Positional distribution of fatty acids in depot triglycerides of aquatic animals

Stereospecific triglyceride analyses were performed on fats of the following animals: five aquatic invertebrates, five freshwater fish, six marine fish, three marine birds, two amphibia, two seals, a whale, and a marine turtle. The distribution of faty acids was asymmetrical in most cases. A formula is presented which describes the general tendencies of fatty acid distribution in many animal fats, and some special rules which modify this formula are stated. Visit sponsored by a grant from the National Institutes of Health (AM-06011).     

Reinvestigation of positional distribution of fatty acids in docosahexaenoic acid-rich fish oil triacyl-sn-glycerols

Positional distribution of fatty acids in triacyl-sn-glycerols of docosahexaenoic acid (DHA)-rich tuna orbital and bonito head oils has been reanalyzed by a method based on chromatographic separation of isomeric and enantiomeric monoacyl-sn-glycerol (MAG) derivatives. When boric acid thinlayer chromatography (TLC) was used for separation of 1(3)- and 2-MAG analytical intermediates, the stereospecific analysis showed the preferential association of DHA to the sn-2 position followed by the sn-3 position. This distribution pattern differed from that obtained by silicic acid LTC of their bis-3,5-dinitrophenylurethane (DNPU) derivatives. Reversed-phase high-performance liquid chromatography elution profiles of 1(3)- and 2-MAG intermediates revealed that 1(3)- and 2-MAG made up of both short- and long-chain lengths cannot be clearly resolved by TLC after preparation of the DNPU derivatives. The 1(3)- and 2-MAG must be resolved by boric acid TLC prior to derivatization.     

Production of docosahexaenoic and docosapentaenoic acids bySchizochytrium sp. isolated from Yap Islands

A marine microbe (strain SR21) from the coral reef area of the Yap Islands was isolated by a screening test for polyunsaturated fatty acids and was found to accumulate lipid that contained 22:5n-6 docosapentaenoic acid (DPA) as well as 22:6n-3 docosahexaenoic acid (DHA). Strain SR21 was identified as genusSchizochytrium in Labyrinthulomycota, owing to its ultrastructural character and life cycle, which is composed of vegetative cell, zoosporangium, and zoospore stages. After cultural optimization, both in flask and fermenter, the highest DHA and DPA productivities of 2.0 and 0.44 g/L per day, respectively, were obtained in a medium of 60 g/L glucose and corn steep liquor/ammonium sulfate in a half salt concentration of seawater in fermenter culture at 28°C and pH 4. This productivity was almost twice that obtained with flask culture, indicating its high resistance to mechanical stirring. The lipid extracted from the cell was about 50% of the dry cell weight and was composed of 93% triacylglycerol (TG). DHA content of the lipid was 34% of total fatty acids. The TG profile was simple, and the content of the most dominant TG, 1,3-dipalmitoyl-2-DHA-TG, was 27%. TG that contained DHA and n-6-DPA amounted to 57 and 17%, respectively, of total TG molecules. Strain SR21 was revealed to be an excellent source of microbial DHA and n-6 DPA.     

The positional distribution of fatty acids in the phospholipids and triglycerides ofMycobacterium smegmatis andM. bovis BCG

Position 1 of the phospholipid and triglyceride fractions isolated fromMycobacterium smegmatis andM. bovis BCG was esterified principally with C₁₈ related fatty acids (18∶0, 18∶1 and 19Br). Position 2 was occupied principally by C₁₆ fatty acids. The third position of the triglycerides was esterified with a preponderance of C₂₀+fatty acids. Seventysix per cent of position 3 fatty acids in BCG and 43% inM. smegmatis triglycerides contained fatty acids of greater than 20 carbon atoms.     

Fatty acid positional distribution in egg yolk triglycerides from various avain species

The fatty acid composition and distribution in egg yolk triglycerides and phosphatides from the turkey, duck, prairie chicken, bobwhite quail, Japanese quail, and inbred-hybrid and midget mutant hens were determined after all species had been fed diets of similar fat and fatty acid content for 90 days. Total volk lipids were composed of ca. two-thirds neutral lipids and one-third polar lipids. The predominant fatty acids were palmitic and stearic. There were statistically significant differences in the my ristic, palmitic, palmitoleic, linoleic, and linolenic acids in the yolk triglycerides and in the proportion of 16∶1, 18∶0, 18∶2, arachidonic, docosanoic, docosahexaenoic, and tetracosanoic acids in the phosphatides among the various species. Linoleic acid predominantly was linked at the 2-position in the yolk triglycerides followed by the 20∶4 acid. The 18∶1 acid also was found preferentially at the 2-position. There was a low level of 18∶2 in the yolk triglycerides and phosphatides from the duck and an especially high level of 20∶4 acid in the phosphatides. The triglycerides in the species studied have essentially the same distribution of fatty acids in the 2-position. In all the species, the affinity for the fatty acids at the 2-position is in the following order: 18∶2=20∶4>18∶1 =18∶3>18∶0=16∶1>14∶0>16∶0 Differences observed among the various genera did not appear to follow taxonomic boundaries. The duck has an efficient system for converting 18∶2 into 20∶4 by elongation and desaturation. The prairie chicken apparently has a high requirement for 18∶2 but an inadequate system for its conversion into 20∶4.     

Determination of the positional distribution of fatty acids in butterfat triacylglycerols

Triacylglycerol (TAG) standards were separated by analytical high-performance liquid chromatography (HPLC) with laser light-scattering detection (LLSD). A high sensitivity for TAGs was observed with LLSD whereas poor sensitivity was observed with ultraviolet detection. The HPLC-LLSD analytical separation of butterfat TAGs showed that the TAGs were eluted according to increasing carbon number. Preparative HPLC-LLSD was used to characterize butterfat TAGs that contained hypercholesterolemic fatty acids (laurate, myristate, palmitate) with carbon chainlengths of 12 or greater. These TAG fractions accounted for 29.2% of the total butterfat TAGs. Analysis of the positional distribution of fatty acids of selected butterfat TAGs containing hypercholesterolemic fatty acids showed the presence of positional isomers in each of these fractions. These butterfat TAGs also showed the predominant presence of hypercholesterolemic fatty acids at thesn-2 position. The characterization of the positional distribution of hypercholesterolemic fatty acids in butterfat TAGs is the first step for the determination of the metabolic role of the positional distribution in the hypercholesterolemic effects of butter.     

A comparison of the positional distribution of fatty acids in milk triglycerides of the extant monotremes platypus (Ornithorhynchus anatinus) and echidna (Tachyglossus aculeatus)

Milk triglycerides from the platypus were subjected to fatty acid and stereospecific analysis to determine the positional distribution of fatty acids in the triglycerides. Of the major fatty acids, 12∶0 was preferentially esterified at thesn-3 position, 14∶0 and 16∶0 were selectively associated with thesn-2 position, and 18∶0 was located predominantly at thesn-1 position. The unsaturated fatty acids, 14∶1, 16∶1, 18∶1, 18∶2 and 18∶3, were preferentially esterified at thesn-3 position. The fatty acid distribution pattern of the platypus, a monotreme, is similar to that of marsupials and eutherians but is in contrast to the only other extant monotreme, the echidna.     

Specific distribution of fatty acids in the milk fat triglycerides of goat and sheep

The triglycerides of the fat globules of sheep and goat milk were isolated and separated into short and long chain lengths by silicic acid column chromatography. The short chain lengths comprised major triglycerides with 34–44 acyl carbon atoms and accounted for nearly 50% of the total milk fat. The long chain lengths contained major triglycerides with 40–54 acyl carbons. Stereospecific analyses of the short chain triglyceride fraction showed that of the 20–23 moles per cent of C₄−C₈ fatty acids present, at least 95% were specifically attached to the glycerol molecule in the position corresponding to carbon 3 ofsn-glycerol. The distribution of the other fatty acids (C₁₀ or greater) did not show such marked specificity for either the 1 or the 2 position. Although individual triglycerides were not identified, the specific placement of the fatty acids could best the accounted for by assuming a common pool of long chain 1,2-diglycerides which served as precursors of the bulk of both short and long chain triglycerides during milk fat synthesis. Presented in part at the AOCS Meeting, New York, October 1968.     

Composition and positional distribution of fatty acids in phospholipids isolated from pork muscle tissues

The composition and positional distribution of fatty acids in phospholipids isolated from four locations of a hog carcass is presented. Variations in fatty acid composition of phospholipids were found depending upon the location in the carcass. The total unsaturated fatty acid content averaged 34.3 mole % for lecithin, 52.5 mole % for phosphatidylethanolamine, 40.3 mole % for phosphatidylserine and 41.3 mole % in sphingomyelin. The cephalins had a much higher percentage of polyunsaturated fatty acids than lecithin. The chief saturated fatty acid in lecithin and sphingomyelin was palmitic and in cephalins it was stearic. A snake venom enzyme preparation(Crotalus adamanteus) hydrolyzed primarily unsaturated fatty acids in phosphoglycerides and the higher the percentage of unsaturation within the fatty acid the higher percentage of hydrolysis occurred. The unsaturated fatty acids were found chiefly at the theβ-position and the saturated fatty acids at thea-position in the phosphoglycerides. Michigan State Agricultural Experiment Station Publication No. 3389. Supported by the U.S. Public Health Service Research Grant No. GM 08801-03.     

Effects of long-term feeding of marine oils with different positional distribution of eicosapentaenoic and docosahexaenoic acids on lipid metabolism, eicosanoid production, and platelet aggregation in hypercholesterolemic rats

Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) were distributed mainly in the sn-1,3 positions of seal oil triglyceride and in the sn-2 position of squid oil triglyceride. Seal oil-rich or squid oil-rich fats having constant saturated/monounsaturated/polyunsaturated fatty acid (PUFA) and n−6/n−3 PUFA ratios were fed to exogenously hypercholesterolemic rats for 160 d. The control fat contained linoleic acid as the sole PUFA. Before starting the experimental diets, rats were orally treated with high doses of vitamin D for 4 d to accelerate atherogenesis. The percentage of arachidonic acid in phosphatidylcholine and phosphatidylethanolamine of liver, platelets, and aorta was lower in the marine oil groups than in the control group, seal oil being more effective than squid oil. Maximal platelet aggregation induced by collagen was significantly lower both marine oil groups. Platelet thromboxane (TX) A₂ production induced by collagen or thrombin was markedly reduced by feeding seal or squid oils, the reduction being more pronounced in the seal oil than in the squid oil group. Aortic prostacyclin (PGI₂) production was the same among the three groups. The ratio of the productions of aortic PGI₂ and platelet TXA₂ was significantly higher in the seal oil than in the control group. Although there was no difference in intimal thickness among the three groups, the aortic cholesterol content was significantly lower in the marine oil groups than in the control group. These results showed that the main effects in rats of the different intramolecular distributions of EPA and DHA in dietary fats were on arachidonic acid content in tissue phospholipids and on platelet TXA₂ production.     

Polymorphism in single-acid triglycerides of positional and geometric isomers of octadecenoic acid

The polymorphism of 25 glycerol trioctadecenoates with double bonds ranging from Δ4-Δ17 was investigated by differential scanning calorimetry. Triglycerides withcis bonds in odd positions Δ7-Δ13 exhibited three intermediate melting (β′-) forms, but those withcis bonds in even positions, exceptcis Δ4, lacked β′-forms. Among thetrans compounds, only Δ11, 13, and 14 showed β′-forms. Thecis andtrans Δ5 triglycerides were unusual, because they readily assumed low melting (α-) forms that were not easily converted to high melting (β-) forms. β-Form mp of compounds in each series (cis ortrans) alternated depending upon double bond position; an even position correlated with high mp. Heats of fusion (ΔH_f) for β-forms, likewise, fluctuated with double bond position but nonuniformly;trans Δ6 had the highest ΔH_f (43 cal/g),cis Δ12 the lowest (21 cal/g).     

Positional distribution of the fatty acids in the triglycerides of the oil of some african peanut varieties

The distribution of fatty acids between the sn-1-, sn-2- and sn-3-positions of the triglycerides from the oils of eight African peanut varieties has been determined. The saturated fatty acids and eicosenoic acid occur almost exclusively at the sn-1- and sn-3-positions. The sn-1-position contained slightly more palmitic acid than the sn-3-position. The fatty acids with a chain length exceeding 18 carbons were preferentially incorporated in the sn-3-position. Linoleic acid was preferentially esterified at the sn-2-position, whereas oleic acid was equally distributed among the three positions. The amount of the saturated fatty acids, i.e., palmitic and stearic acid, and of oleic acid and linoleic acid incorporated in the sn-1-, sn-2-and sn-3-position, were each linearly related to their respective content in the triglycerides.     

Positional distribution of fatty acids in triglycerides from milk of several species of mammals

Milk triglycerides from the echidna, koala, Tammar wallaby, guinea pig, dog, cat, Weddell seal, horse, pig and cow were subjected to fatty acid and stereospecific analysis to determine the positional distribution of the fatty acids in the triglycerides. The samples presented a wide range of fatty acids, most of which varied in content among species. The compositions of the acids at the 3 positions also varied among species, reflecting the content of these acids in the triglycerides. However, there was a general similarity in fatty acid positional distribution patterns for all the species with the exception of the echidna. The echidna exhibited a completely different fatty acid positional distribution pattern. The saturated acids were preferentially esterified at thesn-1-position whereas the unsaturated acids were selectively esterified at thesn-2-position. The triglyceride carbon number distribution of milk from the above species (with the exception of the Weddell seal) was determined by gas liquid chromatography and compared to that predicted by the 1-random-2-random-3-random fatty acid distribution hypothesis. Agreement was excellent between observed and predicted composition for echidna, koala, Tammar wallaby, guinea pig and pig milk, and agreement was reasonable for dog, cat, horse and cow milk. Results are discussed in relation to biochemical mechanisms.     

Response of docosapentaenoic acids of rat heart phospholipids to dietary fat

Groups of male Holtzman strain rats were fed from weanling one of the following diets: 20% hydrogenated soybean fat (20% HF), and 20% HF plus 2%, 3% and 4% corn oil, respectively, for 20 weeks. The animals were killed, and the heart phospholipid fractions isolated by chromatographic procedures. The levels and distribution of the docosapolyenoic acids, especially 22∶5ω3, were compared among the animals fed the corn oil supplemented and nonsupplemented diets. Although dietary linolenate (18∶3ω3) level was very low in the nonsupplemented diet, 22∶5ω3 accounted for 8.4% of the total fatty acids of heart total phospholipids when this diet was fed-half the level of total eicosatetraenoic acids. The amounts of 22∶5ω3 were decreased by corn oil supplementation of the diet and got down to the “normal” range of 2.0–2.5% at corn oil supplementation levels greater than 2%. The docosapolyenoic acids were confined largely to the phosphatidylcholine and phosphatidylethanolamine classes of phospholipids. These findings are discussed from the standpoint of the structural role of the phospholipids in the heart subcellular fractions.     

Positional distribution of the fatty acids in the triglycerides of mango (Mangifera indica) kernel fat

Triglycerides of mango seed kernel fat contain, depending on the variety, 32.4–44.0% of stearic acid and 43.7–54.5% of oleic acid. Palmitic and linoleic acids represent, respectively, 5.9–9.1% and 3.6–6.7% of the fatty acids. The triglycerides also contain minor amounts of arachidic and linolenic acids. Palmitic, stearic and arachidic acids were almost exclusively distributed among thesn-1-andsn-3-positions. Oleic acid represented 85–89% of the fatty acids at thesn-2-position. Oleic acid at thesn-1- andsn-3-positions showed a preference for thesn-1-position. Linoleic acid was mainly esterified at thesn-2-position. The amounts of saturated fatty acids, i.e., palmitic and stearic acids, and of oleic acid, at thesn-1- and sn-3-positions, were linearly related to their respective contents in the total triglycerides.     

The effect of hibernation on the positional distribution of ethanolamineglycerophospholipid fatty acids in hamster brain membranes

Microsomal and myelin membrane fractions were prepared from the brains of warm-adapted (room temperature) and hibernating Syrian Hamsters (Mesocricetus auratus). Ethanolamineglycerophospholipids were isolated and subjected to a fraction scheme to separate the fatty acids of the plasmenylethanolamine and the phosphatidylethanolamine 1 and 2 positions. The major changes in microsomal phosphatidylethanolamine with hibernation were relative increases in 18∶1 at the 1 position and 20∶4(n−6) in the 2 position. In myelin, 18∶1 increased and 18∶0 decreased at the 1 position while the 2 position showed an increase in 16∶0 and a decrease in 22∶6(n−3). Plasmenylethanolamine fatty acids also changed in microsomes and myelin although the magnitudes were not as great and confined to longer chain fatty acids. In both membranes, fatty acid alterations were position-specific, and no new types of fatty acids were introduced at any position.     

A comparison of acyltransferase activities in vitro with the distribution of fatty acids in lecithins and triglycerides in vivo

The location and configuration of a double bond in a fatty acid influences the rate of its acyltransferase-catalyzed esterification to form lecithin and its distribution in vivo between the primary and secondary positions of triglycerides and lecithins. Saturated acids of shorter chain length are transferred at rates similar to the long chain unsaturated acids. The positional distributions of acids in the diglyceride units of liver triglycerides appear to be similar to that found in the lecithins. Acyltransferase activities measured in vitro have a considerable predictive value in terms of the ultimate distribution of fatty acids in glycerolipids in vivo.     

Specific distribution of fatty acids in the triglycerides of rainbow trout adipose tissue. Influence of temperature

In the trout, the unsaturated fatty acids are preferentially located in the β-position and the saturated fatty acids in the α-position of triglycerides. This fatty acid distribution is retained even with diets containing lard. The fish are, therefore, able to modify completely the fatty acid distribution of dietary triglycerides. There is no retention of the β-monoglyceride structure during the biosynthetic processes. However, the modification of the dietary fatty acid distribution by the trout seems to be more difficult at 18 C than at 10C. Presented in part at the 13th World Congress of the International Society for Fat Research, Marseille, France, 1976.     

Surface tension of fatty acids and triglycerides

Surface tension as a function of temperature was measured for four fatty acids (lauric, myristic, palmitic, and oleic) and two triglycerides (tricaprylin and tripalmitin). These surface tension measurements were performed using a K12 vers. 3.1 (Krüss GmbH) tensiometer at temperatures from 20 (or the melting point of each substance) to 90°C. The constants for a van der Waals-type correlation as well as for a linear equation are presented. Both equations are quite accurate, presenting mean deviations not exceeding 0.570%. Such correlation constants are valuable in the design or evaluation of processing equipment, especially that involving gas-liquid contact such as distillation and stripping columns, deodorizers, reactors, and equipment for physical refining.