Desaturation System Research Articles

Impact of curcumin on fatty acid metabolism.

Free fatty acids (FFAs) and fatty acid synthesis (FAS) activity have significantly contributed to disease states such as insulin resistance, obesity, type 2 diabetes, myocardial infarction, blood pressure, and several types of cancer. Currently, several treatment options are available for patients with these conditions. Due to safety concerns, adverse effects, limited efficacy, and low tolerability associated with many medications, the identification of novel agents with less toxicity and a more favorable outcome is warranted. Curcumin is a phenolic compound derived from the turmeric plant with various biological activities, including anticarcinogenic, antioxidant, antiinflammatory, and hypolipidemic properties. PubMed, Scopus, and Web of Science were searched up to February 2020 for studies that demonstrated the efficacy and mechanisms of curcumin action on FFAs, FAS, and β-oxidation activity, as well as the desaturation system. Most of the evidence is in-vivo and in-vitro studies that demonstrate that curcumin possesses regulatory properties on FFAs levels through its effects on FAS and β-oxidation activity as well as desaturation system, which could improve insulin resistance, obesity, and other FFAs-related disorders. The present study provides a review of the existing in-vitro, in-vivo, and clinical evidence on the effect of curcumin on FFAs and FAS activity, β-oxidation, and desaturation system.

Lipid characterization of an arachidonic acid-rich oil producing fungus Mortierella alpina

Mortierella alpina has been considered as the most effective producer of arachidonic acid (ARA)-rich oil. It was found that several methods could improve the percentage of ARA in total lipids successfully, as they activated the desaturation system on the endoplasmic reticulum. Additionally, in M. alpina the ARA exists in several forms, such as triacylglycerol (TAG), and diacylglycerol (DAG). These forms are caused by different acyltransferases and they determine the nutrient value of the microbial oil. However, few works revealed detailed fatty acid distribution among lipid classes, which to some extent impeded the accurate regulation in ARA accumulation. Herein, this paper gives information on the accumulation process of main lipid classes and the changes of fatty acid composition in these lipids during ARA accumulation period in M. alpina. The result demonstrates that TAG was the dominant component of the total lipids, and it is the main form for ARA storage. The ARA enrichment stage occurred during 168–192h when the amount of total lipids maintained steady. Further analysis indicated that the newly formed ARA-TAG might come from the incorporation and modification of saturated and monounsaturated fatty acids in other lipid classes. This work could be helpful for further optimization of ARA-rich TAG production.

Dietary Enhancement of Selected Fatty Acid Biosynthesis in the Digestive Gland of Mytilus galloprovincialis Lmk.

The fatty acid composition of the digestive gland from the mussel Mytilus galloprovincialis subjected to three different dietary regimens for 30 days was analyzed. Samples were collected at the beginning and end of the trial to obtain a comprehensive picture of fatty acid dynamics. Group A was unfed; group B received a diet consisting of 100% Thalassiosira weissflogii and, thus, similar to natural food; and group C received a diet consisting of 100% wheat germ conferring a 18:2ω-6 abundance. Results indicate that fatty acid composition of lipid and phospholipid classes was affected by dietary treatments. However, adult mussel homeostatic skills minimized effects, and thus, only wheat germ diet deeply modified the fatty acid composition. Furthermore, in group C, the occurrence of the non-methylene-interrupted trienoic fatty acids was indicative of de novo fatty acid synthesis presumably because of active fatty acid elongation and Δ5 desaturation system, also supported by the general ω-3 polyunsaturated fatty acid decrease.

Mechanisms controlling lipid accumulation and polyunsaturated fatty acid synthesis in oleaginous fungi

AbstractPolyunsaturated fatty acids (PUFAs) are functional lipids that have been widely incorporated into several industrial sectors. Apart from animal- and plant-derived origins, oleaginous fungi belonging to Mucorales have been identified as promising alternatives for production of n-3 and n-6 PUFAs. It was found, in Mucorales fungi, that ATP:citrate lyase, acetyl-CoA carboxylase and malic enzyme trigger lipid overproduction, and biosynthesis of PUFA requires membrane-bound desaturases with fatty acyl substrate specificities. Accumulation of PUFAs in the cells is associated not only with the desaturation system, but it is also tightly bound with acyltransferases that facilitate the distribution of newly synthesized PUFA to individual lipid structures. Several physical parameters, such as temperature, aeration, and nutrient regimes, greatly affect either the lipid content or fatty acid composition among different Mucorales species. Conclusive evidence showed that the PUFA production yield of the fungi depends on the environmental control of “oleaginous” enzymes, and on the transcriptional expression of the desaturase genes. These valuable studies provide perspectives with biological rationale for microbial production of economically important lipids.

Chemiosmotic ATP synthesis in photosynthetically inactive chromoplasts from Narcissus pseudonarcissus L. linked to a redox pathway potentially also involved in carotene desaturation.

Mature chromoplasts from daffodil (Narcissus pseudonarcissus) flowers, although devoid of thylakoid structures, contain immunologically detectable alpha-subunits of ATP-synthase (H(+)-transporting ATP phosphohydrolase; EC 3.6.3.14). To show the presence of the entire functional protein complex, chromoplast membrane proteins were solubilized and reconstituted in phosphatidylcholine liposomes. The membranes were energized by an acid-base transition in the presence of a K(+)/valinomycin diffusion potential, and the initial rate of ATP synthesis was measured with a luciferin/luciferase assay. In addition, by demonstrating NADPH-dependent ATP synthesis, we show that an NAD(P)H-dependent respiratory redox pathway in chromoplasts, previously identified as an important constituent of the carotene desaturation system, proceeds concomitant with membrane energization.

Cloning, sequencing, and functional analysis of H-OLE1 gene encoding delta9-fatty acid desaturase in Hansenula polymorpha.

H-OLE1, a gene encoding delta9-fatty acid desaturase (FAD) in Hansenula polymorpha strain CBS 1976, was isolated by hybridization based upon its homology with the P-OLE1 gene cloned earlier from a related species, Pichia angusta IFO 1475. The sequence of the H-OLE1 gene revealed high structural conservation with delta9-FADs from various organisms. A putative 451-amino acid polypeptide encoded by the gene, like all other delta9-FADs, contained two domains: an N-terminal catalytic domain containing three conserved histidine clusters, and a C-terminal cytochrome b5-like domain which has been suggested to be involved in electron transport in desaturation reactions. The whole H-OLE1 gene complemented a H. polymorpha fad1 mutation leading to a defect in delta9-FAD. However, the unsaturated fatty acid requirement that the Saccharomyces cerevisiae ole1 mutant displays was complemented by only the open reading frame of H-OLE1 driven by S. cerevisiae glyceroaldehyde-3-phosphate dehydrogenase promoter, but not by the intact H-OLE1, suggesting that the H. polymorpha delta9-FAD was compatible with the desaturation system of S. cerevisiae whereas the promoter of the H-OLE1 gene had no activity in heterologous cells. It was shown by Northern hybridization that transcription of the H-OLE1 gene in H. polymorpha was slightly repressed by exogenous delta9-unsaturated fatty acid. An H. polymorpha disruption mutant (deltaH-OLE1) was created by transformation of an fad1/FAD1 diploid with disrupted H-OLE1::S-LEU2 linear DNA. It was shown by genetic and molecular analyses that input DNA was integrated in several copies into the chromosomal target to replace the mutated fad1 allele. Gas chromatography analysis showed identical fatty acid compositions in cells of both fad1 and deltaHOLE1 disruption mutants.

Transcriptional control of the low-temperature-inducible des gene, encoding the delta5 desaturase of Bacillus subtilis.

The Bacillus subtilis des gene encodes the cold-inducible Delta5 lipid desaturase involved in the formation of unsaturated fatty acids from saturated phospholipid precursors. Here, we describe the expression pattern of the des gene in response to a temperature downshift from 37 to 20 degrees C. We found that the synthesis of des mRNA is undetectable at 37 degrees C but dramatically induced upon the temperature downshift. Decay characteristics of the des transcript as well as the in vivo decay of B. subtilis bulk mRNA were investigated. The results showed that the stability of the des transcript as well as of bulk mRNA lasted substantially longer at 20 degrees C than at 37 degrees C. Functional expression of des at 37 degrees C was achieved by exchanging its promoter with the non-cold shock spac promoter. These data provide the first direct evidence that temperature-mediated control of transcription is the major mechanism regulating the mRNA levels of the B. subtilis desaturase. The present results also demonstrate that the only component of the desaturation system regulated by temperature is the desaturase enzyme.

Increase in hepatic content of oleic acid induced by dehydroepiandrosterone in the rat

The effects of dehydroepiandrosterone (DHEA) on the acyl composition of lipids in rat liver were studied. The content of oleic acid (18:1) in hepatic lipids was increased markedly by feeding rats a diet containing 0.5% (w/w) DHEA for 14 days. Treatment of rats with DHEA caused an increase in the activity of the terminal desaturase of the stearoyl-CoA desaturation system, without changing either the activity of NADH-cytochrome b 5 reductase or the microsomal content of cytochrome b 5. Among the changes observed in hepatic lipids, the increase in 18:1 content in phosphatidylcholine (PtdCho) was the most prominent; an approximately 2.5-fold increase in the proportion of 18:1 was induced at position 2, but not at position 1, by DHEA. This selective elevation of 18:1 at position 2 of PtdCho seems to be produced by the concerted actions of the induced 1-acylglycerophosphocholine (1-acyl-GPC) acyltransferase and the induced stearoyl-CoA desaturase. The content of 18:1 in serum lipids was unchanged by DHEA treatment, suggesting that secretion of lipids containing 18:1 into the circulation was not affected by DHEA. These results suggest that the elevation of hepatic content of 18:1 caused by DHEA treatment is mainly due to the induction of stearoyl-CoA desaturase.

Peroxisomal metabolism of adrenic acid; No Δ4 desaturase detected in rat liver peroxisomes

AbstractThe existence of a peroxisomal Δ4 desaturation of 22:4n‐6 and 22:5n‐3 to yield, respectively, 22:5n‐6 and 22:6n‐3 has been questioned. An alternative pathway has been formulated to include microsomal chain elongation and Δ6 desaturation and peroxisomal chain shortening. We incubated [1‐14C]adrenic acid (22:4n‐6) in a system for desaturation (i.e., in the presence of NADH) with purified rat liver peroxisomes. The fatty acids were separated as methyl derivatives by high‐performance liquid chromatography. Four ultraviolet‐absorbing product peaks appeared, three of which contained radioactivity, which we investigated as methyl, trimethylsilyl, and oxazoline derivatives on gas chromatography‐mass spectrometry. In addition to adrenic and arachidonic acids, the product peaks were trans‐enoyl, hydroxy, and keto derivatives of adrenic acid: the three first steps of β‐oxidation cycle. This indicated that the NAD‐dependent dehydrogenase step in the peroxisomal β‐oxidation cycle of adrenic acid was inhibited due to a high concentration of added NADH. Incubation in the presence of NAD instead of NADH reduced radioactivity in the peaks that corresponded to intermediates, while radioactivity in the acid‐soluble fraction increased considerably, consistent with a complete β‐oxidation cycle of adrenic to arachidonic acid. There were no indications of Δ4 desaturation in purified peroxisomes incubated in a standard desaturation system. Instead, adrenic acid as substrate underwent β‐oxidation.

Perfluorodecanoic acid enhances the formation of oleic acid in rat liver.

The feeding of perfluorodecanoic acid (PFDA) to male rats at a dietary concentration of 0.005% (w/w) for 7 days resulted in a marked increase in the activity of microsomal stearoyl-CoA desaturation in the liver. This increase in the overall desaturation activity was due to the induction of terminal desaturase among the components comprising the desaturation system. In contrast, PFDA inhibited desaturation in vitro, seemingly due to interference with electron transport through the desaturation system. Accordingly, PFDA can be an inducer and also an inhibitor of delta9-desaturation. PFDA feeding enhanced the conversion of radioactive stearic acid into oleic acid in the liver in vivo, indicating that the induction of delta9-desaturase by PFDA functions in vivo. PFDA feeding increased the mass of octadecenoic acid (C18:1) in the liver and the proportion of C18:1 in microsomal lipid. A highly significant linear correlation existed between the microsomal desaturase activity and the proportion of C18:1 in microsomal lipid when compared using rats in five different physiological states: control, PFDA-fed, p-chlorophenoxyisobutyric acid (clofibric acid)-fed, starved and starved/refed. These results suggest that the increase in the hepatic level of C18:1 caused by feeding of PFDA to rats can be explained by the common concept of regulation, i.e. the hepatic level of C18:1 is under the control of delta9-desaturase. The dietary administration of PFDA also increased the content of cytochrome P-450 and the activity of 7-ethoxycoumarin O-de-ethylase in the liver.

Purification and characterization of two forms of cytochrome b5 from an arachidonic acid-producing fungus, Mortierella hygrophila

Two forms of cytochrome b 5 have been purified from the microsomes of an arachidonic acid-producing fungus, Mortierella hygrophila IFO 5941, after detergent solubilization. They have monometic molecular masses of about 16 kDa and 19 kDa. Their absorption spectra are similar to those of mammalian cytochrome b 5s. Their amino acid compositions show some similarity to those of mammalian cytochrome b 5s, but the contents of some amino acids ( glycine, alanine, aspartic acid + asparagine, glutamic acid + glutamine, arginine, proline, histidine, leucine and lysine) are unique to the cytochrome b 5s of M. hygrophila. Some of their internal peptide sequences also show close homology with those of some mammals (approx. 65 to 67%), while some others show no or little homology. The addition of various acyl-CoAs to NADH-reduced microsomes caused an abrupt shiftdown of the steady state reduction level of cytochrome b 5. This indicates the increased utilization of electrons for the desaturation process and may suggest that the cytochrome b 5s of this fungus actually take part in its microsomel desaturation system for polyunsaturated fatty acid biosynthesis as electron carriers.

Plasma Lipid Changes in PRCD-affected and Normal Miniature Poodles Given Oral Supplements of Linseed Oil. Indications for the Involvement of N-3 Fatty Acids in Inherited Retinal Degenerations

It has previously been shown that miniature poodles with progressive rod-cone degeneration (PRCD) have lower plasma levels of docosahexaenoic acid (22:6n-3) than normal poodles and it has been suggested that affected animals have a defect in the metabolism of 22:6n-3. To test this hypothesis in vivo, PRCD-affected and normal miniature poodles were given daily oral supplements of linseed oil (enriched in 18:3n-3). Blood was drawn from food-deprived animals at predetermined times before, during and after supplementation, and plasma lipid fatty acids were analysed. There were no differences in the levels of 18:3n-3, 20:5n-3, and 22:5n-3 between affected and normal dogs. Therefore, there appears to be no abnormality in the elongation and desaturation system that takes 18:3n-3 to 22:5n-3. Surprisingly, the plasma level of 22:6n-3 was reduced in both groups following supplementation, but to a significantly greater extent in affected dogs. This resulted in a significantly higher 22:5n-3/22:6n-3 ratio in affected animals. These results support the earlier suggestion of an abnormality in 22:6n-3 metabolism in PRCD-affected miniature poodles. To determine the effect of n-3 supplementation on polyunsaturated fatty acid metabolism in dogs (not as a function of disease), results from both groups of dogs were pooled and compared at times before and near the end of supplementation. Dietary 18:3n-3 led to predictable increases in 18:3n-3, 20:5n-3, and 22:5n-3, but to a decrease in 22:6n-3. There was no effect of supplementation on 18:2n-6 and 22:5n-6 levels, although the levels of 20:4n-6 and 22:4n-6 were lowered. These results show that short-term supplementation of n-3 fatty acids affects both n-3 and n-6 fatty acid metabolism in dogs, as reflected in changes in plasma fatty acids.

Adrenic acid Δ4 desaturation and fatty acid composition in liver microsomes of spontaneously diabetic Wistar BB rats

We examined the Δ4 (n-6) desaturation and the fatty acid composition of liver microsomes in the insulin-dependent spontaneously diabetic Wistar Bio-Breeding (BB) rat. The desaturation of adrenic acid to n-6 docosapentaenoic acid was decreased in the normo- and hyperglycemic diabetic rats. Insulin treatment with 1.0 IU. 100 g body weight −1 twice a day for 2 days restored the reduced activity during the hypoglycemic period. The pattern of responses was similar to that of linoleic acid Δ6 and dihomo-γ-linolenic acid Δ5 desaturases, with a non-parallel relationship between the desaturation system and the glycemia. The microsomal fatty acid composition of BB rat liver reflected only partially to the Δ4 desaturation at different states of glycemia. Factors other than impaired desaturation system are involved in the fatty acid metabolism of spontaneously diabetic rats.

Effect of Vitamin B6Deficiency on Linoleic Acid Desaturation in the Arachidonic Acid Biosynthesis of Rat Liver Microsomes

The effect of vitamin B6 deficiency on the desaturation of linoleic acid (18:2n-6) to γ-linolenic acid (18:3n-6) in rat liver microsomes was studied. Rats fed with a vitamin B6-deficient 70% casein diet for 5 weeks displayed a striking decrease mainly in the activity of terminal Δ6-desaturase in the 18:2n-6 desaturation system, although the activity of the electron-transport enzyme, NADH-cytochrome b5 reductase, was also significantly decreased. The 18:2n-6 desaturation index, the arachidonic acid (20:4n-6)/ 18:2n-6 ratio in the liver microsomal total lipids and in their phosphatidylcholine (PC) fraction, decreased together with the microsomal Δ6-desaturase activity, these changes being quite faithfully reflected in the plasma lipid. Among the microsomal lipid components, the PC content significantly decreased. In addition, a highly positive correlation was found between the PC content and Δ6-desaturase activity in liver microsomes. In an in vitro experiment, the Δ6-desaturase activity decreased linearly with increasing rate of PC hydrolysis by phospholipase C. These results suggest that the altered PC content in microsomes induced by vitamin B6 deficiency in vivo may affect the 18:2n-6 desaturation and thus decrease the biosynthesis of 20:4n-6

Effect of n-6 and n-3 polyunsaturated fatty acids ingestion on rat liver membrane-associated enzymes and fluidity

The influences of diets having different fatty acid compositions on the fatty-acid content, desaturase activities, and membrane fluidity of rat liver microsomes have been analyzed. Weanling male rats (35–45 g) were fed a fat-free semisynthetic diet supplemented with 10% (by weight) marine fish oil (FO, 12.7% docosahexaenoic acid and 13.8% eicosapentaenoic acid), evening primrose oil (EPO, 7.8% γ-linolenic acid and 70.8% linoleic acid) or a mixture of 5% FO-5% EPO. After 12 weeks on the respective diets, animals fed higher proportions of (n-3) polyunsaturated fatty acids (FO group) consistently contained higher levels of 20:3(n-6), 20:5(n-3), 22:5(n-3), and 22:6(n-3), and lower levels of 18:2(n-6) and 20:4(n-6), than those of the EPO (a rich source of (n-6) polyunsaturated fatty acids) or the FO + EPO groups. Membrane fluidity, as estimated by the reciprocal of the order parameter S DPH, was higher in the FO than in the EPO or the FO + EPO groups, and the n-6 fatty-acid desaturation system was markedly affected.

Δ6 Desaturase Inhibition

The herbicide Norflurazone was shown to be an inhibitor of fatty acid of the Delta6 desaturation system. Treating cultures of the microalgae Spirulina platensis or Monodus subterraneus with this herbicide brought about a reduction in the level of gamma-linolenic acid and in an increase in the level of linoleic acid. In Monodus, the increase in linoleic acid made it more available for omega3 desaturation, resulting in an increase in the proportion of eicosapentaenoic acid. The proportion of arachidonic acid did not decrease albeit the drastic decrease in its precursor gamma-linolenic acid.

Age-related changes in lipid metabolism in rats: the consequence of moderate food restriction

Male Sprague-Dawley rats, 3 weeks of age, received one of the four following dietary manipulations until 9 months of age: group AD, fed ad libitum; group R, restricted to 75% of food intake of group AD; group RAD, restricted until 5 months of age and then fed ad libitum; group ADR, fed ad libitum until 5 months of age and then restricted. The concentration of serum total and HDL cholesterol tended to be higher in group AD than in all groups that had experienced food restriction. Liver cholesterol was higher in groups AD and RAD than in groups R and ADR. Activity of hepatic microsomal hydroxymethylglutaryl-CoA reductase was comparable, whereas sterol synthesis from mevalonate was significantly higher in groups R and ADR than in groups AD and RAD. Cholesterol 7α-hydroxylase activity also tended to be higher in groups R and ADR. It seems likely that hepatic cholesterol homeostasis functions effectively even when moderate food restriction was started after the growing period. In addition, food restriction reduced the ratio of arachidonate to linoleate, suggesting inhibition of the desaturation system.

The Effect of Dietary Essential Fatty Acid Deficiency on the Composition and Properties of the Liver Microsomal Membrane of Rats

The effect of dietary essential fatty acid (EFA) deficiency on lipid composition, fluidity and important enzyme and transport activities of liver microsomal membrane was studied in weanling rats. After 133 d of EFA deficiency, no difference was noticed in membrane phospholipid, cholesterol and protein levels, but a significant change occurred in the fatty acid composition of bilayer phospholipids. In EFA-deficient rats, linoleic (18:2(n-6] and arachidonic (20:4(n-6] acids were both severely lower while oleic (18:1(n-9], palmitoleic (16:1(n-7] and particularly 5,8,11-eicosatrienoic (20:3(n-9] acids were significantly higher than in controls. The higher level of the latter tended to compensate for the lower level of 20:4(n-6). Membrane fluidity, as estimated by the reciprocal of the order parameter S, was lower in the deficient rats than in the controls, and all the measured microsomal enzyme activities were markedly affected. NADH-Cyt b5 electron transferring system, coupled with the fatty acid desaturation system, was higher than in controls. In contrast, the cytochrome P450 complex activity was lower and some of the important liver detoxifying enzyme activities were lower due to physical-chemical changes in the microsomal membrane. Calcium uptake and Ca2+-ATPase activity were also significantly lower in EFA-deficient rats than in controls. It was concluded that fatty acid composition may be the major factor contributing to membrane fluidity and function and that EFA might play a role in regulating the intrinsic membrane protein activities.

Effects of long-term administration of clofibric acid on stearoyl-CoA desaturase, 1-acylglycerophosphorylcholine acyltransferase and fatty acyl composition of microsomal phosphatidylcholine in rat liver

Long-term effects of p-chlorophenoxyisobutyric acid (clofibric acid) on inductions of stearoyl-CoA desaturase and 1-acylglycerophosphorylcholine (1-acyl-GPC) acyltransferase, and on changes in fatty acyl composition of microsomal lipid in rat liver were studied. Male rats were fed clofibric acid at a dietary concentration of 0.25% for 2 or 22 weeks. Inductions of stearoyl-CoA desaturase and 1-acyl-GPC acyltransferase lasted throughout the long-term treatment and were the same as those of either young or aged rats which were treated with clofibric acid for 2 weeks. The long-term treatment of rats with clofibric acid scarcely affected components of stearoyl-CoA desaturation system other than terminal desaturase. In accordance with the induction of stearoyl-CoA desaturase, the increase in the proportion of octadecenoic acid in hepatic lipid lasted throughout the 22-week treatment. In the case of both of the long-term treatment and the short-term treatment of rats, the increase in the proportion of octadecenoic acid in microsomal phosphatidylcholine was due to the marked increase in the proportion of octadecenoic acid in position 2, but not position 1, of phosphatidylcholine. These changes in fatty acyl composition of phosphatidylcholine were not due to the alteration of the content of phosphatidylcholine in liver.

Isolation and partial characterization of a very long-chain fatty acid desaturation system from the cytosol of Mycobacterium smegmatis.

The supernatant fraction after 105,000 X g centrifugation of an extract of sonically disrupted cells of Mycobacterium smegmatis catalyzed the desaturation of lignoceroyl-CoA to a delta15-monounsaturated derivative in the presence of molecular oxygen and NADPH. This desaturation system was separated by ammonium sulfate fractionation, gel filtration, DEAE-cellulose column chromatographies, and affinity column chromatography on immobilized dye, into three components; a NADPH-oxidase, a ferredoxin-containing fraction and a desaturase, all of which were required for the reconstituted desaturation system for lignoceroyl-CoA. This system was inhibited by FMN and ferrous ions but not by KCN. All of these features clearly distinguish this system from the previously known fatty acid desaturation systems of various origins.