Sponge fatty acids. 3. Occurrence of series of n−7 monoenoic andiso-5,9 dienoic long-chain fatty acids in the phospholipids of the marine spongeCinachyrella aff.schulzei keller

The fatty acid composition of phospholipids from the New Caledonian spongeCinachyrella aff.schulzei Keller was studied. More than 60 fatty acids were identified as methyl esters andN-acyl pyrrolidides by gas chromatography and gas chromatography/mass spectrometry. Two isoprenoid fatty acids also were shown to be present, namely 4,8,12-trimethyltridecanoic and 5,9,13-trimethyltetradecanoic acids. The unusual 6-tetradecenoic, 6-pentadecenoic, 12-nonadecenoic and 26-methylheptacosanoic (iso-28∶0) acids were found for the first time in sponge phospholipids. A series of six n−7 monoenoic long-chain fatty acids (C₂₃ to C₂₈) were identified, including the rare 16-tricosenoic, 18-pentacosenoic and 21-octacosenoic acids. Fifteen fatty acids possessing the typical 5,9 dienoic moiety accounted for 30% of the total fatty acid mixture. Two new fatty acids were identified, namely 5(Z)-octacosenoic and 27-methyl-5(Z),9(Z)-octacosadienoic (iso-5,9-29∶2). Based on gas chromatography/Fourier transform infrared experiments, the double bonds were assigned the (Z) configuration. For part 2 of this series, see Reference 1.     

Novel Cyclopropane Fatty Acids from the Phospholipids of the Caribbean Sponge <Emphasis Type="Italic">Pseudospongosorites suberitoides</Emphasis>

The cyclopropane fatty acids 17-methyl-trans-4,5-methyleneoctadecanoic acid, 18-methyl-trans-4,5-methylenenonadecanoic acid, and 17-methyl-trans-4,5-methylenenonadecanoic acid were characterized for the first time in nature in the phospholipids (mainly PE, PG and PS) of the hermit-crab sponge Pseudospongosorites suberitoides. Pyrrolidine derivatization was the key in identifying the position of the cyclopropyl and methyl groups in the acyl chains and ¹H NMR was used to determine the trans stereochemistry of the cyclopropane ring. The phospholipids from the sponge also contained an interesting series of iso-anteiso Δ^5,9 fatty acids with chain-lengths between 17 and 21 carbons, with the fatty acids (5Z,9Z)-18-methyl-5,9-nonadecadienoic acid and the (5Z,9Z)-17-methyl-5,9-nonadecadienoic acid being described for the first time in sponges. The anteiso α-methoxylated fatty acid 2-methoxy-12-methyltetradecanoic acid was also identified for the first time in nature in the phospholipids of this interesting marine sponge. The novel cyclopropyl fatty acids could have originated from the phospholipids of a cyanobacterium living in symbiosis with the sponge.     

Novel methoxylated FA from the Caribbean sponge Spheciospongia cuspidifera

The Δ⁶ monoenoic methoxylated FA (6Z)-2-methoxy-6-heptadecenoic acid and (6Z)-2-methoxy-6-octadecenoic acid were identified for the first time in nature in the phospholipids from the uncommon Caribbean sponge Spheciospongia cuspidifera. These findings expand the occurrence of Δ⁶ 2-methoxylated FA to C₁₇−C₁₈ chain lengths and establish a new FA biosynthetic possibility for these marine organisms. The novel methoxylated FA also could have originated from the phospholipids of a bacterium in symbiosis with the sponge.     

Identification of Novel α-Methoxylated Phospholipid Fatty Acids in the Caribbean Sponge <Emphasis Type="Italic">Erylus goffrilleri</Emphasis>

The phospholipid fatty acid composition of the Caribbean sponge Erylus goffrilleri is described for the first time. A total of 70 fatty acids with chain lengths between 13 and 29 carbons were identified in the sponge. Methyl-branched fatty acids predominated in E. goffrilleri suggesting the presence of a considerable number of bacterial symbionts. The novel fatty acids (5Z,9Z)-2-methoxy-5,9-hexadecadienoic acid, (5Z,9Z)-2-methoxy-5,9-octadecadienoic acid, (5Z,9Z)-2-methoxy-5,9-nonadecadienoic acid, and (5Z,9Z)-2-methoxy-5,9-eicosadienoic acid are described for the first time in the literature. In addition, the iso-methyl-branched fatty acids (9Z)-2-methoxy-15-methyl-9-hexadecenoic acid and (5Z,9Z)-2-methoxy-15-methyl-5,9-hexadecadienoic acid, also identified in E. goffrilleri, were identified for the first time in nature. Based on the identified metabolites it is proposed that the unprecedented biosynthetic sequence: i-17:1Δ9 → 2-OMe-i-17:1Δ9 → 2-OMe-i-17:2Δ5,9 might be responsible for the biosynthesis of the novel iso-α-methoxylated fatty acids in E. goffrilleri.     

Novel brominated phospholipid fatty acids from the Caribbean spongeAgelas sp.

The very long-chain fatty acids, (5E,9Z)-6-bromo-5,9-tetracosadienoic, (5E,9Z)-6-bromo-23-methyl-5,9-tetracosadienoic, (5E,9Z)-6-bromo-5,9-pentacosadienoic and (5E,9Z)-6-bromo-24-methyl-5,9-pentacosadienoic acids, were identified in the phospholipids (mainly phosphatidylethanolamine) of the spongeAgelas sp. Structure elucidation was accomplished by means of mass spectrometry and chemical transformations, including deuteration with Wilkinson's catalyst. All of the sterols from the sponge had the Δ5,7 nucleus, with 24-methylcholesta-5,7,22-trien-3β-ol (ergosterol) and 24-ethylcholesta-5,7,22-trien-3β-ol being the most abundant.     

The distribution of brominated long-chain fatty acids in sponge and symbiont cell types from the tropical marine spongeAmphimedon terpenensis

The tropical marine spongeAmphimedon terpenensis (family Niphatidae, order Haplosclerida) has previously been shown to possess unusual lipids, including unusual fatty acids. The biosynthetic origin of these fatty acids is of interest as the sponge supports a significant population of eubacterial and cyanobacterial symbionts. The total fatty acid composition of the sponge was analyzed by gas chromatography/mass spectrometry of the methyl esters. Among the most abundant of the fatty acids in intact tissue were 16∶0, 18∶0 and 3,7,11,15-tetramethylhexadecanoic (phytanic) acid. In addition, three brominated fatty acids, (5E,9Z)-6-bromo-5,9-tetracosadienoic acid (24∶2Br), (5E,9Z)-6-bromo-5,9-pentacosadienoic acid (25∶2Br) and (5E,9Z)-6-bromo-5,9-hexacosadienoic acid (26∶2Br) were also present. The three brominated fatty acids, together with phytanic acid, were isolated from both ectosomoal (superficial) and choanosomal (internal) regions of the sponge. Analysis of extracts prepared from sponge/symbiont cells, partitioned by density gradient centrifugation on Ficoll, indicated that phytanic acid and the three brominated fatty acids were associated with sponge cells only. Further, a fatty acid methyl ester sample from intact tissue ofA. terpenensis was partitioned according to phospholipid class, and the brominated fatty acids were shown to be associated with the phosphatidylserine and phosphatidylethanolamine fractions that are commonly present in marine sponge lipids. The phosphatidylcholine and phosphatidylglycerol fractions were rich in the relatively shorter chain fatty acids (16∶0 and 18∶0). The association of brominated long-chain fatty acids (LCFA) with sponge cells has been confirmed. The findings allow comment on the use of fatty acid profiles in chemotaxonomy and permit further interpretation of LCFA biosynthetic pathways in sponges. The assignment of the sponge studied, which is currently placed asA. terpenensis, is being supported to some extent, but the species is unusual in having C₂₅ fatty acids as the major constituent in this group. Other factors, such as season or microenvironmental conditions, may influence observed fatty acid composition which tends to reduce the usefulness of fatty acid profiles as markers in sponge chemotaxonomy.     

Long chain fatty acid deficits in brain myelin sphingolipids of undernourished rat pups

A restricted maternal dietary intake (40% of ad libitum intake) is known to cause myelin deficit that is accompanied by decreased amounts of individual phospholipids and sphingolipids in brain myelin of suckling rats. This communication reports the effects of the same nutritional stress on the fatty acid composition of brain myelin lipids. In myelin of 19-day-old normally fed rats, palmitate (16∶0), stearate (18∶0) and oleate (18∶1) accounted for 80–90% of all fatty acids in phosphatidylcholine, phosphatidylethanolamine and phosphatidylserine. Maternal dietary restriction resulted in deficits of total fatty acid content, but did not affect the proportional distribution of individual fatty acids among phospholipids. By contrast, longer chain (22- and 24-carbon) fatty acids accounted for more than half the fatty acid content of myelin cerebroside and sulfatide from the 19-day-old control rat pups. In undernourished rats of that age, proportions of lignocerate (24∶0) and nervonate (24∶1) in cerebroside and sulfatide were 40–50% lower than those in control rats. This, together with higher proportions of 16∶0, 18∶0 and 18∶1 and a higher ratio of C₁₆−C₂₀ to C₂₂−C₂₄ in under-nourished than in control rats, suggests an impairment in fatty acid chain elongation. Ten days of nutritional rehabilitation failed to restore the fatty acid imbalances; however, after an additional 5 days of ad libitum feeding, the experimental and control values were similar. The undernutrition results in hypomyelination, which is characterized by a proportional decrease in lignoceric and nervonic acids of sphingolipids.     

Phospholpid studies of marine organisms: 2.1 Phospholipids,phospholipid-bound fatty acids and free sterols of the spongeAplysina fistularis (Pallas) formafulva (Pallas) (=Verongia thiona)2. Isolation and structure elucidation of unprecedented branched fatty acids

The free sterols and phospholipids of the demospongeAplysina fistularis were isolated and analyzed. The free sterols consisted mainly of the unusual 26-methylated sterols aplysterol (53%) and 24(28)-dehydroaplysterol (7%) together with 7 commonly occurring sterods. The major phospholipids were phosphatidylcholine, phosphatidylglycerol, phosphatidylinositol, phosphatidylethanolamine, phosphatidylserine and diphosphatidylglycerol. The major fatty acyl components of the phospholipids consisted of 85% C₁₄−C₂₀ acids, including the unprecedented 2,6,10-trimethyl-5-tetradecenoic acid and 11-methyloctadecanoic acid. The remaining 15% were C₂₇−C₃₀ demospongic acids, including 2 novel acids tentatively assigned the structures 5,9,23-octacosatrienoic acid and 5,9,23-nonacosatrienoic acid, and 3 novel acids proven to be 5,9,21-octacosatrienoic acid, Z,Z-20-methyl-5,9-hexacosadienoic acid and Z,Z-22-methyl-5,9-octacosadienoic acid. The biosyntheses of the novel demospongic acids are proposed to occur by chain elongation of monoenoic or branched precursors followed by desaturation. The large quantities of typically bacterial phospholipids and fatty acids found implied the presence of bacteria in the sponge, in agreement with microscopic studies. Analysis of the phospholipid-bound fatty acids in a sponge cell-enriched fraction indicated that the demospongic acids, including the 2 branched structures, were the major acids of the sponge cells. The presence inA. fistularis of demospongic acids containing membrane disordering groups—methyl branches or double bonds—on the ω7 carbon is proposed to be due to the need by the sponge for membranes possessing fluidity near the middle of the phospholipid bilayer. It is also proposed that the C₂₆ methyl group of aplysterol causes disordering of the phospholipid bilayer in the same region, and thus also evolved in response to this need. For part 1, see ref. 1. Phylum porifera, class demospongia, subclass ceractinomorpha, order verongida, family aplysinidae (2). Formerly known asVerongia thiona de Laubenfels. Major diagnostic fragments from peak 23:M⁺ 471 (2.9%); m/z 442 (C₂₆, 0.2%); m/z 428 (C₂₅, 0.2%); m/z 414 (C₂₄, 0.1%); m/z 400 (C₂₃, 0.3%); (absence of C₂₂ peak); m/z 374 (C₂₁, 0.2%); m/z 360 (C₂₀, 0.2%); m/z 234 (C₁₁, 1.0%); m/z 220 (C₁₀, 0.7%); m/z 206 (C₉, 0.5%); m/z 194 (C₈, 0.4%); m/z 180 (C₇, 24.9%); m/z 166 (C₆, 1.2%); m/z 152 (C₅, 0.8%); m/z 140 (C₄, 1.3%); m/z 126 (C₃, 18.7%). Major diagnostic fragments from peak 25:M⁺ 485 (0.9%); m/z 456 (C₂₇, 0.1%); (absence of C₂₆, C₂₅ peaks); m/z 414 (C₂₄, 0.3%); m/z 400 (C₂₃, 0.1%); m/z 402 (C₂₃, 0.1%); (absence of C₂₂ peak); m/z 374 (C₂₁, 0.1%); m/z 360 (C₂₀, 0.1%); m/z 234 (C₁₁, 0.5%); m/z 220 (C₁₀, 0.3%); m/z 206 (C₉, 0.1%); m/z 194 (C₈, 0.2%); m/z 180 (C₇, 18.0%); m/z 166 (C₆); m/z 152 (C₅), m/z 140 (C₄); m/z 126 (C₃, 15.0%).     

Characterization of skin-surface lipids from the monkey (Macaca fascicularis)

Skin-surface lipids from the monkeyMacaca fascicularis are composed of sterol esters (38%), cholesterol (4%) and two types of wax diesters, identified as Type II (IIa and IIb, 17% and 40%, respectively). Type IIa contained diesters of 1,2-alkanediols esterified with two molecules of long-chain (C₁₄−C₃₄) fatty acids having straight and branched chains. In the diesters IIa, fatty acids shorter than C₁₉ predominated in position 1, and fatty acids longer than C₂₀ predominated in position 2. Type IIb contained diesters of 1,2-alkanediols esterified with C₄ and C₅ branched-chain fatty acids (predominantly isovaleric acid) at position 1 and long-chain (C₁₄−C₂₇) acids, having straight and branched chains, at position 2. The shortchain acids were converted to 2-nitrophenylhydrazides and analyzed by high-performance liquid chromatography (HPLC). Ammonia chemical ionization (CI)-gas chromatography (GC)-mass spectrometry (MS) resolved the intact diesters IIb into 12 peaks corresponding to molecular weights ranging from 597 to 748, and showed that the molecular species, such as C₂₁−C₁₆−C₅ (diol, fatty acid in position 2, fatty acid in position 1), C₂₂−C₁₆−C₅ and C₂₃−C₁₆−C₅, were prevalent. The fatty acids from both diesters were mostly (>98%) saturated. The 1,2-alkanediols from both diesters consisted of C₁₆−C₂₆ saturated straight- and branched-chain components. The acyl groups of sterol esters contained 86% C₁₄−C₃₄ branched-chain acids. The unsaturated fatty acids (5.4%) belonged to a straight-chain monoenoic series having extremely long chains (C₁₈−C₃₄). The branched-chain structures in the fatty acids and diols were iso and anteiso. These results show the species-specific profile for the skin-surface lipid synthesis.     

The fatty acid composition of a Vibrio alginolyticus associated with the alga Cladophora coelothrix. Identification of the novel 9-methyl-10-hexadecenoic acid

The fatty acid composition of a new strain of Vibrio alginolyticus, found in the alga Cladophora coelothrix, was studied. Among 38 different fatty acids, a new fatty acid, 9-methyl-10-hexadecenoic acid and the unusual 11-methyl-12-octadecenoic acid, were identified. Linear alkylbenzene fatty acids, such as 10-phenyldecanoic acid, 12-phenyldodecanoic acid and 14-phenyltetradecanoic acid, were also found in V. alginolyticus. The alga contained 43% saturated fatty acids, and 28% C₁₆–C₂₀ polyunsaturated fatty acids of the n−3 and n−6 families.     

Identification of naturally occurringtrans,trans Δ5,9 fatty acids from the spongePlakortis halichondroides

The firsttrans fatty acids from a sponge have been isolated fromPlakortis halichondroides. The sponge was shown to contain theiso acids (5E,9E)-19-methyl-5,9-eicosadienoic, (5E,9E)-20-methyl-5,9-heneicosadienoic and (5E,9E)-21-methyl-5,9-docosadienoic acid, as well as theanteiso acids (5E,9E)-19-methyl-5,9-heneicosadienoic and (5E,9E)-20-methyl-5,9-docosadienoic acid together with the straight chain (5E,9E)-5,9-docosadienoic acid. The acids were shown by gas chromatography, Fourier transform infrared and¹³C nuclear magnetic resonance to contain thetrans configuration. An eicosadienoic acid, namely (6E,14E)-6,14-eicosadienoic acid, and 12-methyl-5-octadecenoic acid were also identified in a sponge for the first time. The fatty acids were shown to be the principal constituents of phosphatidylethanolamine and phosphatidylcholine. No sterols were found in the sponge. The results presented in this work should be helpful in taxonomy of Homoscleromorpha.     

Production of odd chain polyunsaturated fatty acids byMortierella fungi

A soil isolate,Mortierella alpina 1S-4, was found to show high production of odd chain polyunsaturated fatty acids (PUFAs) among various arachidonic acid-producingMortierella strains tested. The fungus mainly accumulated 5,8,11,M-cis-nonadecatetraenoic acid. With 5%n-hepta-decane and 1% yeast extract as growth substrates, the amount of C_19:4:4 acid accumulated reached 44.4 mg/g dry mycelia (0.68 mg/mL of culture broth). This value accounted for 11.2% of the total fatty acids in the extracted lipids from mycelia, and odd chain fatty acids comprised over 95% of the total mycelial fatty acids. The addition of sesamin, a specific inhibitor of A5 desaturation, caused an increase in C_19:3 acid and an accompanying decrease in C_19:4 acid. On the other hand, species ofMortierella that could not produce C-20 PUFAs accumulated C-17 acids, but no C-19 PUFAs, when grown with fatty substrates with an odd chain skeleton. The odd chain PUFAs were distributed in both neutral and polar lipids. The biosynthetic route to C_19:4 acid was presumed to mimic the n-6 route to arachidonic acid as follows: C_17:0 → C_17:1→ C_17:2→ C_17:3 → C_19:3 → C_19:4 acids. On leave from Suntory Ltd.     

The effect of a fat free diet on esterified monoenoic fatty acid isomers in rat tissues

Rats were maintained 120–140 days on a normal diet (group 1) or one deficient in fatty acids (group 2). Isomer composition was determined of monoenoic fatty acids (16∶1, 18∶1) isolated from total lipids of heart, kidney, lung, brain and lumbar fat, and from separated neutral lipids and phospholipids of heart and kidney. Group 1: The number of major isomers of C_16∶1 and C_18∶1 was similar in all tissues but their proportions varied in different tissues and types of lipid. Group 2: The proportions of 16∶1(n−7) increased and of other 16∶1 isomers decreased in all tissues; 18∶1(n−9) was increased at the expense of (n−7) in heart, to a lesser extent in kidney, and was little changed in lung, lumbar fat, or brain. The decrease in proportion of 18∶1(n−7) was greatest in heart-muscle phospholipids. C_20∶3 comprised 95% (n−9) and 5% (n−7) in heart and kidney lipids. The changes in group 2 probably represent the body’s attempts to maintain lipids with the physical and chemical properties necessary to normal biological function. Nomenclature of fatty acids as in IUPAC-IUB Commission on Biochemical Nomenclature, “The nomenclature of lipids,” J. Biol. Chem. 242:4845–49 (1967).     

New 2-Methyl-13-Icosenoic Acid from the Temperate Calcisponge <Emphasis Type="Italic">Leuconia johnstoni</Emphasis>

The fatty acid composition of the temperate calcareous marine sponge Leuconia johnstoni Carter 1871 (Calcaronea, Calcarea) was characterized for the first time in specimens collected off the Brittany coast of France over four years from October 2005 to September 2008. Forty-one fatty acids (FA) with chain lengths ranging from C₁₄ to C₂₂ were identified as fatty methyl esters (FAME) and N-acyl pyrrolidide (NAP) derivatives by gas chromatography–mass spectrometry (GC–MS). Twenty-two saturated fatty acids (SFA) were identified accounting for 52.1–59.0% of the total FA and dimethylacetals (DMA). In addition, among the SFA, we noticed the presence of numerous methyl-branched iso and anteiso FA, suggesting a large number of associated bacteria within L. johnstoni. Thirteen monounsaturated fatty acids (MUFA, 28.0–36.0% of total FA + DMA) were also identified as well as six polyunsaturated fatty acids (PUFA, 4.0–8.2%). A noticeable DMA was detected at a high level, particularly in September 2008 (11.8%), indicating the presence of plasmalogens in this sponge species. This calcareous sponge lacked the non-methylene-interrupted FA (NMI FA) with a Δ5,9 system typical of siliceous Demosponges and Hexactinellids. The occurrence of the unusual 8,13-octadecadienoic acid was reported for the first time as a minor PUFA in a calcareous sponge. The major FA, representing 20–25% of this sponge FA, was identified as the new 2-methyl-13-icosenoic acid from mass spectra of its methyl ester and its corresponding N-acyl pyrrolidide derivatives as well as a dimethyl disulfide adduct.     

The steryl ester and phospholipid fatty acids of the spongeAgelas conifera from the Colombian Caribbean

The steryl ester and phospholipid fractions of the marine spongeAgelas conifera were isolated and analyzed. The fatty acyl components of the steryl ester and phospholipid fractions as determined by gas chromatography and gas chromatography/mass spectrometry were very similar and consisted of 56.8 and 62.7% of C₁₄−C₂₀ acids (normal; branched, especiallyiso andanteiso; and monounsaturated, particularly Δ⁹ and Δ¹¹ acids) and of 43.1 and 35.5% of C₂₄−C₂₆ acids (Δ^5,9 diunsaturated acids), respectively. The major constituent fatty acids detected were 13-methyltetradecanoic,n-hexadecanoic, 10-methylhexadecanoic, 11-octadecenoic, 12-methyloctadecanoic, 5,9-pentacosadienoic and 5,9-hexacosadienoic acids. The phospholipids isolated were identified as phosphatidylcholine (37%), phosphatidylserine (34%), phosphatidylethanolamine (16%) and phosphatidylinositol (11%). The distribution of fatty acids within the phospholipid classes was also determined.     

Isolation and characterization of the monounsaturated long chain fatty acids ofMycobacterium tuberculosis

The positions of double bond in the monounsaturated C₁₅−C₃₂ fatty acids ofMycobacterium tuberculosis H37Ra were established by gas chromatography/mass spectrometry of the ozonized esters and their pyrrolidide derivatives. The monounsaturated C₁₅−C₂₁ fatty acids had the double bond primarily at the Δ⁹ position while the monounsaturated longer chain fatty acids (C₂₂−C₃₂) had the double bond in several positions. Many of the latter acids, especially the odd-numbered series, were very complex isomeric mixtures. Quantitation showed the most abundant even-numbered long chain fatty acid isomers to be as follow: C₂₂, Δ⁴; C₂₄, Δ⁵; C₂₆, Δ⁷ and Δ⁹; C₂₈, Δ⁹; C₃₀, Δ¹¹ and Δ¹³; C₃₂, Δ¹³ and Δ¹⁵.     

Monoenoic fatty acids in human brain lipids: Isomer identification and distribution

The carbon chain length distribution and the double bond positional isomer composition of the monoenoic fatty acids of the lipids of total human brain tissue have been determined using gas chromatography and gas chromatography/mass spectrometry of the fatty acid methyl and picolinyl esters. The even chain length monoenoic C₁₆ to C₂₈ fatty acids contain predominantly two positional isomer series, the n−7 and n−9cis homologues, whose relative proportion varies significantly with chain length. The odd chain length long-chain fatty acids consist of n−8 and n−10 isomers, whereas the odd chain length very long-chain (more than 22 carbon) fatty acids are n−7 and n−9 isomers.     

Positional analysis of triglycerides and phospholipids rich in long-chain polyunsaturated fatty acids

Four sources of long-chain polyunsaturated fatty acids (LCP) differing in their chemical structure (triglycerides or phospholipids) and in their origin (tuna triglycerides, fungal triglycerides, egg phospholipids, and pig brain phospholipids) were analyzed to determine the distribution of the component fatty acids within the molecule. Lipase and phospholipase A₂ hydrolysis was performed to obtain 2-monoacylglycerols and lysophospholipids, respectively, which allowed us to determine the distribution of fatty acids between the sn-2 and sn-1,3 positions of triglycerides or between the sn-1 and sn-2 position of phospholipids. Fatty acids in the LCP sources analyzed were not randomly distributed. In tuna triglycerides, half of the total amount of 22∶6n−3 was located at the sn-2 position (49.52%). In fungal triglycerides, 16∶0 and 18∶0 were esterified to the sn-1,3 (92.22% and 91.91%, respectively) 18∶1 and 18∶2 to the sn-2 position (59.77% and 62.62%, respectively), and 45% of 20∶3n−6 and only 21.64% of 20∶4n−6 were found at the sn-2 position. In the lipid sources containing phospholipids, LCP were mainly esterified to the phosphatidylethanolamine fraction. In egg phospholipids, most of 20∶4n−6 (5.50%, sn-2 vs. 0.91%, sn-1) and 22∶6n−3 (2.89 vs. 0.28%) were located at the sn-2 position. In pig brain phospholipids, 22∶6n−3 was also esterified to the sn-2 (13.20 vs. 0.27%), whereas 20∶4n−6 was distributed between the two positions (12.35 vs. 5.86%). These results show a different fatty acid composition and distribution of dietary LCP sources, which may affect the absorption, distribution, and tissue uptake of LCP, and should be taken into account when supplementing infant formulas.     

New 17-Methyl-13-Octadecenoic and 3,16-Docosadienoic Acids from the Sponge <Emphasis Type="Italic">Polymastia penicillus</Emphasis>

The phospholipid fatty acid composition of the North-East Atlantic sponge Polymastia penicillus (South Brittany, France) was investigated. Sixty fatty acids (FA) were identified as methyl esters (FAME) and N-acyl pyrrolidides (NAP) by gas chromatography–mass spectrometry (GC/MS), including eight Δ5,9 unsaturated FA and three long-chain 2-hydroxylated FA. The major phospholipid FA were palmitic (14.3% of the total FA mixture), vaccenic (12.7%), 15(Z)-docosenoic (13.4%) and 5(Z),9(Z)-hexacosadienoic (13.3%) acids. In addition to the iso- and anteiso-branched saturated FA, several unusual short-chain branched saturated FA were identified. In addition to the known Δ5,9 FA, and interestingly regarding their identification by GC–MS as N-acyl pyrrolidides, was the co-occurrence of unusual FA possessing a Δ3, Δ4 and Δ5 double bond such as iso-4-pentadecenoic, iso-5-heptadecenoic, anteiso-5-heptadecenoic and two new compounds, not hitherto found in nature, namely 17-methyl-13-octadecenoic (0.8%) and 3,16-docosadienoic (1.1%) acids.     

A new cytotoxic fatty acid (5<Emphasis Type="Italic">Z</Emphasis>,9<Emphasis Type="Italic">Z</Emphasis>)-22-methyl-5,9-tetracosadienoic acid and the sterols from the far Eastern sponge <Emphasis Type="Italic">Geodinella robusta</Emphasis>

A new fatty acid, (5Z,9Z)-22-methyl-5,9-tetracosadienoic acid (1a), and a rare fatty acid, (5Z,9Z)-23-methyl-5,9-tetracosadienoic acid (2a), the predominant constituents of the free fatty acid fraction from the lipids of the sponge Geodinella robusta, were isolated and partly separated by reversed phase high-performance liquid chromatography, followed by multifold crystallization from MeOH to give 1a and 2a in 70% and 60% purity, respectively. These fatty acids were identified as (5Z,9Z)-22-and (5Z,9Z)-23-methyl-5,9-tetracosadienoic acids by nuclear magnetic resonance techniques, including distortionless enhancement by polarization transfer, heteronuclear multiple quantum connectivity, and correlation spectroscopy experiments, as well as from mass-spectrometric data for their methyl esters, the methyl esters of their perhydro derivatives, and their pyrrolidides. Mixtures of 1a and 2a showed cytotoxic activity against mouse Ehrlich carcinoma cells and a hemolytic effect on mouse erythrocytes. The sterol fraction from the same sponge was analyzed by gas liquid chromatography mass spectrometry, and 24-methylenecholesterol was identified as a main constituent of this fraction. The implications of the co-occurrence of membranolytic long-chain fatty acids and 24-methylenecholesterol as a main membrane sterol are discussed in terms of the phenomenon of biochemical coordination.