Numerous Δ12-, Δ15- and multifunctional membrane fatty acid desaturases (FADs) have been identified in fungi, revealing great variability in the enzymatic specificities of FADs involved in biosynthesis of polyunsaturated fatty acids (PUFAs). Here, we report gene isolation and characterization of novel Δ12/Δ15- and Δ15-FADs named CpFad2 and CpFad3, respectively, from the opportunistic pathogenic yeast Candida parapsilosis. Overexpression of CpFad3 in Saccharomyces cerevisiae strains supplemented with linoleic acid (Δ9,Δ12-18:2) and hexadecadienoic acid (Δ9,Δ12-16:2) leads to accumulation of Δ15-PUFAs, i.e., α-linolenic acid (Δ9,Δ12,Δ15-18:3) and hexadecatrienoic acid with an unusual terminal double bond (Δ9,Δ12,Δ15-16:3). CpFad2 produces a range of Δ12- and Δ15-PUFAs. The major products of CpFad2 are linoleic and hexadecadienoic acid (Δ9,Δ12-16:2), accompanied by α-linolenic acid and hexadecatrienoic acid (Δ9,Δ12,Δ15-16:3). Using GC/MS analysis of trimethylsilyl derivatives, we identified ricinoleic acid (12-hydroxy-9-octadecenoic acid) as an additional product of CpFad2. These results demonstrate that CpFAD2 is a multifunctional FAD and indicate that detailed analysis of fatty acid derivatives might uncover a range of enzymatic selectivities in other Δ12-FADs from budding yeasts (Ascomycota: Saccharomycotina).

• MeSH
• Candida metabolism   MeSH
• Fatty Acid Desaturases metabolism   MeSH
• Phylogeny MeSH
• Linoleic Acid metabolism   MeSH
• Ricinoleic Acids metabolism   MeSH
• Molecular Sequence Data MeSH
• Fatty Acids, Unsaturated metabolism   MeSH
• Saccharomyces cerevisiae metabolism   MeSH
• Amino Acid Sequence MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

Bumblebee males (Hymenoptera) produce species-specific labial gland secretions called marking pheromones (MPs). MPs generally consist of terpenoids and fatty-acid-derived aliphatic compounds with various chain lengths predominantly containing one or no double bonds. The unsaturated fatty-acid-derived MP components were hypothesized to be produced by fatty acid desaturases (FADs) that exhibit diverse substrate specificities. To address this hypothesis, we isolated and functionally characterized FADs from three bumblebee species: Bombus lucorum, Bombus terrestris, and Bombus lapidarius. By employing RNA sequencing of the male labial glands and fat bodies of B. lucorum and B. terrestris, we identified five paralogous FAD-like sequences but only two FAD lineages were abundant and differentially expressed in the labial glands. We found that abundant FAD lineages were also expressed in the labial gland and fat body of Bombus lapidarius. Functional characterization of FADs in a yeast expression system confirmed that Δ4-FADs exhibited a unique Δ4-desaturase activity exclusively on 14-carbon fatty acyls and Δ9-FADs displayed Δ9-desaturase activity on 14- to 18-carbon fatty acyls. These results indicate that Δ9-FADs are involved in the biosynthesis of major unsaturated components of MPs in B. lucorum and B. lapidarius despite the diverse MP composition of these bumblebee species. The contribution of lipases, acyltransferases, esterases, and fatty acid reductases to production of the species-specific MP composition is also discussed in light of the transcriptomic data obtained in this study.

• MeSH
• Fatty Acid Desaturases isolation & purification   metabolism   MeSH
• Pheromones biosynthesis   MeSH
• Phylogeny MeSH
• Insect Proteins isolation & purification   metabolism   MeSH
• Sequence Analysis, DNA MeSH
• Stearoyl-CoA Desaturase metabolism   MeSH
• Bees enzymology   genetics   MeSH
• Animals MeSH
• Check Tag
• Male MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

Východisko. Složení esterifikovaných mastných kyselin v plazmě je u metabolického syndromu a dalších stavů inzulínové rezistence významně změněno. Profil mastných kyselin v plazmě je dán složením tuku přijímaného potravou a metabolickými pochody přeměny mastných kyselin, tedy de novo lipogenezí, β-oxidací i přeměnou v důsledku oxidačního stresu. Cílem této práce bylo zjistit složení mastných kyselin v hlavních lipidových třídách plazmy ve vztahu k inzulínové rezistenci, k některým polymorfizmům kandidátních genů, jejichž aktivita s inzulínovou rezistencí souvisí, a ke složení lipoproteinů i parametrům lipoperoxidace. Metody a výsledky. Do studie bylo zařazeno 95 osob s metabolickým syndromem (56 M/39 F) a 195 zdravých osob (99 M/96 F). Vyšetřili jsme základní klinická data, parametry glukózové homeostázy, koncentrace lipidů v plazmě a konjugované dieny v LDL. Mastné kyseliny byly stanoveny kapilární plynovou chromatografií. Polymorfizmy apolipoproteinu E, střevní izoformy proteinu vázajícího mastné kyseliny (Ala54Thr) a γ-2 izoformy receptoru aktivovaného peroxizomálními proliferátory (Ala12Pro) byly analyzovány kombinací metod polymerázové řetězové reakce a určování polymorfizmů délky restrikčních fragmentů. Osoby s metabolickým syndromem měly vyšší koncentrace CRP a konjugovaných dienů v LDL. Ve všech lipidových třídách jsme prokázali pokles celkové koncentrace vícenenasycených mastných kyselin řady n-6 a vzestup koncentrace nasycených mastných kyselin. Z jednotlivých kyselin byl významný pokles koncentrace kyseliny linolové a vzestup obsahu kyseliny palmitové a palmitolejové. Z výsledků lze usuzovat na vzestup aktivit Δ9 desaturázy kyseliny palmitové, Δ6 desaturázy a elongázy kyseliny linolové. Koncentrace konjugovaných dienů v LDL negativně korelovaly s kyselinou linolovou. Nezjistili jsme vazbu klinických ani laboratorních parametrů na homozygocii vyšetřených polymorfizmů. Závěry. U metabolického syndromu jsou změny profilu mastných kyselin důsledkem zvýšené lipogeneze a akcentovaného oxidačního stresu.

Background. Composition of the nonesterified fatty acids in plasma in metabolic syndrome patients and in other syndromes of insulin resistance is altered. Fatty acid profile in plasma is related to the composition of dietary fat and to the metabolic changes of fatty acids, e.g. to de novo lipogenesis, β-oxidation and conversion accompanying the oxidative stress. The aim of the work was to study the fatty acid composition in the major plasma lipid classes in relation to the insulin resistance, to some polymorphisms of candidate genes with activity related to insulin resistance, and to the lipoprotein composition and parameters of lipid peroxidation. Methods and Results. 95 patients with metabolic syndrome (56 M/39 F) and 195 healthy persons (99 M/96 F) were included into the cohort. Basic clinical data, parameters of glucose homeostasis, lipid concentration in plasma and conjugated diens in LDL were determined. Fatty acids were detected by capillary gas chromatography. Polymorphisms of apolipoprotein E, intestinal isoforms of fatty acid binding protein (Ala54Thr) and γ-2 isoforms of peroxisomal activated receptor (Ala12Pro) were analyzed using combination of polymerase chain reaction methods and by the detection of polymorphisms of the restriction fragment length. Persons with metabolic syndrome had higher concentrations of CRP and conjugated diens in LDL. In all lipid classes we proved a decreased concentration of n-6 polyunsaturated fatty acids and an increase of unsaturated fatty acids. From all the acids, the only significant was the decrease of linolic acid concentration and the increase of palmitic and palmitoyl acids. Results showed an increase of Δ9 palmitic acid desaturase activity, Δ6 linolic acid desaturase and elongase activity. Concentration of conjugated diens in LDL inversely correlated with linolic acid. Clinical or laboratory parameters and homozygotic combination of polymorphism studied were not mutually related. Conclusions. Changes in the profile of fatty acids during the metabolic syndrome results from the elevated lipogenesis and from the higher level of oxidative stress.

• MeSH
• Fatty Acid Desaturases genetics   metabolism   MeSH
• Research Support as Topic MeSH
• Humans MeSH
• Fatty Acids analysis   metabolism   MeSH
• Metabolic Syndrome metabolism   MeSH
• Oxidative Stress radiation effects   MeSH
• Lipid Peroxidation genetics   immunology   MeSH
• Polymorphism, Genetic genetics   immunology   MeSH
• Check Tag
• Humans MeSH

Arbuscular mycorrhizas (AMs) have the ability to enhance drought tolerance of citrus, but the underlying mechanisms have not been clearly elucidated. Considering the strong association of cell membrane fatty acid (FA) unsaturation with plant drought tolerance, the present study hypothesized that AM fungi (AMF) modulated the composition and unsaturation of FAs to enhance drought tolerance of host plants. Drought-sensitive citrus rootstocks, trifoliate orange (Poncirus trifoliata) seedlings, were inoculated with AMF (Funneliformis mosseae) for 3 months and were subsequently exposed to drought stress (DS) for 8 weeks. Mycorrhizal seedlings exhibited better plant growth performance, higher leaf water potential and lower root abscisic acid concentrations under both well-watered (WW) and DS conditions. Arbuscular mycorrhiza fungus inoculation considerably increased root methyl oleate (C18:1), methyl linoleate (C18:2) and methyl linolenate (C18:3N3) concentrations under both WW and DS conditions, and root methyl palmitoleate (C16:1) concentrations under WW, while it decreased root methyl stearate (C18:0) levels under both WW and DS. These changes in the composition of FAs of mycorrhized roots resulted in higher unsaturation index of root FAs, which later aided in reducing the oxidative damage on account of lower concentration of malondialdehyde and superoxide radicals. The changes of these FAs were a result of AMF-up-regulating root FA desaturase 2 (PtFAD2), FA desaturase 6 (PtFAD6) and Δ9 FA desaturase (PtΔ9) genes under WW and PtFAD2, PtFAD6 and Δ15 FA desaturase (PtΔ15) genes under DS conditions. Our results confirmed that mycorrhization brought significant changes in root FA compositions, in addition to regulation of gene expression responsible for increasing the unsaturation level of FAs, a predisposing physiological event for better drought tolerance of citrus.

• MeSH
• Citrus * MeSH
• Glomeromycota * MeSH
• Plant Roots MeSH
• Mycorrhizae * MeSH
• Droughts MeSH
• Poncirus * MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

Pancreatic cancer (PC) ranks as the fourth cause of cancer-related deaths in the Czech Republic. Evidence exists that deregulation of fatty acid (FA) metabolism is connected with some malignancies; therefore, we decided to analyze FA profile in plasma lipid classes in patients with PC with relation to tumor staging, nutritional status, and survival. The study included 84 patients (47 males, 37 females) with PC and 68 controls (36 males, 32 females). FA patterns were analyzed in plasma lipid classes by gas-chromatography. We observed increased proportion of total monounsaturated FA (MUFA) in PC group in all plasma lipid classes. These changes were connected with increased Δ9-desaturase (SCD1) and Δ5-desaturase indices. Correlations of dihomo-γ-linolenic acid (DHGLA) with these variables were opposite. Longer survival of patients was connected with higher content of EPA, DHA, and with lower SCD1 index, respectively. Plasma phospholipid proportions of α-linolenic acid, DHGLA, EPA, and n-3 polyunsaturated fatty acids displayed negative trend with tumor staging. Plasma lipid FA pattern in PC patients resulted from decreased dietary fat intake and increased de novo synthesis of FA with transformation into MUFA. Changes in FA profile implicated some pathophysiological mechanisms responsible for disturbed FA metabolism in PC and importance of appropriate nutritional support.

• MeSH
• Cholesterol blood   MeSH
• Phospholipids blood   MeSH
• Incidence MeSH
• 8,11,14-Eicosatrienoic Acid blood   MeSH
• alpha-Linolenic Acid blood   MeSH
• Docosahexaenoic Acids blood   MeSH
• Middle Aged MeSH
• Humans MeSH
• Fatty Acids blood   MeSH
• Lipid Metabolism MeSH
• Pancreatic Neoplasms epidemiology   MeSH
• Nutritional Status MeSH
• Malnutrition epidemiology   MeSH
• Aged MeSH
• Case-Control Studies MeSH
• Triglycerides blood   MeSH
• Check Tag
• Middle Aged MeSH
• Humans MeSH
• Male MeSH
• Aged MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Geographicals
• Czech Republic MeSH

A group of 240-day-old Ross cockerels were used in a 4-week experiment to assess the effect of the fat type on the intestinal digestibility of fatty acids (FAs), the FA profiles of breast meat and abdominal fat, and the mRNA expression of six hepatic lipid-related genes. Experimental diets were supplemented with rapeseed oil, pork lard or palm oil at 60 g/kg. In the control diet, wheat starch was substituted for the fat source. The highest ileal digestibility of the fat and all FAs (except stearic acid) was observed in chickens fed lard. The content of fat in the breast meat of chickens was not significantly influenced by the fat supplements. The FA profiles of breast meat and abdominal fat reflected the FA composition of the diet. In the meat of chickens fed rapeseed oil, oleic acid was the predominant FA. Palmitic acid was the most abundant FA in the meat of chickens fed lard or palm oil. Oleic acid was the most abundant FA in the abdominal fat of all chickens. The highest mRNA expression of desaturases (Δ5-, Δ6- and Δ9-) was observed in chickens fed palm oil. The mRNA expression of hepatic FA synthase was higher in chickens fed palm oil or lard than in chickens fed rapeseed oil. The expression of HMG-CoA reductase was higher in chickens fed palm oil than in those fed rapeseed oil or lard. It can be concluded that rapeseed oil and lard are better sources of lipids than palm oil. These former two sources contain more digestible fatty acids and provide a lower concentration of SFAs in the meat and fat of chickens.

• MeSH
• Dietary Fats metabolism   MeSH
• Gene Expression * MeSH
• Animal Feed MeSH
• Chickens genetics   metabolism   MeSH
• Fatty Acids metabolism   MeSH
• RNA, Messenger genetics   metabolism   MeSH
• Lipid Metabolism * genetics   MeSH
• Organ Specificity MeSH
• Intestines metabolism   MeSH
• Digestion * MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH