关白鱀豚额隆油的脂肪酸组成,迄今尚未研究。目前我们才开始进行该项工作。     

Utilization of gas oil by a yeast culture

A mixed yeast culture (Culture 4) was grown on representative gas oil samples as well as paraffin wax. Culture 4 was found to utilize n-paraffinic hydrocarbons almost quantitatively from most gas oil fractions; significant alteration of other hydrocarbon components was not detected. Generation times of 4.0–9.0hr. were typical during the exponential growth phase in fermentations with various gas oil fractions. Cell yields were 70–90% based on n-paraffin utilization. The culture appeared to exhibit maximum efficiency of n-alkane removal in the C₁₉ to C₂₄ range. The cells recovered from the fermentations contained 8.8–9.3% nitrogen. Paraffin wax also served as a suitable carbon source when dissolved in 2,6,10,14-tertramethylpentadecane (pristane). However, substrate utilization appeared to be incomplete.     

Crystallization and Gelation Behavior of Low- and High Melting Waxes in Rice Bran Oil: a Case-Study on Berry Wax and Sunflower Wax

Low-melting berry wax (BEW) has proven to be a good oil gelator with a positive contribution to the consistency and flexibility of the structured oil. Nevertheless, the properties of BEW and the corresponding oleogel have not yet been investigated in-depth. In this research, the difference in crystallization and gelling behavior between sunflower wax (SW), a high melting wax, and BEW, a low-melting wax, in rice bran oil (RBO) was investigated. The difference in melting and crystallization temperatures can be explained by the different chemical composition (long-chain wax esters in SW and short-chain fatty acids in BEW). The heterogeneity in crystal habits (unidirectional platelets versus microcrystalline particles) and polymorphism (orthorhombic versus hexagonal) are responsible for the varying gel strength and hardness of the respective SW- and BEW-oleogels. The microcrystalline BEW particles aligned and reorganized during 1-month storage at 5 °C, which leaded to an increase in the gel strength and hardness of BEW-oleogel. The gelling property of SW-oleogel however did not significantly differ after 4 weeks at 5 °C, despite of the appearance of spherulitic crystalline clusters. The changes in the physical properties of wax-based oleogels during storage time were further explored using differential scanning calorimetry, polarized light microscope, powder X-ray diffraction and rheology.