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Temperature and nutrient availability control growth rate and fatty acid composition of facultatively psychrophilic Cobetia marina strain L-2.

A facultative psychrophilic bacterium, strain L-2, that grows at 0 and 5 degrees C as minimum growth temperatures in complex and defined media, respectively, was isolated. On the basis of taxonomic studies, strain L-2 was identified as Cobetia marina. The adaptability of strain L-2 to cold temperature was higher than that of the type strain and of other reported strains of the same species. When the bacterium was grown at 5-15 degrees C in a defined medium, it produced a high amount of trans-unsaturated fatty acids. By contrast, in a complex medium in the same temperature range it produced a low amount of trans-unsaturated fatty acids. In the complex medium at 5 degrees C, the bacterium exhibited a three-fold higher growth rate than that obtained in the defined medium. Following a temperature shift from 11 to 5 degrees C, strain L-2 grew better in complex than in defined medium. Furthermore, when the growth temperature was shifted from 0 to 5 degrees C both the growth rate and the yield of strain L-2 growing in complex medium was markedly enhanced. These phenomena suggest that an upshift of the growth temperature had a positive effect on metabolism. The effects of adding complex medium components to the defined medium on bacterial growth rate and fatty acid composition at 5 degrees C were also studied. The addition of yeast extract followed by peptone was effective in promoting rapid growth, while glutamate addition was less effective, resulting in a cis-unsaturated fatty acid ratio similar to that of cells grown in the complex medium. These results suggest that the rapid growth of strain L-2 at low temperatures requires a high content of various amino acids rather than the presence of a high ratio of cis-unsaturated fatty acids in the cell membrane.

Cell Membrane↗

Characteristics of the myocardial PM-FABP: effect of diabetes mellitus.

Properties of the myocardial PM-FABP were studied in normal and STZ-diabetic rats. The fluorescent fatty acids trans-parinaric and cis-parinaric acids were used as analogs of straight-chain (saturated) and kinked-chain (unsaturated) fatty acids respectively. Parinaric acid binding was sensitive to trypsin. Trans-parinaric acid binding was more sensitive to this protease than the binding of cis-parinaric acid. Based on the difference in sensitivity of parinaric acid binding we believe that there are two separate binding sites associated with myocardial PM-FABP; one for unsaturated fats and the other for saturated fats. Diabetes enhanced both cis- and trans-parinaric acid binding capacity in cardiomyocytes; cis-parinaric acid by 2 fold and trans-parinaric acid by 2.6 fold. In addition, there was a concomitant accumulation of free fatty acids and triglycerides in the hearts of the diabetic animals. There was a 2.2 fold increase for fatty acids and a 1.6 fold increase for trigylcerides. This association between myocardial fatty acid build-up and enhanced myocardial PM-FABP during diabetes suggest that this carrier protein might have contributed to lipid accumulation in the hearts of the diabetic rats.

Animals↗

Pseudomonas putida NCTC 10936 balances membrane fluidity in response to physical and chemical stress by changing the saturation degree and the trans/cis ratio of fatty acids.

This study explored the capability of Pseudomonas putida NCTC 10936 to maintain homeoviscosity after changing the growth temperature, incubating resting cells at different temperatures or at a constant temperature in the presence of 4-chlorophenol (4-CP). After raising the growth temperature from 20 to either 30 or 35 degrees C, the degree of saturation of the organism's fatty acids increased and the ratio of trans to cis unsaturated fatty acids decreased somewhat. In contrast, after the incubation temperature of resting cells was raised (grown at 30 degrees C) from 20 to 30 or 35 degrees C the degree of saturation of the fatty acids remained nearly constant, while the ratio of trans to cis unsaturated fatty acids increased. Incubating resting cells (grown at 30 degrees C) at 20 degrees C in the presence of 4-CP again caused no major changes in the degree of saturation, but cis to trans conversion of unsaturated fatty acids was induced, with a corresponding increase in the trans/cis ratios. Increases in both the saturation degree of the fatty acids and the trans/cis ratio of the unsaturated fatty acids correlated with increases in the fluorescence anisotropy of 1,6-diphenyl-1,3,5-hexatriene intercalated in the bilayers of liposomes prepared from the cells of P. putida NCTC 10936. Electron transport phosphorylation (ETP) could be stabilized by adaptive adjustments in the fluidity of the cytoplasmic membrane mediated by changes in fatty acid composition such as those observed. Whether changes in the degree of saturation or in the trans/cis ratio are more effective can be decided by studying P. putida NCTC 10936.

Anisotropy↗

Use of an unsaturated fatty acid auxotroph of Saccharomyces cerevisiae to modify the lipid composition and function of mitochondrial membranes.

KD115 (ol1), an unsaturated fatty acid auxotroph of S. cerevisiae, was grown in a semi-synthetic medium supplemented with 3.3 x 10(-4) M palmitoleic (cis 16:1) or palmitelaidic (trans 16:1) acids. The parent strain S288C was studied as a control. The lipid composition (fatty acids, neutral lipids, and phospholipids), respiratory activity (O2 consumption), and ultrastructure were compared in mutant yeast grown with each unsaturated fatty acid supplement. The fatty acid supplement represented 70-80% of the yeast fatty acids. Yeast grown in trans 16:1 contained more squalene, a higher ratio of phosphatidylethanolamine (PE) to phosphatidylcholine (PC), and had 10-20% of the respiratory activity compared to the same yeast grown in cis 16:1. The mitochondrial morphology of yeast in each growth supplement was notably different. The use of mixtures of cis and trans 16:1 in different proportions revealed that the PE/PC ratio, the squalene content, the respiratory defect, and the mitochondrial morphology were all similarly dependent on the fraction of trans 16:1 in the mixtures. As little as 10-20% of cis 16:1 in the mixture was sufficient to abrogate the physiological effects of trans 16:1 on each of the parameters noted above. The combined effects of high content of trans unsaturated fatty acid and the altered phospholipid composition seem to account for the decrease in lipid fluidity, the defective structure and function of the mitochondrial membrane.

Fatty Acids↗

Fatty acid modulation of K+ channels in taste receptor cells: gustatory cues for dietary fat.

In an attempt to determine the chemosensory cues, if any, provided by fats in the oral cavity, we have performed patch-clamp recordings on isolated rat taste receptor cells during application of free fatty acids. Cis-polyunsaturated fatty acids, when applied extracellularly, inhibit delayed-rectifying K+ channels. In a subset of cells, these fatty acids also enhance inwardly rectifying K+ currents. Saturated, monounsaturated, and trans-polyunsaturated fatty acids have no significant effect on K+ currents. These effects do not involve activation of G protein-mediated pathways, including protein kinase C and protein kinase A, lipoxygenase pathways, cyclooxygenase pathways, or cytochrome P-450 pathways, consistent with direct effects on these ion channels or closely associated proteins. The net effect of fatty acids is to prolong stimulus-induced depolarizations of taste receptor cells, and we propose the effects on K+ channels represent the mechanism by which fats are detected by receptor cells in the oral cavity.

Animals↗

The cis/trans isomerization of the double bond of a fatty acid as a strategy for adaptation to changes in ambient temperature in the psychrophilic bacterium, Vibrio sp. strain ABE-1.

The major phospholipid, phosphatidylethanolamine (PE), of Vibrio sp. strain ABE-1 contains a unique trans-unsaturated fatty acid, 9-trans-hexadecenoic acid (16:1(9t], at the sn-2 position of the glycerol moiety. The major molecular species of PE that contain 16:1(9t) at the sn-2 position have either 9-cis-hexadecenoic acid (16:1(9c] or hexadecanoic acid (16:0) at the sn-1 position. The transition temperatures of the liquid-crystal to the gel phase of 16:1(9c)/16:1(9t)-PE and 16:0/16:1(9t)-PE were -3 degrees C and 38 degrees C, respectively, temperatures that were 31 degrees C and 18 degrees C higher than the corresponding temperatures for 16:1(9c)/16:1(9c)-PE and 16:0/16:1(9c)-PE. The proportion of 16:1(9c)/16:1(9t)-PE and 16:0/16:1(9t)-PE increased significantly in cells grown at 20 degrees C over that in cells grown at 5 degrees C. When cells grown at 5 degrees C were incubated at 20 degrees C in the presence of cerulenin, an inhibitor of the synthesis de novo of fatty acids, the level of 16:1(9t) increased almost in parallel with a concomitant decrease in the level of 16:1(9c) at the sn-2 position. These results suggest that 16:1(9c) is converted to 16:1(9t) by the cis/trans isomerization of the double bond in the fatty acid. This conversion is discussed as a possible strategy for adaptation by bacteria to changes in temperature.

Adaptation, Physiological↗

Effects of fats and fatty acids on blood lipids in humans: an overview.

Differences in dietary fatty acid structure induce marked differences in lipid and lipoprotein concentrations in plasma from fasting subjects. Under metabolic-ward conditions, replacement of carbohydrates by lauric, myristic, and palmitic acids raise both low-density-lipoprotein (LDL) and high-density-lipoprotein (HDL) cholesterol whereas stearic acid has little effect. Oleic and linoleic acids raise HDL and slightly lower LDL; all fatty acids lower fasting triglycerides when substituted for carbohydrates. Trans monounsaturates lower HDL and raise LDL and lipoprotein(a). The fatty acids in unhydrogenated fish oil potently lower triglycerides, with variable effects on LDL. Of the commercial fats, palm-kernel and coconut oil are the most hypercholesterolemic, followed by butter and palm oil. Replacement of hard fats rich in lauric, myristic, or palmitic acids or trans fatty acids by unsaturated oils will lower LDL, but replacement by carbohydrates will in addition decrease HDL and increase triglycerides. In free-living subjects, high-oil diets could lead to obesity, undoing the favorable effects on HDL and triglycerides.

Cholesterol↗

Effects of substrate, passage rate, and pH in continuous culture on flows of conjugated linoleic acid and trans C18:1.

A dual-flow continuous culture system consisting of 4 fermenters was used in a 4 x4 Latin square design. The objective of the research was to evaluate the effects of solid dilution rate (SDR), pH, and concentration of linoleic acid (LA) in the feed mixture on the production of conjugated linoleic acid (CLA) and trans-C18:1. The 4 treatments were 1) control = pH 6.5, 1% LA, 4%/h SDR; 2) high solid dilution rate (HSDR) = pH 6.5, 1% LA, 8%/h SDR; 3) high linoleic acid (HLA) = pH 6.5, 3% LA, 4%/h SDR; and 4) low pH (LPH) = pH 5.8, 1% LA, 4%/h SDR. Inoculum was collected 6 h after feeding from a cow fed 40% alfalfa hay and 60% grain. Liquid dilution rate was held at 0.12/h. All treatments except HLA contained 2% tallow. The LA was dissolved in buffer and continuously infused into the fermenters. The CLA flows were 16.5, 20.4, 23.2, and 25.2 mg/d for control, HSDR, HLA, and LPH, respectively. Compared with control, LPH increased flows of CLA, cis-C18:1, and C18:2, and decreased flow of C18:0. The neutral detergent fiber (NDF) and acid detergent fiber (ADF) digestibilities were not affected by pH. The HSDR tended to increase CLA flow compared to control, possibly because a shorter solid retention time led to incomplete biohydrogenation (BH). The NDF and ADF digestibilities and bacterial numbers were reduced by HSDR. With more LA available as a substrate for CLA, HLA resulted in a higher flow of CLA than control. The HLA resulted in the highest acid detergent fiber and fatty acid digestibilities, bacterial numbers, and BH. Increasing solids passage rate, reducing pH, and increasing dietary LA appears to increase in vitro CLA production.

Acetates↗

Differential biohydrogenation and isomerization of [U-(13)C]oleic and [1-(13)C]oleic acids by mixed ruminal microbes.

The additional mass associated with 13C in metabolic tracers may interfere with their metabolism. The comparative isomerization and biohydrogenation of oleic, [1-(13)C]oleic, and [U-13C]oleic acids by mixed ruminal microbes was used to evaluate this effect. The percent of stearic, cis-14 and -15, and trans-9 to -16 18:1 originating from oleic acid was decreased for [U-(13)C]oleic acid compared with [1-(13)C]oleic acid. Conversely, microbial utilization of [U-(13)C]oleic acid resulted in more of the 13C label in cis-9 18:1 compared with [1-(13)C]oleic acid (53.7 vs. 40.1%). The isomerization and biohydrogenation of oleic acid by ruminal microbes is affected by the mass of the labeled tracer.

Analysis of Variance↗

Direct activation of purified protein kinase C by unsaturated fatty acids (oleate and arachidonate) in the absence of phospholipids and Ca2+.

Unsaturated fatty acids (oleic acid and arachidonic acid) activate purified protein kinase C independently of phospholipid and Ca2+. Oleic acid activation of protein kinase C is as effective as phosphatidylserine and Ca2+. Ka values for oleic acid and arachidonic acid are 50 and 53 microM, respectively. In contrast to the cis fatty acids, a trans form (elaidic acid) or a saturated fatty acid (stearic acid) has little or no effect on protein kinase C activation. If cis fatty acid liberation is physiologically important, this suggests that another mechanism may exist for protein kinase C activation, in addition to phospholipase C/phosphatidylinositol turnover signaling, possibly via the liberation of cis fatty acids by the Ca2+-dependent phospholipase A2 system.

Animals↗

Replacement of partially hydrogenated soybean oil by palm oil in margarine without unfavorable effects on serum lipoproteins.

We have compared the effects of three different margarines, one based on palm oil (PALM-margarine), one based on partially hydrogenated soybean oil (TRANS-margarine) and one with a high content of polyunsaturated fatty acids (PUFA-margarine), on serum lipids in 27 young women. The main purpose of the study was to test if replacement of trans fatty acids in margarine by palmitic acid results in unfavorable effects on serum lipids. The sum of saturated fatty acids (12:0, 14:0, 16:0) was 36.3% of total fatty acids in the PALM-diet, the same as the sum of saturated (12:0, 14:0, 16:0) (12.5%) and trans (23.1%) fatty acids in the TRANS-diet. This sum was 20.7% in the PUFA-diet. The content of oleic acid was 37.9, 35.2, and 38.6%, respectively, in the three diets, whereas linoleic acid amounted to 16, 13.5, and 27.3%, respectively. Total fat provided 30-31% and the test margarines 26% of total energy in all three diets. The subjects consumed each of the diets for 17 d in a Latin-square crossover design. There were no significant differences in total cholesterol, low density lipoprotein (LDL)-cholesterol and apolipoprotein B (apoB) between the TRANS- and the PALM-diets. High density lipoprotein (HDL)-cholesterol and apoA-1 were significantly higher on the PALM-diet compared to the TRANS-diet whereas the ratio of LDL-cholesterol to HDL-cholesterol was lower, although not significantly (P = 0.077) on the PALM-diet. Total cholesterol, LDL-cholesterol, and apoB were significantly lower on the PUFA-diet compared to the two other diets. HDL-cholesterol was not different on the PALM- and the PUFA-diets but it was significantly lower on the TRANS-diet compared to the PUFA diet. Compared to the PUFA-diet the ratio of LDL- to HDL-cholesterol was higher on both the PALM- and the TRANS-diets whereas apoA-1 was not different. Triglycerides and lipoprotein (a) were not significantly different among the three diets. We concluded that nutritionally, palmitic acid from palm oil may be a reasonable alternative to trans fatty acids from partially hydrogenated soybean oil in margarine if the aim is to avoid trans fatty acids. A palm oil-based margarine is, however, less favorable than one based on a more polyunsaturated vegetable oil.

Adult↗

cis-4,7,10,trans-1 3-22:4 fatty acid distribution in phospholipids of pectinid species Aequipecten opercularis and Pecten maximus.

The distribution of cis-4,7,10,trans-13-docosatetraenoic (c4,7,10,t13-22:4), a peculiar FA previously isolated in the glycerophospholipids of some pectinid bivalves, was investigated in glycerophospholipid classes and subclasses of separated organs (gills, mantle, gonads, and muscle) of the queen scallop Aequipecten opercularis and the king scallop Pecten maximus. Plasmalogen (Pls) and diacyl + alkyl (Ptd) forms of serine, ethanolamine, and choline glycerophospholipids were isolated by HPLC and their FA compositions analyzed by GC-FID. PIs and Ptd forms of serine glycerophospholipids (PlsSer and PtdSer), and to a lesser extend the Pls form of ethanolamine glycerophospholipids (PlsEtn), were found to be specifically enriched with c4,7,10,t13-22:4. This specificity was found to decrease in the tested organs in the following order: gills, mantle, gonad, and muscle. In gills, c4,7,10,t13-22:4 was shown to be the main unsaturated FA of serine glycerophospholipids in both Pls and Ptd forms (23.8 and 19.4 mol%, respectively, for A. opercularis, and 21.0 and 26.2 mol% for P. maximus). These results represent the first comprehensive report on the FA composition of plasmalogen serine subclass isolated from pectinid bivalves. The specific association of the PlsSer with the c4,7,10,t13-22:4 for the two pectinid species can be paralleled to the specific association of the PlsSer with the non-methylene interrupted (NMI) FA and 20:1 (n-11) observed in mussels, clams, and oysters (Kraffe, E., Soudant, P., and Marty, Y. (2004) Fatty Acids of Serine, Ethanolamine and Choline Plasmalogens in Some Marine Bivalves, Lipids 39, 59-66.) This, led us to hypothesize a similar functional significance for c4,7,10,t13-22:4, NMI FA, and 20:1 (n-11) associated with PlsSer subclass of bivalves.

Animals↗

Modulation of voltage-dependent Ca channel current by arachidonic acid and other long-chain fatty acids in rabbit intestinal smooth muscle.

The effects of arachidonic acid (AA) and other long-chain fatty acids on voltage-dependent Ca channel current (ICa) were investigated, with the whole cell patch clamp method, in longitudinal smooth muscle cells of rabbit ileum. 10-30 microM AA caused a gradual depression of ICa. The inhibitory effect of AA was not prevented by indomethacin (10 microM) (an inhibitor of cyclooxygenase) or nordihydroguaiaretic acid (10 microM) (an inhibitor of lipoxygenase). 1-(5-Isoquinolinesulfonyl)-2-methylpiperazine (H7; 25-50 microM) or staurosporine (2 microM) (inhibitors of protein kinase C) did not block the AA-induced inhibition of ICa, and application of phorbol ester (a protein kinase C activator) (phorbol-12,13-dibutyrate, 0.2 microM) did not mimic the AA action. Some other cis-unsaturated fatty acids (palmitoleic, linoleic, and oleic acids) were also found to depress ICa, while a trans-unsaturated fatty acid (linolelaidic acid) and saturated fatty acids (capric, lauric, myristic, and palmitic acids) had no inhibitory effects on ICa. Myristic acid consistently increased the amplitude of ICa at negative membrane potentials. The present results suggest the possible role of AA, and perhaps other fatty acids, in the physiological and/or pathological modulation of ICa in smooth muscle.

Animals↗

Backfat fatty acid evolution in swine fed diets high in either cis-monounsaturated, trans, or (n-3) fats.

To evaluate the effects of dietary fats on the evolution of the fatty acid profile of swine backfat, 30 castrated Landrace x Duroc pigs averaging 26 kg were assigned three diets with 4% added pomace oil (O), hydrogenated fat (H), or linseed oil (L). Subcutaneous fat samples were taken from biopsies at 0, 17, 31, and 60 d, and at 24 h postmortem when pigs averaged 95 kg live weight at 82 d on trial. On d 17, saturated fatty acid (SFA) content was 2% lower for the O diet than for H and L (P < .089). There was a linear increase in SFA at a monthly rate of 2% in the three diets. The increase was mainly due to 18:0; palmitic acid percentages showed no variation. Pigs fed the O diet experienced exponential increases in monounsaturated fatty acids (MUFA), especially until d 17 (P < .001). In pigs fed the H or L diets, MUFA contents decreased at a monthly rate of 2.40%, and the same happened with oleic acid contents. Diets rich in (n-3) (L) and trans-fatty acids (H) caused exponential increases in swine backfat contents of total (n-3) and total trans, respectively. At d 31 (P < .001), 72 to 73% of the maximum contents had been reached. Contents for 20:3(n-3), 20:5(n-3), and 22:5(n-3) follow a pattern similar to that of their precursor 18:3(n-3), showing an exponential increase in pigs fed the L diet, and contents for H and O treatments were lower (P < .001). The three diets caused a linear decrease in (n-6) fatty acid contents throughout the 82-d trial.

Adipose Tissue↗

Effects of fatty acid substitution on the release of enzymes by osmotic shock.

The release of enzymes by osmotic shock from Escherichia coli strain 30E, an unsaturated fatty acid auxotroph, was examined in culture supplemented with either cis- or trans-unsaturated fatty acids. Cultures grown in oleate-supplemented medium release a large fraction of the total cyclic phosphodiesterase, acid hexose phosphatase, and 5'-nucleotidase following osmotic shock. Cultures grown in elaidate-supplemented medium release much less of these same enzymes after shock treatment. Cultures grown with either supplementation show total release of these enzymes upon conversion to spheroplasts, demonstrating that the enzymes are in the periplasmic space in both cases. Cultures grown with either oleate or elaidate as fatty acid source were washed and suspended in medium containing the other isomer. The change from oleate to elaidate resulted in a rapid decrease in ability of the cells to release the three enzymes after osmotic shock so that within a 25% increase in cell mass the culture responded to osmotic shock as would a culture grown overnight in elaidate-supplemented medium. The reverse experiment resulted in a gradual increase in the ability of the cells to respond to osmotic shock. The outer membrane of E. coli is altered by the incorporation of elaidate, as indicated by electron microscopic data.

Arginine↗

Influence of dietary fat type on arterial thrombosis tendency.

Cardiovascular disease has a multifactorial aetiology, as is illustrated by the existence of numerous risk indicators, many of which can be influenced by the dietary fat type. It should be recalled, however, that only after a cause-and-effect relationship has been established between the disease and a given risk indicator (called a risk factor in that case), modifying this factor can be expected to affect disease morbidity and mortality. In this review, effects of dietary lipids on cardiovascular risk are considered, with special emphasis on modification of arterial thrombosis and platelet thrombotic processes, coagulation and fibrinolysis. Although epidemiological studies do not give entirely consistent results, replacement of dietary saturated by unsaturated fatty acids generally lowers cardiovascular morbidity and mortality. The few (secondary) prevention studies reported so far confirmed this for fish oil or fish oil concentrates, as well as for vegetable oils rich in oleic-, linoleic- or a-linolenic acids. Animal thrombosis models demonstrated that dietary unsaturated fatty acids reduce arterial thrombosis tendency as compared to saturated fatty acids. Using restenosis after percutaneous transluminal coronary angioplasty (PTCA) and occlusion of coronary artery bypass grafts (CABG) as human models of arterial thrombosis, fish oil treatment has been shown to have an anti-thrombotic effect, but results are inconsistent (PTCA) or need confirmation (CABG). Major thrombotic and anti-thrombotic conditions and processes include endothelial integrity, thrombotic functions of blood platelet, coagulation, and fibrinolysis. The dietary fat type can affect endothelial integrity, but results are inconsistent and often difficult to interpret in terms of arterial thrombosis tendency. The same can be concluded for platelet aggregation, especially because results of dietary interventions often depend on the aggregation measuring technique. Novel well-validated methods are required to solve this problem. Dietary fats can affect certain factors involved in blood coagulation and fibrinolysis. Thus, Factor VII activity is increased by the fat content of the diet. However, the dietary fat type seems of less importance. Studies addressing the effect of specific fatty acids on extrinsic clotting and thrombin formation in vivo did not produce consistent results. The same holds for the dietary fatty acid effects on promoters and inhibitors of the plasma fibrinolytic potential. Although trans unsaturated fatty acids may increase cardiovascular risk, this is probably not mediated by effects on arterial thrombosis.

Animals↗