Relationship of Cellular Fatty Acid Composition to Survival of Lactobacillus bulgaricus in Liquid Nitrogen

Concentrated cultures of Lactobacillus bulgaricus were prepared by resuspending cells grown in semisynthetic media in sterile 10% non-fat milk solids. The concentrated cultures were frozen in liquid nitrogen for 24 h. The cell suspensions exhibited decreased viability after storage, and the amount of death varied among the different strains tested. Storage stability of all strains examined was improved by supplementing the growth medium with sodium oleate. Radioisotopes were used to study the fate of sodium oleate with L. bulgaricus NCS1. [1-(14)C]sodium oleate was incorporated solely into the lipid portion of the cells, including both neutral and polar lipids. The fatty acid composition of L. bulgaricus NCS1, NCS2, NCS3, and NCS4 grown with and without sodium oleate was studied. The major fatty acids of strains NCS1, NCS2, and NCS3 grown without sodium oleate were dodecanoic, tetradecanoic, hexadecanoic, hexadecenoic, and octadecenoic acids. In addition to these, strain NCS4 contained C(19) cyclopropane fatty acid. The major fatty acids of all strains grown with sodium oleate were tetradecanoic, hexadecanoic, hexadecenoic, octadecenoic, and C(19) cyclopropane fatty acids. All strains grown in broth containing sodium oleate contained larger amounts of octadecenoic and C(19) cyclopropane fatty acid, and less saturated fatty acids than when grown without sodium oleate. Statistical analyses indicated that C(19) cyclopropane fatty acid was most closely related to stability of the lactobacilli in liquid nitrogen. A negative regression line that was significant at P < 0.001 was obtained when the cellular content of this fatty acid was plotted against death.     

Replacement of the aliphatic chains of Clostridium acetobutylicum by exogenous fatty acids: regulation of phospholipid and glycolipid composition.

The membrane lipid aliphatic chains of Clostridium acetobutylicum ATCC 4259 have been extensively modified by growth in biotin-free medium containing vitamin-free casein hydrolysate supplemented with either elaidic acid, oleic acid, or mixtures of palmitic and oleic acids. Growth with elaidic acid resulted in polar lipids containing 88.6% 18:1 acyl chains and 94.5% 18:1 ether-linked chains. Growth with oleic acid resulted in comparable levels of enrichment of the lipids with 18:1 chains and C19 chains containing cyclopropane rings. When cells were grown with mixtures of palmitic and oleic acids, the ether-linked chains of the plasmalogens were greater than or equal to 64% 18:1 plus C19 chains containing cyclopropane rings at all ratios of oleic to palmitic acid in the medium. The acyl chains reflected the palmitic acid content of the medium more closely. Marked changes were observed in both phospholipid and glycosyldiglyceride compositions as the lipid acyl and ether-linked chains became more enriched with unsaturated and cyclopropane chains. The ratio of the glycerol acetal of plasmenylethanolamine to phosphatidylethanolamine increased, the ratio of cardiolipin to phosphatidylglycerol decreased, and the ratio of diglycosyldiglyceride to monoglycosyldiglyceride increased. However, the monoglycosyldiglyceride/diglycosyldiglyceride ratio was lower for cells grown on 100% oleic acid than for cells grown on 60 or 80% oleic acid. In the membranes of cells grown on 100% oleic acid, the ratio of glycolipids to phospholipids was lower than that found in cells grown on 60% oleic acid. These results indicate that C. acetobutylicum regulates its polar lipid composition in a complex manner involving phospholipids and glycosyldiglycerides. These changes can affect the equilibria between those lipids that form bilayers and those lipids that tend to form nonlamellar phases when enriched with unsaturated aliphatic chains. Phosphoglycolipids of unknown structure were also observed in cells grown either with biotin or with fatty acids. The content of the most abundant phosphoglycolipid also varied with the degree of unsaturation of the cellular lipids.     

Some factors affecting cyclopropane acid formation in Escherichia coli

1. The fatty acid composition of the extractable lipids of Escherichia coli varied with growth conditions. 2. The principal fatty acids were palmitic acid, hexadecenoic acid, octadecenoic acid and the cyclopropane acids, methylenehexadecanoic acid and methyleneoctadecanoic acid. 3. Cyclopropane acid formation from monoenoic acids was increased by acid media, poor oxygen supply, or high growth temperature. 4. Cyclopropane acid formation was decreased by alkaline media, well oxygenated conditions, the presence of citrate, or lack of Mg(2+).     

Evidence for the Existence of an Aerobic Pathway for Synthesis of Monounsaturated Fatty Acids by Alcaligenes faecalis

Studies on the composition of total fatty acids of Alcaligenes faecalis harvested at different growth phases have been carried out. Ability of the organism to desaturate palmitic and stearic acid has also been tested. The organism contained palmitic (16:0), stearic (18:0), palmitoleic (16:1), cis-vaccenic (18:1), cyclopropane (17: big dn tri, open and 19: big dn tri, open), and three hydroxy acids. Increase in cyclopropane acids and corresponding decrease in monounsaturated acids in direct proportion to the age of the culture were observed, whereas other fatty acids remained relatively unaltered. A growing culture of the organism was found to desaturate [1-(14)C]palmitic acid supplied in the medium to hexadecanoic acid. Resting cells desaturated [1-(14)C]palmitic and [1-(14)C]stearic acid giving rise to about 50% of (14)C in the COOH group of corresponding monounsaturated fatty acids.     

The effects of temperature on the composition and physical properties of the lipids of Pseudomonas fluorescens

1. Pseudomonas fluorescens was grown at various temperatures between 5 degrees C and 33 degrees C. The extractable lipids from organisms at various stages of growth and grown at different temperatures were examined. 2. The extractable lipids contained phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine, and an ornithine-containing lipid. The relative amounts of these lipids did not vary significantly during growth or with the changes in growth temperature. 3. The major fatty acids were hexadecanoic, hexadecenoic and octadecenoic acids and the cyclopropane acids methylene-hexadecanoic and methylene-octadecanoic acids. The relative amount of unsaturated acids (including cyclopropane acids) did not change significantly during growth, but increased with decreasing temperature. 4. Phosphatidylethanolamines with different degrees of unsaturation and containing different amounts of cyclopropane acids were isolated from organisms grown at 5 degrees C and 22 degrees C and their surface and phase behaviour in water was investigated. Thermodynamic parameters for fusion and monolayer results for cyclopropane and other fatty acids were examined. 5. The surface pressure-area isotherms of phosphatidylethanolamines containing different amounts of unsaturated fatty acids show small differences but the individual isotherms remain essentially unchanged over the temperature range 5-22 degrees C. X-ray-diffraction methods show that the structures (lamellar+hexagonal) formed in water by phosphatidylethanolamine, isolated from organisms grown at 5 degrees C and 22 degrees C, are identical when compared at the respective growth temperatures. This points to a control mechanism of the physical state of the lipids that is sensitive to the operating temperature of the organism. 6. The molecular packing of cyclopropane acids is intermediate between that of the corresponding cis- and trans-monoenoic acids. However, substitution of a cyclopropane acid for a cis-unsaturated acid has insignificant effects on the molecular packing of phospholipids containing these acids.     

Degradation of bacterial cyclopropane acids with boron trihalide reagents

The cellular fatty acid compositions of Pseudomonas diminuta UC 501 and Streptococcus mutans OMZ-61 were compared in samples processed by a saponification-methylation procedure (method S) and in samples processed by a transesterification procedure (method T). All samples were heated in an inert atmosphere of nitrogen. The major acids found in samples of P. diminuta treated by method S included 16:0 (palmitic), 18:1 (octadecenoic) and 19 cyc (19 carbon cyclopropane acid). Those found in samples of S. mutans treated by the same method included 16:0, 18:1. 18:0 (stearic), 20:1 (eicosenoic), 20:0 (eicosanoic), 19 cyc and 21 cyc. When method T was used to process samples of both cultures, the cyc acids were degraded and artifacts were produced. Transesterification with boron trihalide reagents is therefore not recommended for routine analysis of bacterial fatty acids.     

Cellular Lipid and Fatty Acid Compositions of Burkholderia pseudomallei Strains Isolated from Human and Environment in Viet Nam

Viet nam is known as an endemic area of melioidosis but its etiologic agent originated in Viet nam was not extensively studied. For the first time, we analyzed the cellular lipid and fatty acid compositions of 15 Vietnamese isolates of Burkholderia pseudomallei, 10 from humans and 5 from the environment. Cellular lipid compositions were analyzed by two-dimensional thin-layer chromatography on silica gel G plates. Cellular fatty acid methyl esters were analyzed by gas chromatography (GC) and gas chromatography/mass spectrometry (GC/MS). The major lipids in all the isolates were phosphatidylglycerol (PG), two forms of phosphatidylethanolamine (PE-1 and PE-2), and two forms of ornithine-containing lipid (OL-1 and OL-2). PE-1 contained non-hydroxy fatty acids at both sn-1 and ?2 positions, while PE-2 possessed 2-hydroxy fatty acids and non-hydroxy fatty acids in a ratio of 1: 1. Since snake venom phospholipase A₂ digestion of PE-2 liberated 2-hydroxy fatty acids, it was confirmed that these acids are at the sn-2 position of glycerol moiety. In both OL-1 and OL-2, amide-linked fatty acid was 3-hydroxy palmitic acid (3-OH-C16: 0), while ester-linked fatty acids were non-hydroxy acids in OL-1 and 2-hydroxy acids in OL-2. The total cellular fatty acid compositions of the test strains were characterized by the presence of 2-hydroxy palmitic (2-OH-C16: 0), 2-hydroxy hexadecenoic (2-OH-C16: 1), 2-hydroxy octadecenoic (2-OH-C18: 1), 2-hydroxy methylene octadecanoic (2-OH-C19CPA), 3-hydroxy myristic (3-OH-C14: 0) and 3-hydroxy palmitic (3-OH-C16: 0) acids. There were significant differences in the concentration of hexadecenoic (C16: 1), methylene hexadecanoic (C17CPA), octadecenoic (C18: 1) and methylene octadecanoic (C19CPA) acids among the Vietnamese isolates of B. pseudomallei. However, no significant difference was observed in cellular lipid and fatty acid components between strains of human and environmental origins.     

Adaptational changes of fatty acid composition and the physical state of membrane lipids following the change of growth temperature in Yersinia enterocolitica.

Yersinia enterocolitica is capable of growing in a broad range of temperatures from 4 to 45 C. How this organism alters its membrane lipids in response to the change of growth temperature is very interesting. The fatty acids of membrane lipids of cells cultured at 5, 15, 25 and 37 C were analyzed and the physical states of these membrane lipids were characterized. The major phospholipids of this bacterium were phosphatidylethanolamine, phosphatidylglycerol, cardiolipin, lysophosphatidylglycerol and lysophosphatidylethanolamine. No significant difference in phospholipid composition in response to culture temperatures was observed. It was reported in our previous paper that the major fatty acids of membrane phospholipids of Y. enterocolitica were C15:0, C16:0, C16:1, cyclopropane C17:0 and C18:0. Some differences in the fatty acid composition were, however, observed with the change of culture temperature. When the culture temperature was raised, the saturated and cyclopropane fatty acids substantially increased and the unsaturated ones decreased. A reverse phenomenon was observed when culture temperature was lowered. From the viewpoints of membrane physical state, adaptational changes were analyzed using a nylon microcapsule method. Phase transition in membrane lipids of cells grown at each culture temperature took place in the range of about 5 C below and about 10 C above the culture temperature. It is, therefore, considered that Y. enterocolitica maintains its membrane rigidity and fluidity in response to growth temperature by changing the membrane fatty acid composition.     

EFFECT OF TEMPERATURE ON THE COMPOSITION OF FATTY ACIDS IN ESCHERICHIA COLI.

Marr, Allen G. (University of California, Davis) and John L. Ingraham. Effect of temperature on composition of fatty acids in Escherichia coli. J. Bacteriol. 84:1260-1267. 1962.-Variations in the temperature of growth and in the composition of the medium alter the proportions of individual fatty acids in the lipids of Escherichia coli. As the temperature of growth is lowered, the proportion of unsaturated fatty acids (hexadecenoic and octadecenoic acids) increases. The increase in content of unsaturated acids with a decrease in temperature of growth occurs in both minimal and complex media. Cells harvested in the stationary phase contained large amounts of cyclopropane fatty acids (methylenehexadecanoic and methylene octadecanoic acids) in comparison with cells harvested during exponential growth. Cells grown in a chemostat, limited by the concentration of ammonium salts, show a much higher content of saturated fatty acids (principally palmitic acid) than do cells harvested from an exponentially-growing batch culture in the same medium. Cells grown in a chemostat, limited by the concentration of glucose, show a slightly higher content of unsaturated fatty acids than cells from the corresponding batch culture. The results do not indicate a direct relation between fatty acid composition and minimal growth temperature.     

Synthesis of fatty acids from [1-14C]acetylcoenzyme A in subcellular particles of rat epididymal adipose tissue

1. Mitochondrial and microsomal fractions of rat epididymal adipose tissue incorporated [1-(14)C]acetyl-CoA equally well into various fatty acids by a chain-elongation mechanism. C(18) and C(20) fatty acids were the two major products, and comprised about 80% of the total fatty acids synthesized in both particles. 2. When incubated in air, mitochondria synthesized stearic acid, octadecenoic acid and eicosamonoenoic acid in almost equal amounts (about 20% each), whereas in microsomal fractions, the synthesis of octadecenoic acid was more than fivefold the stearic acid formation. In both fractions, major components of synthesized monoenoic fatty acids were the Delta(11:12) isomers. Hexadecenoic acid and octadecenoic acid from whole adipose tissue contained approx. 11 and 14% of the Delta(11:12) isomer respectively. 3. When mitochondria or microsomal fractions were incubated in nitrogen, there was increased synthesis of stearic acid and palmitic acid and less of C(16) and C(18) monoenoic acids; synthesis of C(20) acids remained predominantly of the monoenoic acids. Determination of the position of the double bond in the monoenoic acids supported the view that the synthesis of hexadecenoic acid and octadecenoic acid involves a desaturase activity, whereas eicosamonoenoic acid and eicosadienoic acid are formed only by elongation of endogenous fatty acids. 4. Most of the radioactivity was found in free fatty acids (63%) and the phospholipid (26%) fraction. In phospholipids, phosphatidylcholine and phosphatidylethanolamine were the two major components. 5. Most of the fatty acids synthesized, including those not normally found in particle lipids (arachidic acid, eicosamonoenoic acid and eicosadienoic acid) were distributed fairly evenly in the phospholipid and free fatty acid fractions. However, stearic acid was found predominantly in the phospholipid fraction.     

Hyperthermic sensitivity and growth stage in Escherichia coli

Hyperthermic sensitivities of Escherichia coli B/r and Bs-1 were determined for lag-, midlog-, and stationary-phase cells at 47, 48, and 49 degrees C. In both strains midlog-phase cells were strikingly more heat sensitive (100-fold greater killing after 4 h at 48 degrees C) than stationary-phase cells, with intermediate sensitivity for lag-phase cells. In contrast to the reported difference in the radiation sensitivity between these two strains, very little difference in heat sensitivity was seen. Patterns of fatty acid composition of both strains were very similar at each phase of growth. From midlog to stationary phase, 16:1 and 18:1 unsaturated fatty acids decrease from 16 and 30% to 0.5 and 3%, respectively, while the C17 and C19 cyclopropane fatty acids increase from 7 and 3% to 22 and 25%, respectively. Concomitant with these changes in fatty acid composition, substantially higher membrane microviscosity values were recorded for stationary-phase cells. Total membrane microviscosity was positively associated with the C17 and C19 cyclopropane fatty acid composition and with cell survival following hyperthermia. In contrast to hyperthermic sensitivity, radiation survival differences between B/r and Bs-1 are little affected by growth stage. We propose that these results are consistent with a critical influence of membrane lipids on cellular hyperthermic sensitivity and further that the target sites for radiation and hyperthermia are different in these cells.     

Effects of temperature and nutritional changes on the fatty acids of agmenellum quadruplicatum.

The fatty acid composition of the blue-green bacterium Agmenellum quadruplicatum was examined under a wide variety of growth conditions. The fatty acid composition was found to undergo significant changes with variations in temperature, media composition, and growth phase (log versus stationary). With increasing growth temperature (20 to 43 C) log-phase cells exhibited an increase in saturated fatty acids (38.4% at 20 C to 63.6% at 43 C). Striking changes were seen with some of the individual fatty acids such as 18.3, which made up 16.0% of the total fatty acid at 20 C but was not neasurable at 43 C. Fatty acid 12:0 was not measurable at 20 C but made up 16.3% of the total fatty acids at 43 C. Cell lipids were separated into neutral lipid, glycolipid, and very polar liquid fractions. The neutral lipid fraction was composed almost entirely of 12 carbon fatty acids (12:0, 12:1). Glycolipid and very polar lipids were more similar in their fatty acid composition when compared to the total cellular fatty acids, although they did lack 12 carbon fatty acids. The total of 12 carbon fatty acids in the cell can be used as an indicator of the amount of neutral lipid present.     

Temperature- and growth-phase-regulated changes in lipid fatty acid structures of psychrotolerant groundwater Proteobacteria

Cellular fatty acid compositions of five psychrotolerant groundwater isolates representing alpha- and beta-Proteobacteria were studied at temperatures ranging from 8 to 25 degrees C. Unsaturation of straight-chain fatty acids was the most common response to decreasing temperature and was detected in four of the isolates. On solid media, decrease of temperature resulted in a decrease of cyclopropane fatty acids in beta-proteobacterial isolates. The formation of cyclopropane fatty acids depended, however, to a greater extent on the growth phase than the temperature and increased drastically as the cells entered stationary phase. The alpha-proteobacterial isolates contained a branched C(19:1) fatty acid. The formation of the branched C(19:1) increased during growth in the same way as the cyclopropane fatty acids in beta-proteobacterial strains, indicating possibly an analogous formation of the branched fatty acid by methylation of the 18:1 fatty acid. Sphingomonas sp. K6 possessed a novel temperature-induced modification of lipid fatty acids. As temperature decreased from 25 to 8 degrees C, the fatty acid composition shifted from predominantly even-carbon fatty acids to odd-carbon fatty acids. The results show completely different fatty acid modifications in two strains of the same genus Sphingomonas.     

The glycolipids of Lactobacillus casei A.T.C.C. 7469

1. The lipids were extracted from Lactobacillus casei A.T.C.C. 7469 with chloroform-methanol mixtures. The glycolipids were obtained by chromatography on silicic acid and DEAE-cellulose (acetate form). 2. Hydrolysis of the glycolipids with alkali gave two glycerol glycosides and a mixture of fatty acids. 3. The glycosides were separated and their structures elucidated. The major component was O-alpha-d-galactopyranosyl-(1-->2)-O-alpha-d-glucopyranosyl-(1-->1)-glycerol and the minor component O-alpha-d-glucopyranosyl-(1-->6)-O-alpha-d-galactopyranosyl-(1-->2)-O-alpha-d-glucopyranosyl-(1-->1)-glycerol. 4. Analysis of the fatty acids by gas-liquid chromatography showed that they were predominantly palmitic acid, octadecenoic acid and lactobacillic acid.     

Fatty acid composition of selected prosthecate bacteria

The cellular fatty acid composition of 14 strains of Caulobacter species and types, two species of Prosthecomicrobium, and two species of Asticcacaulis was determined by gas-liquid chromatography. In most of these bacteria, the major fatty acids were octadecenoic acid (C18:1), hexadecenoic acid (C16:1) and hexadecanoic acid (C16:0). Some cyclopropane and branched chain fatty acids were detected in addition to the straight chained acids. Hydroxytetradecanoic acid was an important component of P. enhydrum but significant amounts of hydroxy acids were not detected in other prosthecate bacteria examined.Abbreviations DEGS diethylene glycol succinate - A measare of association Dedicated to Prof. R. Y. Stanier on the occasion of his 60th birthday     

Trans fatty acids promote the growth of some Lactobacillus strains

Five Lactobacillus strains (2 L. gasseri, 2 L. plantarum and 1 L. reuteri) were cultured in modified MRS medium containing fatty acids (FAs) instead of Tween 80 for 24 h at 37 degrees C, to learn the effect of saturated and unsaturated FAs on the Lactobacillus growth. Free FAs included palmitic (16:0), palmitoleic (c9-16:1), stearic (18:0), oleic (c9-18:1), elaidic (t9-18:1), cis-vaccenic (c11-18:1), vaccenic (t11-18:1), linoleic (c9, c12-18:2), conjugated linoleic (c9, t11- and t10, c12-18:2), alpha-linolenic (c9, c12, c15-18:3), alpha-eleostearic (c9, t11, t13-18:3), eicosapentaenoic (20:5), and docosahexaenoic (22:6) acids. Among free FAs, oleic acid stimulated the growth of all Lactobacillus strains, whereas palmitoleic acid had almost no affect on the Lactobacillus growth. Saturated FAs such as stearic and palmitic acids inhibited or did not affect the Lactobacillus growth. Polyunsaturated FAs such as alpha-linolenic, eicosapentaenoic and docosahexaenoic acids strongly inhibited the Lactobacillus growth at 7.6 x 10(-4) m. Octadecenoic acids such as oleic, elaidic, cis-vaccenic and vaccenic acids remarkably promoted the growth of L. gasseri, regardless of the different double bond positions and configurations. When oleic or cis-vaccenic acid was incubated with L. gasseri, the FAs was transformed to cyclopropane FAs (methyleneoctadecanoic acids) after incorporation into the cells. On the other hand, trans FAs such as elaidic and vaccenic acids incorporated into the cells were not converted to another FAs. Conjugated linoleic and alpha-eleostearic acids having a trans double bond promoted the Lactobacillus growth. The growth of L. gasseri was also stimulated by trans-rich free FAs from hydrogenated canola and fish oils. These results showed that octadecenoic acid and trans FAs had strong promotion activities for the Lactobacillus growth due to their incorporation into membrane lipids.     

The effect of fluoride on the growth and fatty acid composition of Sphagnum fimbriatum at two temperatures

The fatly acid composition of different lipid fractions (neutral, glyco- and phospholipids) was studied in Sphagnum fimbriatum Wils, gametophytes grown in aseptic cultures at two temperatures (15°C and 25°C). The effect of a growth-retarding concentration (0.1 mM) of KF was also investigated. Fifteen-day treatment with KF affected the fatty acid composition more strongly at the higher than at the lower temperature. The proportion of palmitic acid (16:0) increased but the proportion of linoleic (18:2) decreased in all the lipid fractions, and that of linolenic (18:3) acid decreased in the fractions containing glyco- and neutral lipids. This indicates that the fluoride ions inhibit lengthening of the fatty acid chain. Compared with gametophytes grown at 25°C, material cultivated at 15°C had a much higher proportion of a highly unsaturated fatty acid, linolenic acid (18:3), in all the lipid fractions, but a lower proportion of oleic acid (18:1) in the neutral and phospholipids, and a lower proportion of linoleic (18:2) acid in all three fractions.     

Effect of copper on membrane lipids and on methane monooxygenase activity of Methylococcus capsulatus (Bath)

The effect of copper supplementation on growth, methane monooxygenase activity and lipid composition of Methylococcus capsulatus (Bath) was studied. Copper increased biomass yield, methane monooxygenase activity and phospholipid content from 7.7 to 10.2% of dry weight. Cells from copper-deficient and copper supplemented cultures contained the same major fatty acids but in the presence of copper only the contents of C16:0 and the three C16:1 isomers were increased while the contents of C14:0 and cyclic C17:0 remained unchanged. Phosphatidylethanolamine (PE), phosphatidylcholine (PC), phosphatidylglycerol and cardiolipin were analysed amongst the polar lipids. PE was the main component (about 60 mol-%) but the most notable copper-induced increment occurred in the proportion of PC, from about 10 to 16 mol-%. Concomitantly with this increment the fatty acids of PC were changed so that the mol-% of C16: 1 isomers were increased at the expense of other acids. Similar trends were seen also in the fatty acid compositions of other polar lipid fractions. It is therefore concluded that phosphatidylcholine would be one of the key factors when the role of membranous lipids in methane monooxygenase activity is to be considered.     

Synthesis of lipids from acetate by human preputial and abdominal skin in vitro

Lipogenesis in vitro from acetate-1-(14)C was studied in human preputial skin and abdominal skin. Radioactive lipids were separated by column chromatography on Florisil and by thin-layer chromatography on silica gel. Radioactivity was incorporated chiefly into the triglyceride, sterol, and polar lipid fractions, while lesser amounts of (14)C were found in the hydrocarbon, wax, diglyceride, monoglyceride, and fatty acid fractions; labeling of steryl esters was minimal. On thin-layer chromatography, the radioactive polar lipids had mobilities similar to lysolecithin, phosphatidyl choline, phosphatidyl ethanolamine, and phosphatidic acid. The radioactive fatty acids of the different lipid fractions were separated by gas-liquid chromatography. The major (14)C-labeled acids were 16:0 and 18:0. Radioactivity was also detected in acids 14:0, 15:0, 16:1, 18:1, 18:2, 20:0, 20:1, 22:0, 24:0, 24:1, and 26:0. No radioactivity could be detected in arachidonic acid, although this fatty acid comprises 9% of the chromatographed fatty acids. The pattern of incorporated (14)C was different from the percentage mass composition of the fatty acids. Skin is therefore active in the biosynthesis of a wider variety of lipids than previously demonstrated.     

The lipid composition of a halotolerant species of Staphylococcus epidermidis.

Studies were carried out on the lipid composition of a halotolerant Staphylococcus epidermidis isolated in pure culture from a growth medium for extreme halophiles containing 25% NaCl. The four major polar lipid components in this bacterium were found to be: (a) glycerophosphoryl diglucosyl diglyceride (10% by weight) with structure 3(1)-O-(-sn-glycerol-1-phosphoryl-6'-O=(beta-D glucopyranosyl-(1 leads to 6)- O-beta-D-glucopyranosyl)-1(3),2-diacyl-sn-glycerol; (b) diglucosyl diglyceride (15% by weight) with structure 3(1)-O-(beta-D-glucopyranosyl (1 leads to 6)-O-beta-D-glucopyranosyl)-1(3),2-diacyl-sn-glycerol; (c) monoglucosyl diglyceride (3% by weight) with structure 3(1)-O-(beta-D-glucopyranosyl)-1(3),2-diacyl-sn-glycerol, and (d) phosphatidylglycerol (60% by weight) with structure 1,2 diacyl-sn-glycero-3-phosphoryl-1'-sn-glycerol. Phosphatidic acid, cardiolipin, lysophosphatidylglycerol and three unidentified phospholipids were also detected in small amounts. Each lipid component had essentially the same fatty acid composition namely, anteiso-15:0 (60-75%), anteiso-17:0 (18-24%), iso-17:0 (8--10%), and small amounts of palmitic and stearic acids (2-5%). The fatty acids were non-randomly distributed in phosphatidylglycerol, the shorter chain anteiso 15:0 fatty acid being exclusively esterified to the 2-position and the longer chain anteiso- and iso-17:0 fatty acids at the 1-position. The fatty acid composition was not affected by increaseing NaCl content in the medium in the rande 0--15% but the proportion of anteiso-15:0 increased greatly when the salt concentration was increased to 25%. The proportions of ionic polar lipids were modified to give an increased net negative charge per mol ionic lipids when NaCl in the medium was increased from 15 to 25%, but the proportions of neutral glycolipids remained fairly constant.