Effect of <Emphasis Type="Italic">Origanum vulgare</Emphasis> L. Essential Oil on Growth and Lipid Profile of <Emphasis Type="Italic">Yarrowia lipolytica</Emphasis> Cultivated on Glycerol-Based Media

Aim of the present study was to identify the chemical composition of Origanum vulgare L. essential oil and to investigate its effect on the biochemical behavior of the yeast Yarrowia lipolytica. Twenty-three compounds were identified by GC–MS analysis, representing the 92.3% (w/w) of the total essential oil of the plant. Carvacrol (56.3%) and thymol (16.4%) were the major components. Additionally, shake-flask cultivations of Y. lipolytica were performed, with various essential oil additions (0.05–2 mL/L of medium) on glycerol-based media. Growth was affected even at low concentrations (0.05 mL/L), while in higher essential oil concentrations, strong inhibition phenomena were observed. A tolerance-threshold concentration for the strain was hence established at 0.15 mL/L of oil. Furthermore, the presence of the essential oil in the culture medium resulted in changes in the composition of the intra-cellular lipids of the yeast. Specifically, oil addition to nitrogen-limited cultures to a level >0.15 mL/L caused a substantial increase in the percentage of saturated fatty acids (C16:0 and C18:0) in the lipid composition of the yeast Y. lipolytica.     

Process for production of arachidonic acid concentrate by a strain of mortierella alpina

Various Mortierella alpina fungi were screened for their capacities to produce arachidonic acid. A strain of M. alpina was found to show the highest productivity. Arachidonic acid content of biomass and overall yield per litre of culture was highest in soya flour supplemented medium which produced dispersed mycelium. When the glucose concentration in the medium was varied from 30 to 100 g/L, biomass, lipid, arachidonic acid content of biomass and arachidonic acid yield increased with increasing glucose concentration. Several natural oils, when added to the growth medium, stimulated arachidonic acid production. After fermentation in a 20-L fermenter under optimal culture conditions, the arachidonic acid yield was 5.3 g/L, representing 34.2% w/w of total fatty acids and 13.7% w/w of biomass. An extract containing 72.5% w/w arachidonic acid was prepared from the recovered mycelium.     

Production of 8,11,14,17-cis-eicosatetraenoic acid by Δ5 desaturase-defective mutants of an arachidonic acid-producing fungus, Mortierella alpina

Δ5 Desaturase-defective mutants of an arachidonic acid-producing fungus, Mortierella alpina 1S-4, accumulate large amounts of 8,11,14,17-cis-eicosatetraenoic acid (20:4ω3) when grown with linseed oil. One of the mutants, the S14 strain, produced 1.65 mg of 20:4ω3 per mL of culture medium (corresponding to 66.0 mg/g dry mycelia and 11.6% of total cellular fatty acids) when grown in a medium containing 1% glucose, 1% yeast extract, and 4% linseed oil methyl ester at 28°C for 2 d, and then at 16°C for 7 d. In a bench-scale fermentation in a 5-L jar fermenter, 20:4ω3 production reached 1.60 g/L of culture medium on the eighth day (corresponding to 77.3 mg/g dry mycelia and 26.0% of total cellular fatty acids). The cellular lipids of the S14 strain comprised 75.8% triacylglycerol (TG), 6.7% diacylglycerol, and 13.3% phospholipids (PL). The percentage of 20:4ω3 was higher in PL than in TG, and highest in phosphatidylcholine (32.6%).     

Antifungal Activities and Chemical Composition of Wood and Leaf Essential Oils from Cunninghamia konishii

Abstract

In this study antifungal activities of essential oils from wood and leaf and their constituents of Cunninghamia konishii against four wood decay fungi and six plant pathogenic fungi were investigated. GC and GC-MS analyses show that the major compounds of wood essential oil were cedrol and α-pinene, while those of leaf essential oil were α-pinene and p-cymene. Antifungal tests demonstrated the wood oil from C. konishii used against Trametes versicolor, Lenzites betulina, Laetiporus sulphureus, and Gloeophyllum trabeum and leaf oil from C. konishii used against L. sulphureus had strong antifungal activities. Moreover, wood oil used against Rhizoctonia solani, Fusarium solani, Pestalotiopsis funereal, and Ganoderma australe also had strong antifungal activities. Among the seven constituents of wood oil, cedrol displayed the best antifungal properties, indicating that it may be used as potential antifungal agents for the control of fungal diseases in plants.     

Lipid and γ-linolenic acid accumulation in strains of zygomycetes growing on glucose

Strains of Zygomycetes, belonging to the genera Zygorhynchus, Mortierella, Rhizopus, Mucor, and Cunninghamella, when cultivated on glucose produced significant quantities of γ-linolenic acid (GLA). After exhaustion of the nitrogen source from the culture medium, all strains accumulated cellular lipids in concentrations ranging from 10 to 28% (oil/dry mycelium). However, in some strains after the depletion of the carbon source (glucose) from the culture medium, a reconsumption of the accumulated oil and synthesis of fat-free cell material was observed. Accumulation of large amounts of oil in the mycelium resulted in the production of oil with low GLA content. Rhizopus stolonifer strain LGAM (9)1, and Cunninghamella sp. strain LGAM (9)2 produced more than 30 mg GLA/g of dry cellular mass. Cunninghamella sp. accumulated 28.1% oil/dry cellular mass, which contained 11.9% GLA. The production of GLA was 260 mg/L of culture medium.     

Effect of Growth Temperature on the Physico-Chemical Characteristics of Fungal Oil

The fungus Fusarium oxysporum f. sp. lini (Bolley) has been found to be a potential source for biotechnological production of oil. When grown as static surface culture in the best synthetic basal medium containing 15% glucose and 0.4% ammonium nitrate as sole carbon and nitrogen sources, respectively, at optimum pH 5.9 and temperature 28°±1°C, it gives 0.4% biomass of high protein (23.7%) and oil (18.9%) content, within a period of 14 days. The oil appears to be of economic importance since it contains appreciable proportions of nutritionally significant unsaturated fatty acids and resembles edible palm oil in composition.     

玳玳花精油成分的研究

采用水蒸汽蒸馏法对玳玳花进行提取制得精油,应用GC—FTIR和GC—MS对精油进行分析,经研究共分离确认组分28种,玳玳花精油主要成份是芳樟醇、乙酸芳樟酯、芋烯、β-蒎烯、罗勒烯、α-松油醇、香叶醇、乙酸香叶酯、橙花醇、乙酸橙花酯、橙花叔醇、金合欢醇等。并用手性色谱柱对芳樟醇进行手性分离,芳樟醇中（-）-芳樟醇（左旋）占90％以上,同时对玳玳进行植物学鉴定,玳玳的学名为Citrus aurantium L．’Daidai’。     

Microbial conversion of an oil containing α-linolenic acid to an oil containing eicosapentaenoic acid

Mycelia of arachidonic acid-producing fungi belonging to the genusMortierella were found to convert an oil containing α-linolenic acid to an oil containing 5,8,11,14,17-cis-eicosapentaenoic acid (EPA). This conversion was observed when they were grown in a medium containing the oil, glucose and yeast extract at 28 C. On the screening of various oils, linseed oil, in which α-linolenic acid amounts to about 60% of the total fatty acids, was found to be the most suitable for EPA production. Under the optimal culture conditions, a selected strain,Mortierella alpina 20-17, converted 5.1% of the α-linolenic acid in the added oil into EPA, the EPA production reaching 1.35 g/l of culture broth (41.5 mg/g dry mycelia). This value corresponded to 7.1% (by weight) of the total fatty acids in the extracted lipids. The lipid was also found to be rich in arachidonic acid (12.3%). Other major fatty acids in the lipid were palmitic acid (4.4%), stearic acid (3.2%), oleic acid (13.5%), linoleic acid (13.7%), α-linolenic acid (38.5%) and γ-linolenic acid (0.9%).     

Conversions of olive mill wastewater‐based media by Saccharomyces cerevisiae through sterile and non‐sterile bioprocesses

BACKGROUND: Olive mill wastewaters (OMWs) are an important residue and several methods have been proposed for their treatment. RESULTS: Remarkable decolorization (~63%) and phenol removal (~34% w/w) from OMW was achieved. In glucose‐based flask sterile cultures, enrichment with OMWs increased ethanol and biomass production compared with cultures without OMWs added. Flask sterile and un‐sterilized cultures demonstrated similar kinetic results. Batch‐bioreactor trials performed showed higher ethanol and lower biomass quantities compared with the respective shake‐flask experiments, while cultures used under un‐sterilized conditions revealed equivalent results to the sterile ones. In non‐sterile bioreactor cultures, OMWs addition enhanced biomass production in comparison with culture with no OMWs added, whereas ethanol biosynthesis was not affected. The maximum ethanol quantity achieved was 52 g L^?1 (conversion yield per sugar consumed of 0.46 g g^?1) in a batch bioreactor non‐sterilized trial with OMW–glucose enriched medium used as substrate, that presented initial reducing sugars concentration at ~115 g L^?1. Fatty acid analysis of cellular lipids demonstrated that in OMW‐based media, cellular lipids containing increased concentrations of oleic and linoleic acid were produced in comparison with cultures with no OMWs added. CONCLUSIONS: S. cerevisiae simultaneously produced bio‐ethanol and biomass and detoxified OMWs, under non‐sterile conditions. © 2012 Society of Chemical Industry     

Lipids of oleaginous yeasts. Part II: Technology and potential applications

The process of lipid accumulation in the oleaginous yeasts cultivated in various fermentation configurations when either sugars and related compounds or hydrophobic substances are used as substrates is presented and kinetic models describing both de novo and ex novo lipid accumulation are analyzed. Technological aspects related with single cell oil (SCO) produced by oleaginous yeasts are depicted. The influence of culture parameters upon lipid production process is presented. Lipid production has been studied in batch, fed‐batch, and continuous cultivation systems using yeasts belonging to the species Lipomyces starkeyi, Rhodosporidium toruloides, Apiotrichum curvatum, Candida curvata, Cryptococcus curvatus, Trichosporon fermentans, and Yarrowia lipolytica. The potentiality of yeasts to produce SCO as starting material of 2nd generation biodiesel is indicated and discussed. Of significant importance is also the utilization of yeast lipids as substitutes of high added value exotic fats (e.g., cocoa butter). Lipid produced by the various yeasts presents, in general, similar composition with that of common vegetable oils being composed of unsaturated fatty acids, whereas cocoa butter is principally composed of saturated fatty acids, consequently the various strategies that are followed in order to increase the cellular saturated fatty acid content of the yeast lipid are presented and comprehensively discussed.     

Lipid production by oleaginous Mucorales cultivated on renewable carbon sources

Mortierella isabellina and Cunninghamella echinulata were cultivated on glucose‐, pectin‐, starch‐ and lactose‐based media. Culture on glucose at two initial C/N ratios favored lipid synthesis in the media with increased C/N. Starch was an adequate substrate for both molds, but lipid (in g/g of biomass) was produced in lower quantities compared with the glucose trial. Pectin was inadequate for C. echinulata whereas growth of M. isabellina was satisfactory (8.4 g/L), followed by moderate lipid production. Growth of C. echinulata on lactose was negligible, while that of M. isabellina was notable (9.5 g/L) although lipid in biomass was only 0.36 g/g. Hydrolytic enzymes (α‐amylase, polygalacturonase and β‐galactosidase) activities of both strains seemed to be low enough to saturate their metabolic capabilities. This seemed a major cause for the lower amount of lipid accumulated during growth on complex media compared with that on glucose. Cellular fatty acids of M. isabellina were oleic, linoleic and palmitic acid, while γ‐linolenic acid (GLA) was produced in low quantities. In C. echinulata grown on glucose, lactose or starch, GLA concentration was notable at the beginning and end of culture. Growth on pectin at the first growth steps was accompanied by the production of saturated fatty acids, the amount of which decreased thereafter.     

Use of Essential Oils From Various Plants to Change the Fatty Acids Profiles of Lipids Obtained From Oleaginous Yeasts

We studied the ability of seven essential oils to alter the fatty acid composition of lipids produced by an oleaginous yeast Rhodosporidium toruloides. All of the essential oils, except thyme, significantly increased the stearic acid content of the lipids. The amount of essential oils in the media determined the fatty acid composition obtained. Subsequently, we studied the effect of the major monoterpenes present in these essential oils. When R. toruloides was grown on limonene, a major monoterpene in orange essential oil, the composition of lipid obtained was found to be quite similar to natural orange essential oil. This proved that limonene has a major role in the changes in fatty acid profiles of the lipids. The effect of orange essential oil on another oleaginous yeast, Cryptococcus curvatus, was also carried out. Although the effect of the essential oil on the fatty acid composition and biomass (cell mass) was similar for both these two yeasts, the reduction of the activity of some enzymes involved in the metabolic pathways was quite different. From these results, it can be concluded that the effect of essential oils differs with species and it is possible to produce lipids with alternate fatty acid profiles suitable for different applications and with good market value.     

松节油加成反应的研究进展

松节油是世界上最主要的植物精油,具有很高的应用价值。α-蒎烯和β-蒎烯是松节油最主要的两个组成成分,以这两个化合物为原料,经过多种化学反应途径,可以合成众多具有高附加值的衍生化合物。然而,目前对上文提及的化学反应途径的系统介绍仍然比较缺乏。本文对α-蒎烯和β-蒎烯经过加成反应合成衍生物的研究进行了综述;介绍了α-蒎烯和β-蒎烯与氢气、水、氯化氢等8类化合物进行加成反应生成衍生物的合成方法的研究,并简述了这些衍生物的应用研究。展望了未来松节油衍生物合成研究的方向,指出合成具有特殊性能的衍生物具有重要意义。     

Lipid body formation by <Emphasis Type="Italic">Thraustochytrium aureum</Emphasis> (ATCC 34304) in response to cell age

The heterotrophic marine protist, Thraustochytrium aureum produces substantial amounts of polyunsaturated fatty acids (PUFAs). In the present investigation, changes in the lipid and fatty acid profiles of T. aureum were studied according to the culture age. T. aureum was grown in artificial sea water medium for 10 days at 25 °C in shake culture condition. One to 10 day old cell samples were analyzed for cell biomass production, total lipid content, fatty acid profile and lipid body formation. In all the samples tested, total lipid production was found to be directly proportional to the dry cell weight of T. aureum. In the early phase of cell growth, cell biomass production, lipid content and glucose consumption were found to be higher. Thin layer chromatographic analysis (TLC) of lipids showed the presence of triacylglycerol (TAG; 169 mg/g, 90%), phospholipids (PL; 83 mg/g, 66%) and sterol (ST; 6 mg/g, 5%), which were recorded at maximum levels in the early growth phase of the cells. The composition of PUFAs and saturated fatty acids (SFAs) of the cell biomass and lipid class components (TAG and PL) was identified by gas chromatographic analysis (GC). In the early phase of cell growth, production of PUFAs in the total fatty acids was found to have attained maximum levels (61.3%) in which docosahexaenoic acid alone showed higher content of occurrence (99.0 mg/g in total lipid; 65.2 mg/g in TAG and 41.0 mg/g in PL). In the middle phase of cell growth, palmitic acid production was found to be higher (36.7 mg/g in total lipid; 31.3 mg/g in TAG and 12.6 mg/g in PL). Transmission electron microscopic studies of the cells showed the presence of a membrane around the lipid bodies in the early phase of cell growth. TAG and PL were actively involved in the formation of lipid bodies in the cells of T. aureum. Large-sized lipid bodies accumulated in 3 day old cells which were then fragmented into smaller bodies in the late growth phase.     

Utilization of extracellular lipids by HT29/219 cancer cells in culture

Uptake and incorporation of long-chain fatty acids were studied in a human colorectal cancer cell line (HT29/219) grown in culture medium supplemented with either fetal calf serum (FSC) or horse serum (HS). The cells were grown for 120 h with no change of medium; the two major cellular lipid classes, the phospholipids and the triacylglycerols, were analyzed at regular time-points. We observed significant changes in the concentration of most fatty acids throughout culture, and differences in their composition when different sera were used to supplement the medium. Minimal levels of free fatty acids were found in the cells, indicating a very small “free fatty acid pool”. A major difference between the cells grown in media supplemented with different sera was the changes observed in concentrations of cellular polyunsaturated fatty acids during growth. In cells grown with FCS (in which 20∶4n−6 is present), the levels of this acid in the phsopholipid and triacylglycerol fractions declined rapidly during cell growth, suggesting further metabolism. In cells grown in medium supplemented with HS, 18∶2n−6 was the major polyunsaturated acid present. There was clear evidence that this acid accumulated in the cellular triacylglycerol and phospholipid fractions. Furthermore, its concentration did not decline during growth in culture, suggesting minimal conversion to other polyunsaturated n−6 acids. Our results suggest that fatty acids from additional sources in the medium, for example triacylglycerols and phospholipids associated with the lipoproteins, are taken up by the cells. There is also indication of cellular fatty acid synthesis, particularly of monounsaturated and saturated acids during the culture period. HT29/219 cells were shown to take up and incorporate radioactivity when trace amounts of [1-¹⁴C]-labeled arachidonic, linoleic or oleic acids were added to the culture medium. Most (80%) of the label was detected in cellular phospholipids and triacylglycerols, although the specific activities of these various fatty acids were different in the two lipid fractions.     

Cationic polymerization of β-pinene with the AlCl3/SbCl3 binary catalyst: Comparison with α-pinene polymerization

The polymerization of β-pinene with the AlCl₃/SbCl₃ binary catalyst was investigated in toluene at −40°C and was compared with that of α-pinene. The polymerization of β-pinene with AlCl₃ alone was very rapid and retarded on addition of SbCl₃, in sharp contrast to that of α-pinene where added SbCl₃ remarkably accelerated it to give relatively high molecular weight oligomers. Attempted copolymerization of the two isomers with the binary catalyst, in turn, induced their parallel homopolymerizations, indicating that the copolymerization was difficult due to the large difference in reactivity. The homopolymerizations with AlCl₃/SbCl₃ were not seriously affected by a sterically hindered base, 2,6-di-tert-butyl-4-methylpyridine (DTBMP); the initiating species, therefore, would be different in nature from a proton. © 1996 John Wiley & Sons, Inc.     

Effect of lipids and surfactants on extracellular lipase production by Yarrowia lipolytica

The production of extracellular lipase in submerged cultures of Yarrowia lipolytica CECT 1240 has been investigated. Several compounds have been added to the culture medium, in order to assess their efficiency as inducers of lipase production. First, the effect of triglycerides (olive oil, sunflower oil, tributyrin) and fatty acids (oleic acid) has been studied. The highest activity level was obtained with sunflower oil (58 U cm^?3), followed by olive oil (49 U cm^?3). The cultures with tributyrin and oleic acid attained similar activities (33 U cm^?3). Then, several surfactants (Tween 80, Triton X‐100, gum arabic, polyethylene glycol 200) were added to the cultures with sunflower oil, in an attempt to increase the levels of extracellular lipase activity. The obtained activities were slightly lower than those achieved without surfactants. The assay of a wide range of surfactant concentrations in the case of PEG‐200 (with which the highest activity levels had been attained) did not improve the results. This strain secreted lipase concentrations two‐fold higher and showed significantly different behaviour towards the presence of surfactants in the culture medium, compared with other wild‐type Yarrowia lipolytica strains. Copyright © 2003 Society of Chemical Industry     

Effect of long and medium chain length lipids upon aqueous solubility of α-tocopherol

The efficiency by which α-tocopherol is solubilized in vitro into mixed bile salt micelles containing different lipids was studied. Alterations in solubility due to addition to the incubation media of triglyceride, free fatty acid, monoglyceride, and lecithin of either long or medium chain length were examined. Results are expressed as a partition ratio between a micellar and an oil phase. The triglyceride of both long and medium chain length fatty acids greatly decreased the solubility of α-tocopherol in bile salt solutions. When added singly, monoglyceride and lecithin of long chain length fatty acids increased the α-tocopherol solubilized four- to fivefold; fatty acids of either chain length and medium chain monoglyceride when added singly had no significant effect upon the tocopherol solubilized. An additive effect was observed when a combination of long chain monoglyceride and lecithin was added. Addition of fatty acid to this combination, however, significantly decreased the α-tocopherol solubility into the micellar phase. Although the solubility of α-tocopherol was increased by all combinations of medium chain length polar lipids, except the fatty acid-monoglyceride pair, the effect was three to seven times less than for the corresponding long chain mixture.     

Production of 8,11-cis-eicosadienoic acid by a Δ5 and Δ12 desaturase-defective mutant derived from the arachidonic acid-producing fungus Mortierella alpina 1S-4

The Δ5 and Δ12 desaturase (DS)-blocked mutants of an arachidonic acid-producing fungus, Mortierella alpina 1S-4, were obtained. These mutants accumulated 8,11-cis-eicosadienoic acid (20:2n-9). One of the mutants, M226-9, in which Δ5 and Δ12 DS are perfectly blocked, produced 1.68 mg of 20:2n-9 per mL of culture medium (101 mg/g dry mycelia) and no 5,8,11-cis-eicosatrienoic acid (20:3n-9) when grown in a medium containing 4% glucose and 1% yeast extract at 28°C for 2 d and then at 12°C for 12 d. The mycelial lipid comprised 77.4% triacylglycerol (TG) and 9.8% phosphatidylcholine (PC), among others. TG contained 69.0% of the total 20:2n-9, whose percentage in total TG fatty acids was 15.9%. The highest percentage (44.4%) of 20:2n-9 was found in PC. The addition of olive oil to the culture medium enhanced the production of 20:2n-9.     

Extraction and Characterization of Montmorency Sour Cherry (Prunus cerasus L.) Pit Oil

Montmorency sour cherry (Prunus cerasus L.) pit oil (CPO) was extracted and characterized by various methods including: GC, LC–MS, NMR, thermogravimetric analysis (TGA), differential scanning calorimetry (DSC) and X‐ray powder diffraction (XRD). The oil gave an acid value of 1.45 mg KOH/g, saponification value of 193 mg KOH/g and unsaponifiable matter content of 0.72 %. The oil contained oleic (O) and linoleic (l ) acids as the major components with small concentrations of α‐eleostearic acid (El, 9Z,11E,13E‐octadecatrienoic acid) and saturated fatty acid palmitic (P) acid. The CPO contained six major triacyglycerols (TAG), OOO (16.83 %), OLO (16.64 %), LLO (13.20 %), OLP (7.25 %), OOP (6.49 %) and LElL (6.16 %) plus a number of other minor TAG. The TAG containing at least one saturated fatty acid constitute 33 % of the total. The polymorphic behavior of CPO as studied by DSC and XRD confirmed the presence of α, β′ and β crystal forms. The oxidative induction time of CPO was 30.3 min at 130 °C and the thermal decomposition temperature was 352 °C.