Phospholipids of two strains of dermatophyte,Arthroderma uncinatum

The phospholipid composition of a mutant strain of the dermatophyteArthroderma uncinatum was compared with that of the wild type. Phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, diphosphatidylglycerol, phosphatidylinositol, and phosphatidic acid were present, and there was marked variation in the amounts of phosphatidylcholine, phosphatidylserine, and phosphatidic acid in the two strains. Thus, the ratio of phosphatidylcholine (wild type) to phosphatidylcholine (mutant) was 2∶1; the ratio of phosphatidylserine (wild type) to phosphatidylserine (mutant) was 3∶1 and the ratio of phosphatidic acid (wild type) to phosphatidic acid (mutant) was 1∶5. In both strains, the predominant fatty acid was 18∶2, with 54.0% in the wild type and 46.7% in the mutant. Qualitatively, the same fatty acids, with the exception of C_20∶0, were found in all of the phospholipid classes; C_20∶0, both in the mutant strain (5.79%) and the wild type (trace amounts), was associated with phosphatidylserine. There was a significant difference in the rates of growth of the two strains; the mutant strain grew more rapidly than, and produced three times as much mycelium as, the wild type. The mutant strain also produced larger proportions of total lipid and phospholipid than the wild type; there was 20.5% total lipid and ca. 5% phospholipid in the mutant compared with 15.5% total lipid and 2.6% phospholipid in the wild type.     

Changes in Oil Content of Transgenic Soybeans Expressing the Yeast <Emphasis Type="Italic">SLC1</Emphasis> Gene

The wild type (Wt) and mutant form of yeast (sphingolipid compensation) genes, SLC1 and SLC1-1, have been shown to have lysophosphatidic acid acyltransferase (LPAT) activities (Nageic et al. in J Biol Chem 269:22156–22163, 1993). Expression of these LPAT genes was reported to increase oil content in transgenic Arabidopsis and Brassica napus. It is of interest to determine if the TAG content increase would also be seen in soybeans. Therefore, the wild type SLC1 was expressed in soybean somatic embryos under the control of seed specific phaseolin promoter. Some transgenic somatic embryos and in both T2 and T3 transgenic seeds showed higher oil contents. Compared to controls, the average increase in triglyceride values went up by 1.5% in transgenic somatic embryos. A maximum of 3.2% increase in seed oil content was observed in a T3 line. Expression of the yeast Wt LPAT gene did not alter the fatty acid composition of the seed oil.     

Phospholipids inDrosophila heads: Effects of visual mutants and phototransduction manipulations

A procedure was developed to label phospholipids inDrosophila heads by feeding radioactive phosphate (³²P_i). High-performance thin-layer chromatography showed label incorporation into various phospholipids. After 24 h of feeding, major phospholipids labeled were phosphatidylethanolamine (PE), 47%; phosphatidylcholine (PC), 24%; and phosphatidylinositol (PI), 12%.Drosophila heads have virtually no sphingomyelin as compared with mammalian tissues. Notable label was in ethanolamine plasmalogen, lysophosphatidylethanolamine, lysophosphatidylcholine and lysophosphatidylinositol. Less than 1% of the total label was in phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate. Other lipids labeled included phosphatidylserine, phosphatidic acid and some unidentified lipids. A time course (3–36 h) study revealed a gradual decrease in proportion of labeled PI, an increase in proportion of labeled PC and no obvious change in labeled PE. There were no significant differences in phospholipid labeling comparing theno receptorpotential (norpA) visual mutant and wild type under lightvs. dark conditions. However, overall³²P labeling was higher in the wild type fed in the light as compared to the dark and tonorpA either in light or dark. This suggests that functional vision facilitates incorporation of label. Differences in phospholipid labeling were observed between young and aged flies, particularly in lysophospholipids and poly-PI, implicating phospholipase A₂ function in recycling. Manipulations such as theouterrhabdomeresabsent andeyesabsent mutants and carotenoid deprivation failed to yield notable differences in phospholipid labeling pattern, suggesting that phospholipids important to vision may constitute only a minor portion of the total labeled pool in the head.     

Effects of oxidative stress on glycerolipid acyl turnover in rat hepatocytes

Lipid peroxidation was induced in freshly isolated rat hepatocytes by incubation in the presence of Fe³⁺, resulting in accumulation of thiobarbituric acid reactive substances. Analysis of lipid classes revealed that the levels and fatty acid compositions of the two major phospholipids, phosphatidylcholine (PC) and phosphatidylethanolamine (PE), remained unchanged but the levels of triacylglycerols (TAG) were significantly reduced, and some of their polyunsaturated fatty acids were selectively lost as the result of oxidant treatment. Acyl turnover in PC and PE as determined by ¹⁸O incorporation from H₂ ¹⁸O-containing media remained largely unchanged during oxidant treatment, while some increased turnover of the saturated fatty acids in TAG was observed. We hypothesize that constitutive recycling of membrane phospholipids rather than selective in situ repair eliminates peroxidized species of PC and PE, TAG could serve as an expendable fatty acid reserve, providing a limited but very dynamic pool of polyunsaturated fatty acids for the resynthesis of phospholipids.     

Characterization of the glycerolipid composition of a high‐palmitoleic acid sunflower mutant

The glycerolipid composition of a high‐palmitoleic acid sunflower (Helianthus annuus L.) mutant accumulating up to 20% of n‐7 fatty acids was studied. This line produces oil with a complex triacylglycerol (TAG) composition, containing species that have not been previously identified in sunflower. In this regard, palmitoleic acid was esterified in an unexpected way in the three positions of the TAG molecules. The polar glycerolipid composition of the mutant was also studied, in order to identify and quantify the changes in membrane lipids imposed by the sunflower enzymatic machinery during the accumulation of the unusual n‐7 fatty acids. The high‐palmitoleic mutant accumulated important quantities of n‐7 fatty acids in the polar lipid fraction, especially in the phosphatidylcholine lipid class. However, the total polar lipid content of these lines was not affected. On the other hand, the mutations responsible for the n‐7 lipid accumulation induced an important decrease in the oil yield of the new mutant.     

Lipid class composition of the protozoan Perkinsus marinus, an oyster parasite, and its metabolism of a fluorescent phosphatidylcholine analog

Perkinsus marinus is one of two important protozoan parasites of the eastern oyster, Crassostrea virginica. The other is Haplosporidium nelsoni. Lipids extracted from 7-d-old in vitro cultured P. marinus meronts, incubated with fluorescent-labeled phosphatidylcholine (FL PC) and nonincubated P. marinus meronts, were analyzed by a high-performance liquid chromatography (HPLC) system equipped with a diol phase column, in combination with thin-layer chromatography coupled with a flameionization detector (TLC/FID), and high-performance thin-layer chromatography (HPTLC). Various polar and neutral lipid classes were separated by HPLC using a two-gradient solvent system. Five polar lipid classes—phosphatidylcholine (PC), phosphatidylethanolamine (PE), cardiolipin (CL), sphingomyelin (SM), and phosphatidylserine (PS)—were identified from P. marinus extracts. Four neutral lipid classes—triacylglycerol (TAG), steryl ester (SE), cholesterol (CHO), and fatty alcohol—were distinguished. TLC/FID analysis of meront lipids showed that the weight percentages of PC, PE, CL, SM, PS/PI, TAG, SE, and CHO were 21, 10.7, 4, 2.3, 4.3, 48.7, 7.8, and 1.2%, respectively. HPLC and HPTLC analyses revealed the presence of two SM and PS isomers in P. marinus extracts. Perkinsus marinus effectively incorporated FL PC acquired from the medium and metabolized it to various components (i.e., free fatty acid, monoacylglycerol, diacylglycerol, TAG, PE, and CL). Uptake and interconversion of FL PC in P. marinus meronts increased with time. After 48 h the total uptake of fluorescence (FL) was 28.9% of the FL PC added to the medium, and 43% of the incorporated FL resided in TAG.     

Variation in fatty acid composition of the different acyl lipids in seed oils from fourSesamum species

Seeds from different collections of cultivatedSesamum indicum Linn. and three related wild species [specifically,S. alatum Thonn.,S. radiatum Schum and Thonn. andS. angustifolium (Oliv.) Engl.] were studied for their oil content and fatty acid composition of the total lipids. The wild seeds contained less oil (ca. 30%) than the cultivated seeds (ca. 50%). Lipids from all four species were comparable in their total fatty acid composition, with palmitic (8.2–12.7%), stearic (5.6–9.1%), oleic (33.4–46.9%) and linoleic acid (33.2–48.4%) as the major acids. The total lipids from selected samples were fractionated by thin-layer chromatography into five fractions: triacylglycerols (TAG; 80.3–88.9%), diacylglycerols (DAG; 6.5–10.4%), free fatty acids (FFA; 1.2–5.1%), polar lipids (PL; 2.3–3.5%) and steryl esters (SE; 0.3–0.6%). Compared to the TAG, the four other fractions (viz, DAG, FFA, PL and SE) were generally characterized by higher percentages of saturated acids, notably palmitic and stearic acids, and lower percentages of linoleic and oleic acids in all species. Slightly higher percentages of long-chain fatty acids (20∶0, 20∶1, 22∶0 and 24∶0) were observed for lipid classes other than TAG in all four species. Based on the fatty acid composition of the total lipids and of the different acyl lipid classes, it seems thatS. radiatum andS. angustifolium are more related to each other than they are to the other two species.     

Generation and remodeling of highly polyunsaturated molecular species of rat hepatocyte phospholipids

Freshly isolated rat hepatocytes were incubated for 20 min with [U-¹⁴C]glycerol in the presence or absence of unlabeled linoleic (18∶2n-6), arachidonic (20∶4n-6), or docosahexaenoic (22∶6n-3) acid, added as albumin complex in 10% ethanol. Most of the radioactivity (≈95%) recovered in hepatocyte lipids was present in phosphatidylcholine (PC), phosphatidylethanolamine (PF), and triacylglycerol (TAG). The presence of exogenous fatty acids resulted in (i) higher incorporation of [U-¹⁴C]glycerol, (ii) higher percentage of label in TAG, and (iii) enhanced formation of PC and PE molecular species bearing the exogenous fatty acid at both the sn-1 and sn-2 positions of glycerol. In each case, these molecular species contained 60 to 70% of the label in that lipid class. Further incubation of the cells for 40 and 80 min in the absence of labeled substrate and exogenous fatty acids resulted in a redistribution of label among PC and PE molecular species due to deacylation-reacylation at the sn-1 position of glycerol.     

Thermoxidative stability of triacylglycerols from mutant sunflower seeds

Thermoxidative stability was evaluated in triaclyglycerols (TAG) from the oils of the mutant sunflower lines CAS-3, CAS-4, and CAS-8 (with a high percentage of stearic acid), CAS-5 (with a high percentage of palmitic acid), all from standard highlinoleic genetic backgrounds, and the mutant sunflower line CAS-12 (with a high percentage of palmitic acid), from a high-oleic genetic background. These oils contained unusually high contents of TAG molecular species with one or two saturated fatty acids at the sn-1,3 positions. Purified total TAG devoid of tocopherols were subjected to controlled thermoxidative treatment at 180°C. Polymerized TAG were determined at 2-h intervals for 10 h. After this time, total polar compounds, oxidized TAG monomers, TAG dimers, and TAG oligomers were determined. TAG from highly saturated sunflower oils with levels of linoleic acid similar to those found in conventional sunflower oils (40–50%) showed enhanced thermal stability. In these TAG, the amount of polar compounds formed during the thermoxidative treatment was similar to that formed in the high oleic acid line. Excellent results were obtained for the TAG of the CAS-12 oil, which had the highest thermal stability, producing half the amount of polar compounds as the conventional line and less than two-thirds that of the high-oleic line.     

Effects of dietary triacylglycerol structure on triacylglycerols of resultant chylomicrons from fish oil- and seal oil-fed rats

We investigated the influence of the intramolecular fatty acid distribution of dietary triacyl-sn-glycerols (TAG) rich in n-3 polyunsaturated fatty acids (PUFA) on the structure of chylomicron TAG. Fish oil and seal oil, comparable in fatty acid compositions but with different contents of major n-3 PUFA esterified at thesn-2 position (20:5n-3, 46.6%, and 5.3%; 22:6n-3, 75.5%, and 3.8%, respectively), were fed to rats. Mesenteric lymph was collected and the chylomicrons were isolated by ultracentrifugation. The fatty acid composition of chylomicrons largely reflected the fatty acid composition of the oils administered. The intramolecular fatty acid distributions of the TAG fed were reflected in the chylomicron TAG as the fraction of the total contents observed in thesn-2 position of 20:5n-3 were 23.6 and 13.3%, and of 22:6n-3 were 30.6 and 5.4% for resultant chylomicrons following fish oil and seal oil administration, respectively. Thus, after seal oil administration, significant higher load of n-3 PUFA was esterified in thesn-1,3 positions of chylomicron TAG compared with fish oil administration (P<0.05).     

Triacylglycerol structure of human colostrum and mature milk

Because triacylglycerol (TAG) structure influences the metabolic fate of its component fatty acids, we have examined human colostrum and mature milk TAG with particular attention to the location of the very long chain polyunsaturated fatty acid on the glycerol backbone. The analysis was based on the formation of various diacylglycerol species from human milk TAG upon chemical (Grignard degradation) or enzymatic degradation. The structure of the TAG was subsequently deduced from data obtained by gas chromatographic analysis of the fatty acid methyl esters in the diacylglycerol subfractions. The highly specific TAG structure observed was identical in mature milk and colostrum. The three major fatty acids (oleic, palmitic and linoleic acids) each showed a specific preference for a particular position within milk TAG: oleic acid for thesn-1 position, palmitic acid for thesn-2 position and linoleic acid for thesn-3 position. Linoleic and α-linolenic acids exhibited the same pattern of distribution and they were both found primarily in thesn-3 (50%) andsn-1 (30%) positions. Their longer chain analogs, arachidonic and docosahexaenoic acids, were located in thesn-2 andsn-3 positions. These results show that polyunsaturated fatty acids are distributed within the TAG molecule of human milk in a highly specific fashion, and that in the first month of lactation the maturation of the mammary gland does not affect the milk TAG structure.     

Transfer of lipids between hemolymph and hepatopancreas in the shrimp Macrobrachium borellii

Crustancean lipids are transported in the hemolymph by an HDL. The hepatopancreas is the most important and active organ regarding lipid metabolism, so we studied the interchange of FA and acylglycerols between both components of the hepatopancreas-hemolymph system in the decapod crustacean Macrobrachium borellii. The hepatopancreas and a sole plasma lipoprotein were labeled by in vivo incubations with ¹⁴C palmitic acid injected into the hemolymph. Then they were incubated in vitro with unlabeled hepatopancreas and hemolymph, and the transfer of lipids between them was measured by radiochromatographic techniques. It was determined in vivo that more than 80% of the circulating palmitic acid was taken up by the hepatopancreas and incorporated into PC and TAG. Both classes of lipids, but mainly PC, were transferred back from tissues to the hemolymph. Lipid transfer was also demonstrated in vitro. The transfer of PC (30% of labeling) as well as that of FFA (48% of labeling) from hemolymph to hepatopancreas was determined. On the other hand, FFA were released more efficiently than the acylglycerols from intact hepatopancreas to hemolymph, and they were the only lipid transferred when the hepatopancreas had been previously washed.     

Lipidic characterization of full-grown amphibian oocytes and their plasma membrane-enriched fractions

The content and composition of neutral lipids and phosphoglycerides from full-grown prophase-arrestedBufo arenarum Hensel oocytes and from their ghost preparations were studied. The ghosts obtained are highly enriched in plasma membrane as suggested by the activity of 5′-nucleotidase, a marker enzyme, and the level of typical membrane components such as sphingomyelin, phosphatidylserine (PS), phosphatidylinositol (PI), and phosphatidic acid. In whole oocytes, triacylglyceride (TAG) comprises about 60% of the total lipids followed by phosphatidylcholine (PC), cholesterol, and phosphatidylethanolamine (PE). TAG and diacylglycerides have a similar unsaturation index. PC and PE account for about 80% of the phosphoglycerides in the whole oocyte and in their plasma membrane-enriched fractions. Arachidonic acid (20∶4n−6), 18∶0, and 16∶0 make up about 80 mol% of the total fatty acids in Pl in whole oocytes and ghost fractions. The unsaturation index in PS is higher in intact oocytes than in ghost preparations, probably owing to the significant amount of 20∶4n−6 which comprises 23 mol% of the total fatty acids in whole oocytes. The fatty acid profile in phosphatidic acid from whole oocytes is rather different from that in ghosts. Sphingomyelin contains mainly saturated and monounsaturated fatty acids, 24∶1 being the principal very long chain unsaturated fatty acid in both oocytes and ghosts.     

Comprehensive Identification of Principal Lipid Classes and Tocochromanols in Silkworm (Antheraea pernyi and Bombyx mori) Pupae Oils

A comprehensive identification of lipid compositions and tocochromanols in Antheraea pernyi (A. pernyi) and Bombyx mori (B. mori) pupae oil is reported in the present study. Fatty acid profiling shows that both oils contain high levels (79.67% vs 71.11%) of unsaturated fatty acids, especially linoleic acid and linolenic acid. Moreover, linolenic acid is preferentially enriched at the sn‐2 positions of triacylglycerols (TAGs). Liquid chromatograph‐mass spectrometry (LC‐MS) analysis demonstrates that POO (TAG with one palmitoyl and two oleoyls) is the primary TAG form with percentages of 20.18% in A. pernyi and 15.00% in B. mori. The dominating phospholipid species are phosphatidylcholine (PC, 30.40% vs 54.61%) and phosphatidylethanolamine (PE, 34.82% vs 20.39%). Four sterol constituents with total contents of 382.56 ± 3.12 and 371.65 ± 2.98 µg g^?1 are identified and analyzed quantitatively. Additionally, the levels of tocochromanols (20.15 ± 0.89 vs 17.15 ± 0.71 mg g^?1) are quantified in both silkworm pupae oils. Overall, silkworm oil acts as an enriched source of functional lipids and tocochromanols. Practical Applications: A systematic investigation on the principal lipid classes and tocochromanols of Antheraea pernyi pupae and Bombyx mori pupae oil is reported in this study. The informative data provide supporting evidence for comprehensive utilization of silkworm oil for production of nutritional and healthy products.     

Lipid profile of mysid Neomysis intermedia and its changes by feeding DHA-rich marine lipids

There has been no information on lipids of brackish water mysid Neomysis intermedia, which is the most important mysid for human food in Japan. The present study revealed their lipid content, lipid class composition, and fatty acid compositions of total lipids (TL) and major lipid classes. Lipid content of the mysid was 1.0% on wet-weight base. Major lipid classes were phosphatidylcholines (PC, 26%), phosphatidylethanolamines (PE, 24%), and phosphatidylinositols (PI, 14%), whereas the content of triacylglycerols (TAG) was relatively low (7%). Major fatty acids of the TL were 16:0 (21%), 18:1n-9 (12%), 20:5n-3 (EPA; 19%) and 22:6n-3 (DHA; 14%), and the latter two polyunsaturated fatty acids were concentrated in PI (totally 47%) and PE (43%) fractions rather than in PC (20%) and TAG (18%). Arachidonic acid (4%) was also higher in the PI (7%) and PE (5%). When the mysid was fed tri-, di- and monoacylglycerols prepared from DHA-rich fish oil, DHA mainly increased in the TAG fraction of the mysid. The mysid appeared to primarily incorporate exogenous DHA into TAG fraction.     

Regional distribution of tocopherols and fatty acids within soybean seeds

Seed coat, axis, and sections of cotyledons in three soybean cultivars were analyzed by high-performance liquid chromatography for tocopherols, and by gas-liquid chromatography for acyl lipids. Tocopherols were predominantly detected in axis, followed by cotyledons and seed coat. With a few exceptions, dominant components were γ- and δ-tocopherols, with much smaller amounts of α- and β-tocopherols. However, α-tocopherol was higher (P<0.05) for the Mikawajima cultivar than for Okuhara and Tsurunoko in all tissues. Triacylglycerols (TAG) were the major fraction of total lipids, representing 70% in axis and coat and 94% in cotyledons. A small difference (P<0.05) occurred in fatty acid composition of TAG when comparing seed coat to the axis. The fatty acid composition of phosphatidylinositol (PI) differed (P<0.05) from phosphatidylethanolamine (PE) and phosphatidylcholine (PC) in each tissue. Principally, the percentage of palmitic acid was higher, especially in axis and coat. In PE and PC, linoleic was greater, followed by palmitic, in all samples except for seed coat tissue in Mikawajima. The percentages of palmitic acid in both phospholipids were significant higher in the seed coat tissue from this cultivar than in cotyledon or axis of the other varieties. These results suggest that the differences in soybean cultivars could be appreciable, based on the distribution of tocopherols and fatty acids in each component part within soybean seeds.     

Lipids and Fatty Acids of the Gonads of Sea Urchin Diadema setosum (Echinodermata) From the Coastal Area of the Nha Trang Bay,Central Vietnam

The sea urchin Diadema setosum is edible and desirable as food by locals in central Vietnam and a promising target for potential fishing. The lipid profiles of the gonads of the sea urchin inhabiting the coastal area in Nha Trang Bay are studied for the first time. The determination of the content of the total lipids (TL), total phospholipids (PL), monoacylglycerols (MAG), diacylglycerols (DAG), triacylglycerols (TAG), cholesterol (Chol), sterol esters, and free fatty acids (FFA) is analyzed by high-performance thin layer chromatography (HPTLC); the phosphatidylserine (PS), phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidylcholine (PC), lysophosphatidylcholine (LPC), and sphingomyelin (SM) are determined by high-pressure liquid chromatography (HPLC); and the fatty acid levels of TLs are identified using gas chromatography (GC). Non-polar TAG, FFA, Chol, and DAG dominated. The content of total PL is significant. PC is abundant among PL fractions, followed by PE, PI, and PS. The TLs contain a high proportion of PUFA, mainly due to arachidonic fatty acid and eicosapentanoic fatty acid. The (n−6)/(n−3) ratio is 1.68, and the atherogenic and thrombogenic indexes of D. setosum are 1.13 and 0.73. The results can be used in the development of marine bioprospecting and methodological approaches for the creation of functional substances.     

Differential Impact of Sex on Seasonal Changes in Mantle and Tentacle Phospholipids and Triacylglycerols in Sepia officinalis

The aim of the present work was to study the effect of season on phospholipids and triacylglycerols (TAG) of mantle and tentacles of female and male wild Sepia officinalis. The identified phospholipids were phosphatidylethanolamine (PtdEtn), phosphatidylcholine (PtdCho), phosphatidylserine (PtdSer), and phosphatidylinositol (PtdIns), and PtdEtn was the major fraction. Results showed apparent seasonal variation of phospholipid content, particularly with female samples. Fatty acid composition of phospholipid classes showed a differentiation much more in the proportions than in the diversity of fatty acids. Results showed that the major saturated fatty acids were 16:0 and 18:0, the major monounsaturated fatty acids were 18:1 and 20:l, and the major polyunsaturated fatty acids were docosahexaenoic acid (22:6n-3) (DHA) and eicosapentaenoic acid (20:5n-3) (EPA). The results relative to TAG demonstrated significant variations. Principal component analysis confirmed the seasonal and sexual effects. This study could be appropriate for the improvement of consistent monitoring of phospholipid and TAG accumulation in cephalopod, which might be important for both physiological studies and food industries.     

The production potential of eicosapentaenoic and arachidonic acids by the red algaPorphyridium cruentum

The red microalgaPorphyridium cruentum is a new source for eicosapentanoic acid (EPA) and arachidonic acid (AA) fatty acids of potential pharmaceutical value. The conditions leading to a high content of either fatty acid were investigated. The highest EPA content was obtained under conditions resulting in high growth rate (2.4% of ash free dry weight in Strain 1380-1d). High AA content was obtained under slow growth conditions and was maximal in th stationary phase or under nitrogen starvation (2.9%). Strain 1380-la had the highest content (1.9%) of arachidonic acid under exponential growth conditions. By imposing nitrogen starvation, it was possible to obtain a lipid mixture which may be separated into AA and EPA rich fractions.