Substrate specificity of <Emphasis Type="Italic">Stenotrophomonas nitritireducens</Emphasis> in the hydroxylation of unsaturated fatty acid

An isolated bacterium that converted unsaturated fatty acids to hydroxy fatty acids was identified as Stenotrophomonas nitritireducens by API analysis, cellular fatty acids compositions, sequencing the full 16S ribosomal ribonucleic acid, and evaluating its nitrite reduction ability. S. nitritireducens has unique regio-specificity for C16 and C18 cis-9 unsaturated fatty acids. These fatty acids are converted to their 10-hydroxy fatty acids without detectable byproducts. Among the cis-9-unsaturated fatty acids, S. nitritireducens showed the highest specificity for linoleic acid. The cells converted 20 mM linoleic acid to 13.5 mM 10-hydroxy-12(Z)-octadecenoic acid at 30°C and pH 7.5 with a yield of 67.5% (mol/mol).     

Metabolic Partitioning of Endogenous Fatty Acid in Adipocytes

Objective: To develop an accurate new method to measure the partitioning of adipocyte endogenous fatty acids among different metabolic pathways, a critical step toward understanding the regulatory mechanism by which fat disposition is modulated. Research Methods and Procedures: Isolated primary rat adipocytes were pre‐incubated with isotope‐labeled fatty acids. This allows determination of the specific activity of labeled fatty acids in the endogenous lipid pool. After the removal of exogenous fatty acids, the disposition of endogenous fatty acids into the three major metabolic pathways, namely, oxidation, re‐esterification, and release into the medium, was measured independently. This was compared with the total lipolytic release of endogenous fatty acids, as measured by glycerol release. Adipocytes from normal fed and fasted animals were used to determine the effects of physiological variations on the metabolic fate of endogenous fatty acids. Results: In normal fed animals, 0.2% of endogenous fatty acids were oxidized, 50.1% were released, and 49.7% were re‐esterified. Fasting doubled the partitioning of fatty acids toward oxidation (p < 0.05) in association with increased lipolysis (1.4‐fold increase) (p < 0.05). This effect was completely abolished by the addition of insulin to the cells (61% reduction) (p < 0.05). Discussion: The endogenous fatty acids in adipocytes are actively oxidized. This process can be regulated by altered physiological conditions or by insulin. Over time, it is possible that a small shift of fatty acids toward oxidation could have a significant impact on body fuel economy. This hypothesis needs to be tested.     

Influence of salt concentration and temperature on the fatty acid compositions of Ectothiorhodospira and other halophilic phototrophic purple bacteria

Influences of the salt concentration on the fatty acid composition of Ectothiorhodospira species and other phototrophic purple bacteria have been analysed. Major fatty acids in bacteria of the genera Rhodobacter, Rhodopseudomonas, Chromatium, and Ectothiorhodospira were straight chain saturated and monounsaturated C-16 and C-18 fatty acids. Salt-dependent responses of all investigated bacteria revealed relations to their salt optima. Minimum values of C-16 and saturated fatty acids and maximum values of C-18 and unsaturated fatty acids were found at or close to the salt optima. Responses of Ectothiorhodospira mobilis upon changes in salinity were nearly identical, whether cells were grown in batch culture or in continuous culture with identical dilution rates at all salt concentrations. With increasing temperature, the fatty acid composition of Ectothiorhodospira mobilis and Ectothiorhodospira halophila strains showed decreasing portions of C-18 and of unsaturated fatty acids, while the contents of C-16 and saturated fatty acids increased. The results are discussed with respect to bilayer stabilisation and membrane fluidity.Abbreviations PC phosphatidylcholine - PG phosphatidylglycerol - CL cardiolipin - PE phosphatidylethanolamine     

Changes in fatty acid branching and unsaturation ofStreptomyces cinnamonensis as a response to NaCl concentration

Three differentStreptomyces cinnamonensis strains (viz. standard, regulatory, and non-differentiating) were grown in a synthetic medium containing 0–4% NaCl and the composition of total cellular fatty acids was analyzed. The increasing salt concentration resulted in an increasing proportion of even-numbered straight-chain fatty acids, mostly at the expense of branched-chain fatty acids. Unsaturated fatty acids, palmitoleic acid in particular, represent the major proportion of straight-chain fatty acids.     

Mass Spectrometry in Carbohydrate Chemistry

In this study, the physiological effects of fatty acids with conjugated double bonds were widely examined in vitro and in vivo. Initially, a method for determination of conjugated fatty acids in food and biological samples was established. I then clarified that the oxidative stability of conjugated fatty acids was improved by the form of triacylglycerol and addition of an antioxidant, and the influence of this effect on the metabolism and pharmacokinetics of conjugated fatty acids was clarified in vivo. In addition, antitumor, anti-angiogenesis, and antiobesity effects of conjugated fatty acids were found for the first time, thus demonstrating the usefulness of conjugated fatty acids. This communication mainly outlines the data obtained for conjugated linolenic acid. In addition, this review summarizes my research on conjugated fatty acid.     

Fatty Acid Specificity in Lipase-Catalyzed Synthesis of Glucoside Esters

The fatty acid specificity of the B-lipase derived from Candida antarctica was investigated in the synthesis of esters of ethyl D-glucopyranoside. The specificity was almost identical with respect to straight-chain fatty acids with 10 to 18 carbon atoms. However, lower fatty acids such as hexanoic and octanoic acid and the unsaturated 9-cis-octadecenoic acid were found to be poor substrates of the enzyme. As a consequence of this selectivity, these fatty acids were accumulated in the unconverted fraction when ethyl D-glucopyranoside was esterified with an excess of a mixture of fatty acids. This accumulation can reduce the overall effectiveness of the process as the activity of the lipase was found to be reduced when exposed to high concentrations of short-chain fatty acids. Finally, using a simplified experimental set-up, the specificity of the C. antarctica B-lipase was compared to the specificity of lipases derived from C. rugosa, Mucor miehei, Humicola, and Pseudomonas. Apart from the C. rugosa lipase, which exhibited a very poor performance, all the enzymes showed a very similar specificity with respect to fatty acids longer than octanoic acid while only the C. antarctica B-lipase showed activity towards sort-chain fatty acids.     

Effect of supplementation with exogenous fatty acid on the biological properties of a fatty acid requiring auxotroph ofSalmonella typhimurium

The effects of changes in fatty acid composition of the cell membrane on different biological functions ofSalmonella typhimurium have been studied with the help of a temperature sensitive fatty acid auxotroph which cannot synthesise unsaturated fatty acids at high temperature. On being shifted to nonpermissive temperature the cells continue growing for another one and half to two generations. The rates of protein and DNA syntheses run parallel to the growth rate but the rate of RNA synthesis is reduced. Further, there is a gradual reduction in the rate of transport of exogenous uridine and thymidine into the soluble pool. The transport process can be restored by supplementing the growth medium with cis-unsaturated fatty acids but not trans-unsaturated ones although the growth of the cells is resumed by supplementation with eithercis or trans-unsaturated fatty acids. However, supplementation withtrans, trans-unsaturated fatty acids leads to only partial recovery of the transport process. The rate of oxygen uptake is also affected in cells grown in the presence of thetrans-unsaturated fatty acids, elaidic acid and palmitelaidic acid. Analysis of cells grown under different fatty acid supplementation indicate that fatty acid composition of the cell membrane, especially the ratio of unsaturated to saturated fatty acids varies with temperature shift and supplementation of the growth media with fatty acids.     

Physiological state of microbial communities and mats of the bottom sediments in the marine shallow-water hydrothermal ecosystem of Kraternaya Bight (Kurile Islands)

The physiological state of littoral and sublittoral microbial communities in a marine shallow-water hydrothermal ecosystem (Kraternaya Bight) was studied using lipid biomarkers. The ratio trans/cis (n-7) isomers of monoenic fatty acids (FAs) of polar lipids in intertidal and subtidal algobacterial and bacterial mats of the bight exceeded 0.1 significantly; this indicated a stress state in bacteria. No concomitant increase was found in the ratio of cyclopropane fatty acids to 16: 1 and 18: 1 (n-7) cis monoenic fatty acids. In bottom sediments, the ratio trans/cis (n-7) isomers of monoenic fatty acids was below 0.1. A positive correlation (r = 0.71) was revealed between the ratio trans/cis isomers of (n-7) monoenic fatty acids and the content of saturated fatty acids.     

Competition between cis,trans and Cyclopropane Fatty Acid Formation and its Impact on Membrane Fluidity

The impact of cis, trans and cyclopropane fatty acids on membrane fluidity was investigated using batch‐grown Pseudomonas putida P8 and Comamonas testosteroni ATCC 17454. A major difference observed between the two investigated strains is the absence of the ability to synthesize trans‐unsaturated fatty acids in Comamonas. When grown exponentially at 30 °C, a shift to 35 °C increased the trans/cis ratios of the fatty acids of P. putida P8 from 0 to 0.81 and 0 to 1.07, in lipid extracts and cell hydrolyzates, respectively. After prolonged growth followed by nutrient deprivation for 48 h, both at 30 °C, trans fatty acids were absent, but the cyclo/cis ratios rose from 0.1 to 1.55 in lipid extracts, and from 0.1 to 1.54 in cell hydrolyzates. C. testosteroni ATCC 17454 contained no cyclo fatty acids when harvested in the exponential phase after 6 h, whereas after 72 h cultivation, the cyclo/cis ratios rose to 0.49 and 0.47, in lipid extracts and cell hydrolyzates, respectively. Trans fatty acids were never observed in this strain. Increased cyclo/cis and trans/cis ratios correlated with decreased fluidity measured by the fluorescence anisotropy of 1,6‐diphenyl‐1,3,5‐hexatriene (DPH) intercalated in the bilayers of liposomes and by Fourier Transform Infrared (FTIR) spectroscopy of lipids prepared from the cells. The specific effect of cyclopropane fatty acids on membrane fluidity was much smaller than that of trans fatty acids. FTIR‐measurements of intact cells of P. putida P8 confirmed the high potency of trans fatty acids to decrease the fluidity. In cells with induced cyclopropane fatty acid synthesis, the membranes remained more fluidized, indicating the lower importance of these fatty acids for homeoviscosis.     

Comparative Study of Fatty Acids Profile in Eleven Wild Mushrooms of Boletacea and Russulaceae Families

Eleven species of wild mushrooms which belong to Boletaceae and Russulaceae families were examined by gas chromatography (GC) and gas chromatography–mass spectrometry (GC/MS) analysis for the presence of fatty acids. As far as we know, the fatty acid profiles of B. purpureus and B. rhodoxanthus were described for the first time. Twenty‐six fatty acids were determined. Linoleic (19.5 – 72%), oleic (0.11 – 64%), palmitic (5.9 – 22%) and stearic acids (0.81 – 57%) were present in the highest contents. In all samples, unsaturated fatty acids dominate. Agglomerative hierarchical clustering was used to display the correlation between the fatty acids and their relationships with the mushroom species. Based on the fatty acids profile in the samples, the mushrooms can be divided into two families: Boletaceae and Russulaceae families, using cluster analysis.     

Fatty acid composition of wild Odontesthes bonariensis (Valenciennes 1835) larvae: implications on lipid metabolism and trophic relationships

The fatty acid composition of the pejerrey Odontesthes bonariensis larvae was studied to elucidate potential dietary relationships. Their principal fatty acids were characteristic for membrane lipids of aquatic organisms. The fatty acid composition varied little throughout seasons, with high proportions of 22 : 6(n‐3) (27% of total fatty acids), which is biosynthesized de novo from dietary precursor fatty acids and/or accumulated from the diet. Other major fatty acids were 16 : 0 and 18 : 0. The diatom‐typical 16 : 1(n‐7) and other dietary fatty acids (zooplankton and microplankton) are not reflected in the larvae, thus limiting the use of fatty acids as trophic markers for food web relationships of atherinopsids.     

Analysis of whole cellular fatty acids and anastomosis relationships of binucleate Rhizoctonia spp. associated with Ceratobasidium cornigerum

Previous research has demonstrated that whole cellular fatty acids analysis is a useful tool for identifying and establishing taxonomic relationships between anastomosis groups (AGs) and related Rhizoctonia isolates. In this experiment, the composition of fatty acid of 28 isolates of teleomorph genus Ceratobasidium cornigerum, consisting of binucleate Rhizoctonia, AG-A, AG-B(o), AG-C, AG-P, and AG-Q, was evaluated using gas chromatography. Eleven fatty acids identified, i.e., myristic, pentadecanoic, palmitic, 2-hydroxypalmitic, palmitoleic, heptadecanoic, 9-heptadecenoic, stearic, oleic, linoleic, and linolenic acids, were present in isolates of AG-A, AG-B(o), AG-C, AG-P, and AG-Q. The major fatty acids, palmitic, oleic, and linoleic acids, were common in all isolates, constituting 87.1% to 94.7% of the whole cellular fatty acids identified. Isolates within the same AG were closely clustered, whereas isolates from different AGs were clearly and distinctly clustered based on average linkage cluster analysis of whole cellular fatty acids. Principal-component analysis generated from all fatty acids also confirmed the divergent separation of the 5 AGs of binucleate Rhizoctonia.     

Unsaturated free fatty acids regulate cAMP relay and cell-cell adhesion duringDictyostelium discoideum aggregation

Summary Cell-cell adhesion and cAMP-stimulated cAMP production (cAMP relay) regulate the aggregation that occurs early in theDictyostelium discoideum developmental cycle. Increasing the concentration of neutral lipids inD. discoideum membranes inhibits both cell-cell adhesion and cAMP relay. Fractionation experiments revealed that it was the free fatty acids, one group of the neutral lipids, that inhibited both cAMP relay and cell-cell adhesion. Work with commercially-available free fatty acids demonstrated that the addition of saturated free fatty acids, palmitic acid and stearic acid, did not alter cell-cell adhesion or cAMP relay. The addition of unsaturated free fatty acids inhibited both cAMP relay and cell-cell adhesion in a dose-dependent, saturable manner. To test the physiological significance of these observations, the concentrations of endogenous unsaturated fatty acids were modified by altering theD. discoideum diet. Decreasing the amount of fatty acids 18 carbons long with 2 double bonds enhanced cAMP relay and cell-cell adhesion. These results suggest that fatty acids may be important regulators of cAMP responsiveness and cell-cell adhesion duringD. discoideum aggregation, and therefore may play important roles in development.     

The synthesis of astaxanthin esters,independent of the formation of cysts,highly correlates with the synthesis of fatty acids in <Emphasis Type="Italic">Haematococcus pluvialis</Emphasis>

The compositions and contents of astaxanthin esters and fatty acids in four types of Haematococcus pluvialis cells were studied by HPLC and GC-MS. Results showed that the synthesis and accumulation of astaxanthin was independent of the formation of cysts, but was highly correlated with the synthesis and accumulation of fatty acids, though it is an well known phenomenon that the accumulation of astaxanthin is usually accompanied by the formation of cyst. The red cysts contain more than 30% of fatty acids, with 81% of the unsaturated fatty acids. Taken together, besides a resource of astaxanthin, H. pluvialis would be a good resource of valuable fatty acids.     

Effect ofcis andtrans unsaturated fatty acids on the transport properties ofSalmonella typhimurium

The transport of α-methyl-D-glucoside and two aminoacids, L-phenylalanine and L-leucine by a temperature sensitive fatty acid requiring mutant ofSalmonella typhimurium was studied under conditions of supplementation withcis or trans-unsaturated fatty acids. The results of such experiments definitely establish a relationship between the fatty acids composition of the membrane and the transport property of the cells. Cells grown in the presence of trans-unsaturated fatty acids cannot transport so efficiently as compared to the cis-unsaturated fatty acid-grown cells except linolelaidic acid, atrans-trans-unsaturated fatty acid. Protein: phospholipid ratio of the membrane also varies significantly under such conditions. The affinity of L-phenylalanine transport carrier for the substrate changes remarkably in cells grown in the presence of differentcis or trans-unsaturated fatty acids and indicate the possible role of membrane lipids in membrane assembly as well as regulation of the activity of L-phenylalanine transport system.     

The impact of dietary fats,photoperiod, temperature and season on morphological variables,torpor patterns,and brown adipose tissue fatty acid composition of hamsters,Phodopus sungorus

We investigated how dietary fats and oils of different fatty acid composition influence the seasonal change of body mass, fur colour, testes size and torpor in Djungarian hamsters, Phodopus sungorus, maintained from autumn to winter under different photoperiods and temperature regimes. Dietary fatty acids influenced the occurrence of spontaneous torpor (food and water ad libitum) in P. sungorus maintained at 18°C under natural and artificial short photoperiods. Torpor was most pronounced in individuals on a diet containing 10% safflower oil (rich in polyunsaturated fatty acids), intermediate in individuals on a diet containing 10% olive oil (rich in monounsaturated fatty acids) and least pronounced in individuals on a diet containing 10% coconut fat (rich in saturated fatty acids). Torpor in P. sungorus on chow containing no added fat or oil was intermediate between those on coconut fat and olive oil. Dietary fatty acids had little effect on torpor in animals maintained at 23°C. Body mass, fur colour and testes size were also little affected by dietary fatty acids. The fatty acid composition of brown fat from hamsters maintained at 18°C and under natural photoperiod strongly reflected that of the dietary fatty acids. Our study suggests that the seasonal change of body mass, fur colour and testes size are not significantly affected by dietary fatty acids. However, dietary fats influence the occurrence of torpor in individuals maintained at low temperatures and that have been photoperiodically primed for the display of torpor.Abbreviations BAT brown adipose tissue - bm body mass - FA fatty acid(s) - MR metabolic rate - MUFA monounsaturated fatty acid(s) - PUFA polyunsaturated fatty acid(s) - SFA saturated fatty acid(s) - T _a air temperature - T _b body temperature - T_s body surface temperature(s) - TNZ thermoneutral zone - UFA unsaturated fatty acid(s)     

Effect of frozen storage on fatty acid composition of the different tissues of four scombrid and one dussumeriid species

In the current study, the effect of frozen storage at ?18°C was evaluated on fatty acid composition of different body parts (liver, muscle tissue, and viscera) of narrow‐barred Spanish mackerel (Scomberomorus commerson, Lacépède, 1800), longtail tuna (Thunnus tonggol, Bleeker, 1851), kawakawa (Euthynnus affinis, Cantor, 1849), king mackerel (Scomberomorus guttatus, Bloch & Schneider, 1801), and rainbow sardine (Dussumieria acuta, Valenciennes, 1847) caught in the Persian Gulf. Changes in saturated fatty acids (SFAs), monounsaturated fatty acids (MUFAs), polyunsaturated fatty acids (PUFAs), eicosapentaenoic acid plus docosahexaenoic acid/palmitic acid (EPA+DHA/C16), ω3 PUFA/ω6 PUFA (ω3/ω6), and polyunsaturated fatty acids/saturated fatty acids (PUFA/SFA) were investigated during a 6‐month period. A decrease in unsaturated fatty acids, particularly PUFAs (60–100%) as well as ω3/ω6, EPA+DHA/C16 (polyene index) and PUFA/SFA ratios, indicated a decrease in the nutritional values of the samples.     

Die Gefriertrocknung von Rinderkot

Fatty acids in fish can arise from two sources: synthesis de novo from non‐lipid carbon sources within the animal, or directly from dietary lipid. Acetyl‐CoA derived mainly from protein can be converted to saturated fatty acids via the combined action of acetyl‐CoA carboxylase and fatty acid synthetase. The actual rate of fatty acid synthesis de novo is inversely related to the level of lipid in the diet. Freshwater fish can de‐saturate endogenously‐synthesized fatty acids to monounsaturated fatty acids via a A9 desaturase but lack the necessary enzymes for complete de novo synthesis of polyunsaturated fatty acids which must therefore be obtained preformed from the diet. Most freshwater fish species can desaturate and elongate 18:2(n‐6) and 18:3(n‐3) to their C20 and C22 homologues but the pathways involved remain ill‐defined. Cyclooxygenase and lipoxygenase enzymes can convert C20 polyunsaturated fatty acids to a variety of eicosanoid products. The dietary ratio of (n‐3) to (n‐6) polyunsaturated fatty acids influences the pattern of eicosanoids formed. The ß‐oxidation of fatty acids can occur in both mitochondria and peroxisomes but mi‐tochondrial ß‐oxidation is quantitatively more important and can utilise a wide range of fatty acid substrates.     

Reliable identification of Prevotella and Butyrivibrio spp. from rumen by fatty acid methyl ester profiles

Data for bacterial identification were provided by culturing anaerobic bacteria under standardized conditions followed by extraction and methylation of cellular long-chain fatty acids and gas chromatographic analysis. The databases of fatty acid methyl ester (FAMEs) profiles for two predominant ruminal genera,Prevotella andButyrivibrio, were created. Major long-chain cellular fatty acids found in the 23 analyzedPrevotella strains were 15:0 (anteiso), 15:0, 15:0 (iso) and 16:0. The strains ofPrevotella could be well identified on species level by the characteristic ratios among major fatty acids and by acids unique fatty for each species. The 45Butyrivibrio strains were grouped into 4 major and 2 minor groups according to FAMEs profiles. The major fatty acids for the bulk of theButyrivibrio strains were 14:0, 15:1, 16:0 and 16:0 (iso). This groups corresponded to those based on 16S rDNA sequences.     

The degree of dietary fatty acid unsaturation affects torpor patterns and lipid composition of a hibernator

Diets rich in unsaturated and polyunsaturated fatty acids have a positive effect on mammalian torpor, whereas diets rich in saturated fatty acids have a negative effect. To determine whether the number of double bonds in dietary fatty acids are responsible for these alterations in torpor patterns, we investigated the effect of adding to the normal diet 5% pure fatty acids of identical chain length (C18) but a different number of double bonds (0, 1, or 2) on the pattern of hibernation of the yellow-pine chipmunk, Eutamias amoenus. The response of torpor bouts to a lowering of air temperature and the mean duration of torpor bouts at an air temperature of 0.5°C (stearic acid C18:0, 4.5±0.8 days, oleic acid C18:1, 8.6±0.5 days; linoleic acid C18:2, 8.5±0.7 days) differed among animals that were maintained on the three experimental diets. The mean minimum body temperatures (C18:0, +2.3±0.3°C; C18:1, +0.3±0.2°C; C18:2,-0.2±0.2°C), which torpid individuals defended by an increase in metabolic rate, and the metabolic rate of torpid animals also differed among diet groups. Moreover, diet-induced differences were observed in the composition of total lipid fatty acids from depot fat and the phospholipid fatty acids of cardiac mitochondria. For depot fat 7 of 13 and for heart mitochondria 7 of 14 of the identified fatty acids differed significantly among the three diet groups. Significant differences among diet groups were also observed for the sum of saturated, unsaturated and polyunsaturated fatty acids. These diet-induced alterations of body fatty acids were correlated with some of the diet-induced differences in variables of torpor. The results suggest that the degree of unsaturation of dietary fatty acids influences the composition of tissues and membranes which in turn may influence torpor patterns and thus survival of hibernation.Abbreviations bm body mass - T _a air temperature - T _b body temperature - FA fatty acid - MR metabolic rate - MUFA monounsaturated fatty acids - PUFA polyunsaturated fatty acids - VO₂ rate of oxygen consumption - SFA saturated fatty acids - UFA unsaturated fatty acids - UI unsaturation index - SNK Student-Newman-Keuls test