Changes in the composition of intracellular lipids and hydrocarbons in Candida tropicalis in the absence of growth]

The fatty acid composition of lipids and the composition of hydrocarbons were studied in Candida tropicalis cultivated on a medium with propionic acid and incubated in the conditions of starvation and on a medium containing glucose-1-6-14C but no nitrogen. Intracellular fatty acids with an odd number of carbon atoms were found to be easily metabolized by yeast cells. The content and composition of intracellular hydrocarbons were very labile and underwent intensive transformation caused by changes in the metabolism of the yeast cell.     

Nutritional Alteration of the Fatty Acid Composition of a Thermophilic Bacillus Species

The fatty acid composition of a thermophilic Bacillus sp. was altered by the addition of isobutyrate, isovalerate, alpha-methylbutyrate, leucine, and isoleucine to the growth medium. With isobutyrate, 81% of the fatty acids had 16 carbon atoms and 79% were iso-fatty acids with an even number of carbon atoms. With leucine, 58% of the fatty acids had 15 carbon atoms and 86% were iso-fatty acids with an odd number of carbon atoms. With isoleucine, 72% of the fatty acids had 17 carbon atoms and 88% were anteiso-fatty acids with an odd number of carbon atoms. Thus, by altering the composition of the growth medium, cells were produced in which the majority of the fatty acids had either 15, 16, or 17 carbons and belonged to each of the three groups of branched-chain fatty acids. The wide variation observed in the fatty acid composition makes it unlikely that any specific branched-chain fatty acid is required for vital functions.     

Anaerobic n-alkane metabolism by a sulfate-reducing bacterium, Desulfatibacillum aliphaticivorans strain CV2803T

The alkane-degrading, sulfate-reducing bacterium Desulfatibacillum aliphaticivorans strain CV2803T, recently isolated from marine sediments, was investigated for n-alkane metabolism. The total cellular fatty acids of this strain had predominantly odd numbers of carbon atoms (C odd) when the strain was grown on a C-odd alkane (pentadecane) and even numbers of carbon atoms (C even) when it was grown on a C-even alkane (hexadecane). Detailed analyses of those fatty acids by gas chromatography/mass spectrometry allowed us to identify saturated 2-, 4-, 6-, and 8-methyl- and monounsaturated 6-methyl-branched fatty acids, with chain lengths that specifically correlated with those of the alkane. Growth of D. aliphaticivorans on perdeuterated hexadecane demonstrated that those methyl-branched fatty acids were directly derived from the substrate. In addition, cultures on pentadecane and hexadecane produced (1-methyltetradecyl)succinate and (1-methylpentadecyl)succinate, respectively. These results indicate that D. aliphaticivorans strain CV2803T oxidizes n-alkanes into fatty acids anaerobically, via the addition of fumarate at C-2. Based on our observations and on literature data, a pathway for anaerobic n-alkane metabolism by D. aliphaticivorans is proposed. This involves the transformation of the initial alkylsuccinate into a 4-methyl-branched fatty acid which, in addition to catabolic reactions, can alternatively undergo chain elongation and desaturation to form storage fatty acids.     

Stability of the fatty acid composition of actinomycetes

The stability of fatty acid composition of total extractable lipids was studied in Streptomyces cultures. The type of fatty acid composition typical of the Streptomyces genus remains stable when the actinomycetes were grown as submerged cultures in various synthetic media: saturated fatty acids with methyl branching in the chain predominated in all of the cases, and fatty acids with an uneven number of carbon atoms in the chain prevailed in most of the cases. Fatty acids with the anteiso structure predominated among the acids with a branched chain, amounting to more than a half of the latter and reaching sometimes 50% of the total fatty acid content. Methyl branchings were located in the anteiso position in fatty acids with an uneven number of carbon atoms, and in the iso position in fatty acids with an even number of carbons. Unsaturated fatty acids were found as a minor component.     

Anaerobic transformation of alkanes to fatty acids by a sulfate-reducing bacterium,strain Hxd3

Strain Hxd3, an alkane-degrading sulfate reducer previously isolated and described by Aeckersberg et al. (F. Aeckersberg, F. Bak, and F. Widdel, Arch. Microbiol. 156:5-14, 1991), was studied for its alkane degradation mechanism by using deuterium and (13)C-labeled compounds. Deuterated fatty acids with even numbers of C atoms (C-even) and (13)C-labeled fatty acids with odd numbers of C atoms (C-odd) were recovered from cultures of Hxd3 grown on perdeuterated pentadecane and [1,2-(13)C(2)]hexadecane, respectively, underscoring evidence that C-odd alkanes are transformed to C-even fatty acids and vice versa. When Hxd3 was grown on unlabeled hexadecane in the presence of [(13)C]bicarbonate, the resulting 15:0 fatty acid, which was one carbon shorter than the alkane, incorporated a (13)C label to form its carboxyl group. The same results were observed when tetradecane, pentadecane, and perdeuterated pentadecane were used as the substrates. These observations indicate that the initial attack of alkanes includes both carboxylation with inorganic bicarbonate and the removal of two carbon atoms from the alkane chain terminus, resulting in a fatty acid one carbon shorter than the original alkane. The removal of two terminal carbon atoms is further evidenced by the observation that the [1,2-(13)C(2)]hexadecane-derived fatty acids contained either two (13)C labels located exclusively at their acyl chain termini or none at all. Furthermore, when perdeuterated pentadecane was used as the substrate, the 14:0 and 16:0 fatty acids formed both carried the same numbers of deuterium labels, while the latter was not deuterated at its carboxyl end. These observations provide further evidence that the 14:0 fatty acid was initially formed from perdeuterated pentadecane, while the 16:0 fatty acid was produced after chain elongation of the former fatty acid with nondeuterated carbon atoms. We propose that strain Hxd3 anaerobically transforms an alkane to a fatty acid through a mechanism which includes subterminal carboxylation at the C-3 position of the alkane and elimination of the two adjacent terminal carbon atoms.     

Anaerobic n-Alkane Metabolism by a Sulfate-Reducing Bacterium, Desulfatibacillum aliphaticivorans Strain CV2803T

The alkane-degrading, sulfate-reducing bacterium Desulfatibacillum aliphaticivorans strain CV2803^T, recently isolated from marine sediments, was investigated for n-alkane metabolism. The total cellular fatty acids of this strain had predominantly odd numbers of carbon atoms (C odd) when the strain was grown on a C-odd alkane (pentadecane) and even numbers of carbon atoms (C even) when it was grown on a C-even alkane (hexadecane). Detailed analyses of those fatty acids by gas chromatography/mass spectrometry allowed us to identify saturated 2-, 4-, 6-, and 8-methyl- and monounsaturated 6-methyl-branched fatty acids, with chain lengths that specifically correlated with those of the alkane. Growth of D. aliphaticivorans on perdeuterated hexadecane demonstrated that those methyl-branched fatty acids were directly derived from the substrate. In addition, cultures on pentadecane and hexadecane produced (1-methyltetradecyl)succinate and (1-methylpentadecyl)succinate, respectively. These results indicate that D. aliphaticivorans strain CV2803^T oxidizes n-alkanes into fatty acids anaerobically, via the addition of fumarate at C-2. Based on our observations and on literature data, a pathway for anaerobic n-alkane metabolism by D. aliphaticivorans is proposed. This involves the transformation of the initial alkylsuccinate into a 4-methyl-branched fatty acid which, in addition to catabolic reactions, can alternatively undergo chain elongation and desaturation to form storage fatty acids.     

Anaerobic Transformation of Alkanes to Fatty Acids by a Sulfate-Reducing Bacterium, Strain Hxd3

Strain Hxd3, an alkane-degrading sulfate reducer previously isolated and described by Aeckersberg et al. (F. Aeckersberg, F. Bak, and F. Widdel, Arch. Microbiol. 156:5-14, 1991), was studied for its alkane degradation mechanism by using deuterium and ¹³C-labeled compounds. Deuterated fatty acids with even numbers of C atoms (C-even) and ¹³C-labeled fatty acids with odd numbers of C atoms (C-odd) were recovered from cultures of Hxd3 grown on perdeuterated pentadecane and [1,2-¹³C₂]hexadecane, respectively, underscoring evidence that C-odd alkanes are transformed to C-even fatty acids and vice versa. When Hxd3 was grown on unlabeled hexadecane in the presence of [¹³C]bicarbonate, the resulting 15:0 fatty acid, which was one carbon shorter than the alkane, incorporated a ¹³C label to form its carboxyl group. The same results were observed when tetradecane, pentadecane, and perdeuterated pentadecane were used as the substrates. These observations indicate that the initial attack of alkanes includes both carboxylation with inorganic bicarbonate and the removal of two carbon atoms from the alkane chain terminus, resulting in a fatty acid one carbon shorter than the original alkane. The removal of two terminal carbon atoms is further evidenced by the observation that the [1,2-¹³C₂]hexadecane-derived fatty acids contained either two ¹³C labels located exclusively at their acyl chain termini or none at all. Furthermore, when perdeuterated pentadecane was used as the substrate, the 14:0 and 16:0 fatty acids formed both carried the same numbers of deuterium labels, while the latter was not deuterated at its carboxyl end. These observations provide further evidence that the 14:0 fatty acid was initially formed from perdeuterated pentadecane, while the 16:0 fatty acid was produced after chain elongation of the former fatty acid with nondeuterated carbon atoms. We propose that strain Hxd3 anaerobically transforms an alkane to a fatty acid through a mechanism which includes subterminal carboxylation at the C-3 position of the alkane and elimination of the two adjacent terminal carbon atoms.     

Lipid fatty acid composition of fungi in the genus Aspergillus grown on media with various sources of nitrogen

The fatty acid composition of lipids synthesized by fungi belonging to the Aspergillus genus was studied during their growth on media containing different organic and inorganic nitrogen sources. On the average, the cultures were shown to accumulate from 7 to 30 g/L of biomass and to synthesize from 3 to 13% of lipids. The lipids were found to contain saturated and unsaturated fatty acids with the number of carbon atoms from 14 to 18. The fungi had a typical fatty acid composition of lipids which did not depend on the composition of the growth medium.     

Identification of the cuticular lipid composition of the Western Flower Thrips Frankliniella occidentalis

The Western Flower Thrips Frankliniella occidentalis effectively resists many insecticides, but it can be controlled by the use of bioinsecticides such as entomopathogenic fungi. The epicuticular chemistry of these insects is therefore of great interest, and accordingly, the cuticular lipid composition of F. occidentalis was analysed. It was found that the cuticular lipids of both the adult and larval stages of F. occidentalis consist of two groups of compounds--hydrocarbons and free fatty acids. The same hydrocarbon pattern was found in both adults and larvae, with the exception of n-hentriacontane, which was detected only in adult insects. The following homologous series were identified: n-alkanes from C-25 to C-29 (31) with the marked dominance of odd numbers of carbon atoms, 3-methylalkanes with 26 and 28 carbon atoms, and branched monomethylalkanes (branched at C-9, -11, -13 and -15) with 26, 28 and 30 carbon atoms. The chemical composition of the free fatty acids consists of two homologous series: saturated (C(14:0), C(16:0), C(18:0)) and unsaturated fatty acids (C(16:1) and C(18:1)). This analysis confirmed the lack of potential inhibitors of entomopathogenic fungi in the cuticular lipids of this insect species.     

Microbial assimilation of hydrocarbons: cellular distribution of fatty acids

The distribution of cellular fatty acids in defined lipid classes was analyzed in Micrococcus cerificans after growth on specified hydrocarbons. Neutral lipid, phospholipid, and cell residue fatty acids were qualitatively and quantitatively determined for M. cerificans grown on nutrient broth, tetradecane (C(14)), pentadecane (C(15)), hexadecane (C(16)), and heptadecane (C(17)), respectively. Percentage of total cellular fatty acid localized in defined lipid classes from cells grown on the above growth substrates was (i) neutral lipid-11.8, 1.81, 7.74, 23.1, and 2%; (ii) phospholipid-74.5, 65, 66.43, 62.1, and 86%; (iii) cell residue lipid-13.5, 33.29, 25.82, 14.78, and 11.9%. Phospholipid fatty acid chain length directly reflected the carbon number of the alkane substrate, with 40, 84, 98, and 77% of the fatty acids being 14, 15, 16, and 17 carbons when cells were grown on C(14), C(15), C(16), and C(17)n-alkanes, respectively. The bound lipids of the cell residue after chloroform-methanol extraction were characterized by 2-hydroxydodecanoic and 2-hydroxytetradecanoic acids plus a broad spectrum of fatty acids ranging from C(10) to C(17) chain length. An increase in total unsaturated fatty acid localized in the phospholipids was noted from cells grown on alkanes greater than 15 carbons long. An extracellular accumulation of free fatty acid (FFA) was demonstrated in hexadecane-grown cultures that was not apparent in non-hydrocarbon-grown cultures. Identification of extracellular FFA demonstrated direct derivation from hexadecane oxidation. Studies supporting inhibition of de novo fatty acid biosynthesis in relationship to extracellular FFA and hexadecane oxidation are described. The ability to alter the fatty acid composition of membrane polar lipids in a predictable manner by the alkane carbon source provides an excellent model system for the investigation of membrane structure-function relationships in M. cerificans.     

Lipid synthesis by Micrococcus freudenreichii in media containing unsaturated hydrocarbon's]

The growth and synthesis of lipids by thermotolerant bacteria Micrococcus freudenreichii K-219 were investigated in the mineral medium containing a mixture of unsaturated (I-) and saturated hydrocarbons. The bacteria utilized primarily I-alkenes. In lipids the predominant fractions were phospholipids (57%) and free fatty acids (20%). The content of waxes which were in significant quantities in n-alkane containing media (9%) was not higher than 0.3% dry matter upon utilization of I-alkenes. There was a certain correlation between carbon atoms of synthesized fatty acids and unsaturated hydrocarbons used. Bacteria utilizing I-alkenes showed no elevated unsaturation of cell lipids as compared to those assimilating n-alkanes. These data give evidence for different pathways of oxidation of alkenes and alkanes by the above microbial strain.     

Initial reactions in anaerobic alkane degradation by a sulfate reducer, strain AK-01

An alkane-degrading, sulfate-reducing bacterial strain, AK-01, isolated from a petroleum-contaminated sediment was studied to elucidate its mechanism of alkane metabolism. Total cellular fatty acids of AK-01 were predominantly C even when it was grown on C-even alkanes and were predominantly C odd when grown on C-odd alkanes, suggesting that the bacterium anaerobically oxidizes alkanes to fatty acids. Among these fatty acids, some 2-, 4-, and 6-methylated fatty acids were specifically found only when AK-01 was grown on alkanes, and their chain lengths always correlated with those of the alkanes. When [1,2-(13)C(2)]hexadecane or perdeuterated pentadecane was used as the growth substrate, (13)C-labeled 2-Me-16:0, 4-Me-18:0, and 6-Me-20:0 fatty acids or deuterated 2-Me-15:0, 4-Me-17:0, and 6-Me-19:0 fatty acids were recovered, respectively, confirming that these monomethylated fatty acids were alkane derived. Examination of the (13)C-labeled 2-, 4-, and 6-methylated fatty acids by mass spectrometry showed that each of them contained two (13)C atoms, located at the methyl group and the adjacent carbon, thus indicating that the methyl group was the original terminal carbon of the [1, 2-(13)C(2)]hexadecane. For perdeuterated pentadecane, the presence of three deuterium atoms, on the methyl group and its adjacent carbon, in each of the deuterated 2-, 4-, and 6-methylated fatty acids further supported the hypothesis that the methyl group was the terminal carbon of the alkane. Thus, exogenous carbon appears to be initially added to an alkane subterminally at the C-2 position such that the original terminal carbon of the alkane becomes a methyl group on the subsequently formed fatty acid. The carbon addition reaction, however, does not appear to be a direct carboxylation of inorganic bicarbonate. A pathway for anaerobic metabolism of alkanes by strain AK-01 is proposed.     

Lipid and fatty acid composition of Eisenia foetida

It is found out that the content of lipids in the biomass of the studied populations of Eisenia foetida is rather high: 2.5-5.2% of the wet mass. The content of phospholipids is 40-55%, C27-sterols--1.5-3.4% of the mass of coarse extracts of lipids. Lipids of tissues contain also 47-54% of saturated (C10-C24) fatty acids as well as to 23% of monoene and to 13% of polyene unsaturated (C14-C22) fatty acids. The acids with the odd number of carbon atoms compose about 25% and acids with a branched carbon chain about 23% of the above percentage. Considerable content of lipids and biologically active fatty acids in tissues of the studied object permits considering it as a promising source of raw materials for production of valuable pharmacological preparations.     

Effect of growth temperature on lipid and fatty acid compositions in the blue-green algae, Anabaena variabilis and Anacystis nidulans.

The lipid composition was affected by growth temperature in Anacystis nidulans, but was not in Anabaena variabilis. A. variabilis contained fatty acids of 18 and 16 carbon atoms, which were localized at 1- and 2-positions, respectively, of the glycerol moiety of lipids. Desaturation of C18 acids was affected by the growth temperature. A. nidulans contained fatty acids of 14, 16 and 18 carbon atoms. Monounsaturated and saturated acids were esterified mainly to 1- and 2-position, respectively. Desaturation and chain length of fatty acids were influenced by the growth temperature. The variations in lipid and fatty acid compositions with the growth temperature are discussed in relation to the growth temperature-dependent shift of thermotropic phase transition temperature of the membrane lipids in the blue-green algae.     

Short chain fatty acid inhibition of rat brain Na-K adenosine triphosphatase

The possibility was considered that the sleep-like state seen after injection of short chain fatty acids salts into animals is a result of inhibition of the sodium-potassium activated ATPase. Tris salts of short chain fatty acids inhibited brain Na-K ATPase activity in vitro at concentrations similar to intravenous levels causing narcosis in vivo. The inhibition depended on the logarithm of the concentration of a given acid. The concentration of acid anion which caused 50 per cent inhibition of the enzyme system (I50) was determined for straight and branched chain acids with 4-12 carbon atoms per molecule. The log of I50 concentrations plotted against the number of carbon atoms in the molecule gave a straight line; the inhibitory capacity of an acid increased by a factor of 2.3 for each--CH2--added to the carbon chain. It is suggested that both fatty acid narcosis and the enzyme inhibition result from fatty acid molecules forming an ordered array along the membrane in association with membrane lipids.     

Production of Lipids and Carotinoids by Yellow Streptomyces (in Russian)

The ability of yellow streptomyces to synthesize lipids and carotinoids was examined. It has been established that the content of lipids in the mycelium varied from 9.7 to 57.1% depending on the species and the strains. Cultures belonging to the species Str. nephtigulosus, Str. ambofaciens, Str. citreus showed carotene-synthesizing abilities. In the content of the lipids and carotinoids of the examined cultures were found our fractions possessing biological activity. The fatty acid content of the examined cultures was characterized by the saturated and unsaturated acids containing an even and odd amount of the carbon atoms (from C₁₁ to C₁₈) and a normal and branched chain (iso-C₁₆ and iso-C₁₇).     

Microbial Incorporation of Fatty Acids Derived From n-Alkanes Into Glycerides and Waxes

When n-alkanes with 13 to 20 carbon atoms were fed to a Nocardia closely related to N. salmonicolor, the produced cellular triglycerides and aliphatic waxes invariably contained fatty acids with an even or an odd number of carbon atoms subject to this feature of the n-alkane substrate. Beta-oxidation and C₂ addition are both operative, as evidenced by the spectra of fatty acids incorporated into the cellular lipid components. There is no distinction in the rate of microbial incorporation of the odd-or even-numbered carbon chains. The fatty acids are apparently directly derived from the long chain n-alkanes, rather than synthesized via the classic C₂-condensation route. The alcohol component of waxes produced by the Nocardia is invariably of the same chain length as the n-alkane substrate.     

Initial Reactions in Anaerobic Alkane Degradation by a Sulfate Reducer, Strain AK-01

An alkane-degrading, sulfate-reducing bacterial strain, AK-01, isolated from a petroleum-contaminated sediment was studied to elucidate its mechanism of alkane metabolism. Total cellular fatty acids of AK-01 were predominantly C even when it was grown on C-even alkanes and were predominantly C odd when grown on C-odd alkanes, suggesting that the bacterium anaerobically oxidizes alkanes to fatty acids. Among these fatty acids, some 2-, 4-, and 6-methylated fatty acids were specifically found only when AK-01 was grown on alkanes, and their chain lengths always correlated with those of the alkanes. When [1,2-¹³C₂]hexadecane or perdeuterated pentadecane was used as the growth substrate, ¹³C-labeled 2-Me-16:0, 4-Me-18:0, and 6-Me-20:0 fatty acids or deuterated 2-Me-15:0, 4-Me-17:0, and 6-Me-19:0 fatty acids were recovered, respectively, confirming that these monomethylated fatty acids were alkane derived. Examination of the ¹³C-labeled 2-, 4-, and 6-methylated fatty acids by mass spectrometry showed that each of them contained two ¹³C atoms, located at the methyl group and the adjacent carbon, thus indicating that the methyl group was the original terminal carbon of the [1,2-¹³C₂]hexadecane. For perdeuterated pentadecane, the presence of three deuterium atoms, on the methyl group and its adjacent carbon, in each of the deuterated 2-, 4-, and 6-methylated fatty acids further supported the hypothesis that the methyl group was the terminal carbon of the alkane. Thus, exogenous carbon appears to be initially added to an alkane subterminally at the C-2 position such that the original terminal carbon of the alkane becomes a methyl group on the subsequently formed fatty acid. The carbon addition reaction, however, does not appear to be a direct carboxylation of inorganic bicarbonate. A pathway for anaerobic metabolism of alkanes by strain AK-01 is proposed.     

Effect of the exogenous lipids of complex media on the fatty acid composition of mycelial microorganisms

The effect of exogenous lipid sources on the composition of fatty acids was studied in actinomycetes of the Streptomyces genus and in fungi belonging to the genera Blakeslea, Cunninghamella and Penicillium. The following sources of exogenous lipids were used: soybean and maize flour, sunflower by-products, chicken droppings, maize extract, yeast extract, peptone, sperm whale fat, sunflower and palm oil. The composition of fatty acids in total extracted lipids of the studied mycelial microorganisms was shown to reflect two processes: lipid synthesis de novo and assimilation of exogenous fatty acids. This fact ought to be taken into account both in the chemotaxonomic interpretation of fatty acid composition and in practical recommendations for the utilization of microbial lipids. It is of particular interest to study the physiological role of exogenous lipid metabolism in the cells of microorganisms.     

The effects of short-chain fatty acids on the neuronal membrane functions ofHelix pomatia. III.22Na efflux from the cells

The effect of short-chain fatty acids on both ouabain-sensitive and ouabain-insensitive fractions of 22Na efflux from the neurons of Helix pomatia was studied. Fatty acids, having fewer than 10 carbon atoms in the hydrocarbon chain, increased the ouabain-sensitive 22Na efflux from the neurons, while fatty acids, having more than 9 carbon atoms, inhibited the 22Na efflux in comparison with that in normal physiological solution. All the fatty acids used had an inhibiting effect on the ouabain-insensitive 22Na efflux from the cells independent on the number of carbon atoms in the hydrocarbon chain. These studies indicate that these short-chain fatty acids can be effective modulators of both ouabain-sensitive and ouabain-insensitive fractions of Na efflux from the cells.