Fatty Acid Desaturase Gene Research Articles

Quantitative Proteomic Analysis of Low Linolenic Acid Transgenic Soybean Reveals Perturbations of Fatty Acid Metabolic Pathways.

To understand the effect of fatty acid desaturase gene (GmFAD3) silencing on perturbation of fatty acid (FA) metabolic pathways, the changes are compared in protein profiling in control and low linolenic acid transgenic soybeans using tandem mass tag based mass spectrometry. Protein profiling of the transgenic line unveiled changes in several key enzymes of FA metabolism. This includes enzymes of lower abundance; fabH, fabF, and thioestrase associated with FA initiation, elongation, and desaturation processes and LOX1_5, ACOX, ACAA1, MFP2 associated with β-oxidation of α-linolenic acids pathways. In addition, the GmFAD3 silencing results in a significant reduction in one of the major allergens, Gly m 4 (C6T3L5). These results are important for exploring how plants adjust in their biological processes when certain changes are induced in the genetic makeup. A complete understanding of these processes will aid researchers to alter genes for developing value-added soybeans.

Genome-wide identification, characterization and in-silico profiling of genes encoding FAD (fatty acid desaturase) proteins in chickpea (Cicer arietinum L.)

Fatty acid desaturases (FADs) are involved in introducing double bonds into the hydrocarbon chains of fatty acids leading to the formation of unsaturated fatty acids. These unsaturated fatty acids are known to play important role in plant development and generating stress tolerance against various environmental stresses. In this study, 21 full-length FAD genes were identified in Desi chickpea (Cicer arietinum L.) and 18 FAD genes in Kabuli chickpea genome by a thorough annotation exercise. An extensive analysis was done to establish their chromosomal locations, construct a phylogeny, structure prediction, motif analysis, in-silico expression analysis and co-expression with other interconnected pathways. In desi chickpea, out of 21 FAD genes, 19 were located on seven chromosomes and two additional scaffolds whereas in Kabuli, out of 18 FAD genes, 17 were located on seven chromosomes and an additional scaffold. Identified soluble and membrane-bound CaFADs were phylogenetically clustered into four and eight subfamilies, respectively with their counterparts from other plants. It was observed that gene structures, motif composition and positions were highly conserved in each subfamily. Furthermore, in-silico gene expression analysis of CaFAD genes in various tissues as well as under abiotic stress conditions revealed differential expression patterns in different tissues and developmental stages. Nine FAD genes were down-regulated in all three abiotic stresses as compared to control. Whereas, five FAD genes were up-regulated in shoot tissue during drought, two FAD genes during salinity and three FAD genes were up-regulated during cold stress. In root tissue, 9 FAD genes were down-regulated and 3 FAD genes were up-regulated during all three abiotic stresses. In co-expression analysis, fatty acid synthesis, TAG synthesis genes and Jasmonic acid (JA) biosynthesis genes were expressed more during flower development and shoot apical meristem. JA signaling gene (CaJAR1) was also expressed more in bud, shoot apical meristem and seeds. No other relationship was observed with respect to modulation during abiotic stresses.

The Effect of an Infant Formula Supplemented with AA and DHA on Fatty Acid Levels of Infants with Different FADS Genotypes: The COGNIS Study.

Polymorphisms in the fatty acid desaturase (FADS) genes influence the arachidonic (AA) and docosahexaenoic (DHA) acid concentrations (crucial in early life). Infants with specific genotypes may require different amounts of these fatty acids (FAs) to maintain an adequate status. The aim of this study was to determine the effect of an infant formula supplemented with AA and DHA on FAs of infants with different FADS genotypes. In total, 176 infants from the COGNIS study were randomly allocated to the Standard Formula (SF; n = 61) or the Experimental Formula (EF; n = 70) group, the latter supplemented with AA and DHA. Breastfed infants were added as a reference group (BF; n = 45). FAs and FADS polymorphisms were analyzed from cheek cells collected at 3 months of age. FADS minor allele carriership in formula fed infants, especially those supplemented, was associated with a declined desaturase activity and lower AA and DHA levels. Breastfed infants were not affected, possibly to the high content of AA and DHA in breast milk. The supplementation increased AA and DHA levels, but mostly in major allele carriers. In conclusion, infant FADS genotype could contribute to narrow the gap of AA and DHA concentrations between breastfed and formula fed infants.

Seed development and nutrient accumulation as affected by light shading in oilseed peony (Paeonia ostii ‘Feng Dan’)

Seed yield and nutritional quality are dependent on seed development in oil crops. In a previous study, we showed that light shading improved seed yield and quality in oilseed peony, but the underlying mechanisms driving light shading impacts on seed yield and nutritional quality as mediated through seed development remain unclear. To develop an understanding of these mechanisms, oilseed peony trees were shaded by different density polyethylene nets starting at 28 days after pollination (DAP) and continuing until harvest. The seed volume, weight, and nutritional quality parameters, as well as the expression of genes related to the synthesis of unsaturated fatty acids (UFAs), were examined at 14-day intervals. Results showed that hundred-seed volume (Vh), fresh weight (Wf), and dry weight (Wd) increased by 8.2, 15.2, and 33.0 times, respectively, from 14 to 70 DAP. During the same period, the contents of starch, soluble protein, crude oil, and UFAs increased by 2.8, 3.7, 20.4, and 21.1 times, respectively. From 71 to 98 DAP, Vh, Wf, and Wd increased by 4.1, 5.4, and 20.8%, respectively. During this period, the contents of soluble protein, crude oil, and UFAs increased by 36.8, 51.4, and 49.5%, respectively. From 99 to 112 DAP, Vh, Wf, and Wd slightly increased, and the content of soluble protein increased by 25.2%. In contrast, the contents of soluble sugar, starch, and crude oil decreased by 28.3, 15.1, and 6.3%, respectively. Thus, seed development of oilseed peony can be divided into three periods of rapid growth (before 70 DAP), slow growth (71–98 DAP), and maturation (99–112 DAP). At 98 DAP, light shading increased the content of soluble sugar by 8.3%. At 112 DAP, light shading increased Vh, Wf, Wd, the content of crude oil, and UFAs by 6.9, 8.3, 8.3, 6.6, and 9.6%, respectively. The expression level of FAD2 in peony seed was up-regulated by 87.1 and 219.3% at 98 and 112 DAP, respectively. The expression level of FAD8 was up-regulated by 10.8, 71.7, 162.1, and 371.3% at 70, 84, 98, and 112 DAP, respectively. The expression level of SAD was up-regulated by 53.5 and 150.3% at 98 and 112 DAP, respectively. It is thus concluded that light shading significantly improved seed yield and nutritional quality parameters via increased seed growth and the expression of fatty acid desaturase genes during the slow growth and the maturation periods of seed development.

Reduced delta-6 desaturase activity partially protects against high-fat diet-induced impairment in whole-body glucose tolerance

Delta-6 desaturase (D6D), which is encoded by the fatty acid desaturase (Fads2) gene, is the rate-limiting enzyme for the endogenous production of n-3 long-chain polyunsaturated fatty acids. The absence of D6D activity in Fads2−/− knockout mice results in the inability to produce eicosapentaenoic acid and docosahexaenoic acid, and has previously been associated with altered glucose and lipid metabolism. Skeletal muscle is a major site for insulin-stimulated glucose disposal; however, the consequences of reduced D6D activity on skeletal muscle metabolism are unknown. The objective of this study was to examine the role of a partial reduction in D6D activity on whole-body glucose tolerance, skeletal muscle fatty acid profiles and protein content of key markers of carbohydrate and fat signaling pathways in the context of both low- and high-fat diets. Male C57BL/6J heterozygous (Fads2+/−) and wild-type (WT) mice were fed either a low-fat (16% kcal from fat) or high-fat (HFD; 45% kcal from fat) diet for 21 weeks. Fads2+/− mice were protected from the HFD-induced impairment in glucose tolerance. Unexpectedly, HFD-fed Fads2+/− mice had reduced GLUT4 skeletal muscle protein content compared to their WT counterparts. No changes were detected in total protein content of key markers of fatty acid uptake, glycogen formation or substrate oxidation. This study shows that reduced D6D activity is partially protective against HFD-induced impairments in whole-body glucose tolerance but does not appear to be due to increased muscle GLUT4 content or total content of proteins regulating substrate utilization.

The Evolution and Biocatalysis of FAD2 Indicate Its Correlation to the Content of Seed Oil in Plants.

Unsaturated fatty acids are the main components of vegetable oils. Fatty acid desaturase 2 (FAD2) catalyzes oleic acid (OA) into linoleic acid (LA) transformations, which are essential to the profile of FAs in seeds. To further understand the roles of FAD2s in the synthesis of oil, the evolution and biocatalysis of FAD2s were comprehensively analyzed. The evolution history of the FAD2 gene family showed that most of the FAD2 genes formed monophyletic clades except in eudicots. The FAD2 genes in some eudicots diverged into constitutive and seed-specific expression clades. Notably, the biocatalysis of seed-specific or -abundant expression FAD2s in soybean, perilla, rice, and spruce revealed that their catalytic activity was strongly correlated with the total oil content of their seeds in nature. Additionally, it was found that I and Y in site 143 of GmaFAD2-1 were strictly conserved in the seed-specific and constitutive expression clades of Fabaceae, respectively. Furthermore, the site-directed mutation demonstrated that I and Y are vital to improving and reducing the activity of GmaFAD2s. Therefore, the results indicate that the activity of FAD2s in seeds might be a reference to the total oil content of seeds, and site 143 might have been specifically evolved to be required for the activity of FAD2s in some expression-diverged eudicots, especially in legumes.

Characteristics of membrane-bound fatty acid desaturase (FAD) genes in Brassica napus L. and their expressions under different cadmium and salinity stresses

The membrane-bound fatty acid desaturases (FADs) play vital roles in regulating plant fatty acid (FA) compositions which directly affect oil quality in rapeseed (Brassica napus L.). In the present study, a total of 68 putative FAD genes were identified in the crop and were phylogenetically clustered into four subfamilies. Genomic localization revealed that the genes were distributed across 21 chromosomes at various densities. The structures of paralogous genes were considerably conserved in the number of exons and in the length of introns. It was found that the expansion of the B. napus FAD family was mainly attributable to whole genome duplication and segmental duplication. Rapeseed seedlings were grown hydroponically under three levels (0, 250, and 500 μM) of cadmium (Cd) and three levels (0, 50, and 100 mM) of salinity. After one week of treatment, the roots and leaves of seedlings were harvested to study the expression patterns of 68 selected BnFADs in response to the different stresses. The results from qRT-PCR analyses suggested that the FAD genes in rapeseed were differentially expressed under various abiotic stresses. The transcript levels of the FAD genes ADS4.8, ADS4.9, ADS4.10, ADS9.1 and SLD8 in the leaves of Cd-tolerant cultivar ZD619 were significantly higher than those in the leaves of sensitive cultivar ZD622 under the 250 μM Cd stress condition. Moreover, the expression levels of 8FAD genes (ADS4.1, ADS4.4, ADS4.9, ADS9.1, ADS9.2, SLD8, FAD7.4 and FAD7.5) in ZD619 roots were obviously enhanced with the increase of salinity concentrations. These results indicate the likelihood of FAD genes involved in Cd and salinity tolerance. This study provides insight on membrane-bound FAD genes involvement in tolerance to abiotic stresses and would provide some more candidate genes for the crop improvement.

Double bonds of unsaturated fatty acids differentially regulate mitochondrial cardiolipin remodeling

BackgroundSupplemented fatty acids can incorporate into cardiolipin (CL) and affect its remodeling. The change in CL species may alter the mitochondrial membrane composition, potentially disturbing the mitochondrial structure and function during inflammation.MethodTo investigate the effect of the unsaturation of fatty acids on CL, we supplemented macrophage-like RAW264.7 cells with 18-carbon unsaturated fatty acids including oleic acid (OA, 18:1), linoleic acid (LA, 18:2), α-linolenic acid (ALA, 18:3), γ-linolenic acid (GLA, 18:3), and stearidonic acid (SDA, 18:4). Mitochondrial changes in CL were measured through mass spectrometry.ResultOur data indicated that OA(18:1) was the most efficient fatty acid that incorporated into CL, forming symmetrical CL without fatty acid elongation and desaturation. In addition, LA(18:2) and ALA(18:3) were further elongated before incorporation, significantly increasing the number of double bonds and the chain length of CL. GLA and SDA were not optimal substrates for remodeling enzymes. The findings of RT-qPCR experiments revealed that none of these changes in CL occurred through the regulation of CL remodeling- or synthesis-related genes. The fatty acid desaturase and transportation genes—Fads2 and Cpt1a, respectively—were differentially regulated by the supplementation of five unsaturated 18-carbon fatty acids.ConclusionsThe process of fatty acid incorporation to CL was regulated by the fatty acid desaturation and transportation into mitochondria in macrophage. The double bonds of fatty acids significantly affect the incorporation process and preference. Intact OA(18:1) was incorporated to CL; LA(18:2) and ALA(18:3) were desaturated and elongated to long chain fatty acid before the incorporation; GLA(18:3) and SDA(18:4) were unfavorable for the CL incorporation.

Molecular cloning and functional characterization of elongase (elovl5) and fatty acyl desaturase (fads2) in sciaenid, Nibea diacanthus (Lacepède, 1802)

In the present paper, we investigated the molecular cloning and functional characterization of elongase of very long chain fatty acid (elovl) and fatty acyl desaturase (fads) genes in a marine teleost, Nibea diacanthus. The elongase cDNA sequence encoded a polypeptide of 294 amino acids exhibiting Elovl5 activity, which effectively elongated both C18 (18:2n−6, 18:3n−3 and 18:3n−6) and C20 (20:4n−6 and 20:5n−3) polyunsaturated fatty acids. The desaturase cDNA sequence specified a polypeptide of 445 amino acids indicating Δ6 desaturation activity, which coul converted C18:2n−6 and C18:3n−3 to C18:3n−6 and C18:4n−3, respectively. Tissue distribution analysis by quantitative real-time PCR revealed that the elovl5 was primarily expressed in intestine and liver, while the fads2 was mainly expressed in liver and brain. These results increase our knowledge of the ability of endogenous highly unsaturated fatty acids (HUFA) biosynthesis in marine carnivorous fish.

Identification and characterization of a delta-12 fatty acid desaturase gene from marine microalgae Isochrysis galbana

The cDNA of the delta-12 fatty acid desaturase gene, IgFAD2, was cloned from the marine microalgae Isochrysis galbana, a species capable of producing docosahexaenoic acid. Sequence analysis indicated that the open reading frame measured a length of 1 158 bp and encoded 386 amino acids with a predicted molecular weight of 42.8 kDa and an isoelectric point of 9.2. Computational analysis of the protein sequence of IgFAD2 showed typical features of membrane-bound desaturase such as three conserved histidine boxes along with four membrane-spanning regions that were universally present among plant desaturases. Quantitative real-time PCR results showed that the abundance of IgFAD2 transcript was significantly upregulated under different environmental stresses including low temperature (15°C), high salinity (salinity of 62 and 93), and nitrogen starvation (220 μmol/L). Heterologous expression indicated that yeast cells transformed with a plasmid construct containing IgFAD2 could convert endogenous oleic acid (18:1Δ9, OA) into linoleic acid (18:2Δ9, 12, LA). These findings confirm that I. galbana IgFAD2 plays important roles in the biosynthetic pathways of unsaturated fatty acids.

Genome-wide identification and expression of eight fatty acid desaturase genes, and the fatty acid profile, in the marine rotifer Brachionus koreanus fed the alga Tetraselmis suecica

Fatty acid desaturases catalyze the formation of a C=C double bond from a C–C single bond in fatty acids, leading to diversification of the fatty acid pool. In this study, we identified the fatty acid desaturase genes in the monogonont marine rotifer Brachionus koreanus, measured the messenger RNA (mRNA) expression, and quantified fatty acid composition under feeding with the alga Tetraselmis suecica. Eight fatty acid desaturase genes were identified and classified by phylogenetic analysis. Fatty acid desaturases in the rotifer B. koreanus were categorized into two Δ4 desaturases, five Δ5/6 desaturases, and one Δ9 desaturase. mRNA expression of B. koreanus under the alga T. suecica-fed condition clearly indicated that transcriptional levels of desaturase genes were increased compared to the non-T. suecica-fed group. Also, the distribution of fatty acids of B. koreanus fed T. suecica was more balanced compared to that of T. suecica. These results provide a better understanding of the role of fatty acid desaturases in B. koreanus fed the alga T. suecica.

Inflammatory response to dietary linoleic acid depends on FADS1 genotype

The health benefits of substituting dietary polyunsaturated fatty acids (PUFAs) for saturated fatty acids are well known. However, limited information exists on how the response to dietary intake of linoleic acid (LA; 18:2n-6) is modified by polymorphisms in the fatty acid desaturase (FADS) gene cluster. The aim of the current study was to test the hypothesis that the FADS1 rs174550 genotype modifies the effect of dietary LA intake on the fatty acid composition of plasma lipids, fasting glucose, and high-sensitivity C-reactive protein (hsCRP). Associations were investigated between genotype, plasma PUFAs, fasting glucose, and hsCRP concentrations in the cross-sectional, population-based Metabolic Syndrome in Men cohort (n=1337). In addition, 62 healthy men from the cohort who were homozygotes for the TT or CC genotype of the FADS1 rs174550 were recruited to a 4-wk intervention (FADSDIET) with an LA-enriched diet. The fatty acid composition of plasma PUFAs and concentrations of plasma fasting glucose, serum hsCRP, and plasma lipid mediators (eicosanoids and related analogs) were measured at the beginning and end of the 4-wk intervention period. In the FADSDIET trial, the plasma LA proportion increased in both genotype groups in response to an LA-enriched diet. Responses in concentrations of serum hsCRP and plasma fasting glucose and the proportion of arachidonic acid (20:4n-6) in plasma phospholipids and cholesteryl esters differed between genotype groups (interaction of diet × genotype, P<0.05). In TT homozygous subjects, plasma eicosanoid concentrations correlated with the arachidonic acid proportion in plasma and with hsCRP (r=0.4-0.7, P<0.05), whereas in the CC genotype there were no correlations. Our findings show that the FADS1 genotype modifies metabolic responses to dietary LA. The emerging concept that personalized dietary counseling should be modified by the FADS1 genotype needs to be tested in larger randomized trials. The study was registered at clinicaltrials.gov as NCT02543216.

Δ12 fatty acid desaturase gene from Geotrichumcandidum in cheese: molecular cloning and functional characterization.

Soft cheese with white rind lacks essential fatty acids (EFAs), and as a result its long‐term consumption may lead to various kinds of cardiovascular and cerebrovascular diseases, such as hyperlipidemia, hypertension, and atherosclerosis. Geotrichum candidum is a dimorphic yeast that plays an important role in the ripening of mold cheese. A gene coding for Δ12 fatty acid desaturase, a critical bifunctional enzyme desaturating oleic acid (OA) and linoleic acid (LA) to produce LA and α‐linolenic acid (ALA), respectively, was isolated from G. candidum, and then cloned and heterologously expressed in Saccharomyces cerevisiae. This gene, named GcFADS12, had an open reading frame of 1257 bp and codes for a protein of 419 amino acids with a predicted molecular mass of 47.5 kDa. Characterization showed that GcFADS12 had the ability to convert OA to LA and LA to ALA, and the conversion rates for OA and LA were 20.40 ± 0.66% and 6.40 ± 0.57%, respectively. We also found that the protein product of GcFADS12 catalyzes the conversion of the intermediate product (LA) to ALA by addition of OA as the sole substrate. The catalytic activity of GcFADS12 on OA and LA was unaffected by fatty acid concentrations. Kinetic analysis revealed that GcFADS12 had stronger affinity for the OA than for the LA substrate. This study offers a solid basis for improving the production of EFAs by G. candidum in cheese.

Dietary pattern regulates fatty acid desaturase 1 gene expression in Indian pregnant women to spare overall long chain polyunsaturated fatty acids levels.

The aim of this study was to determine if the dietary pattern of pregnant women has any compensatory effect on the fatty acid desaturase (FADS) gene expression, thus enhancing the conversion of precursors to long chain polyunsaturated fatty acids (LCPUFA) to spare the overall LCPUFA levels. The dietary intake of plant-based precursor polyunsaturated fatty acids (PUFA) influences circulating levels of LCPUFA. We hypothesized that low LCPUFA diets during pregnancy would compensate by higher expression of FADS genes to enhance the conversion of precursors to LCPUFA to spare the overall LCPUFA levels. Seventy-five pregnant women were enrolled during the last trimester of pregnancy based on the eligibility and exclusion criteria. Maternal LCPUFA in plasma, expression of FADS1 and FADS2 genes, FADS2 Indel genotype status and neonate birth weight were studied.In the vegetarian group (n = 25), plasma α-linolenic acid (ALA) but not linoleic acid (LA) was significantly lower (p < 0.05) than the non-vegetarian group (n = 50). No significant differences were found for arachidonic acid (AA) or docosahexaenoic acid (DHA) levels. FADS1 expression was significantly higher in the vegetarian group compared to the non-vegetarian group. There was no significant difference in the birth weight of the neonates between two groups. No significant correlation was observed between FADS2 Indel genotype and birth weight. Our small sample size study demonstrated an increase FADS1expression during pregnancy in vegetarian pregnant women that may have contributed to the maintenance of AA, eicosapentaenoic acid and DHA levels thereby ensuring that the overall LCPUFA levels of the neonate is not compromised.

The association of fatty acid desaturase gene polymorphisms on long-chain polyunsaturated fatty acid composition in Indonesian infants

ABSTRACTBackgroundAdequate availability of long-chain polyunsaturated fatty acids (LC-PUFAs) is important for human health from pregnancy to adulthood. Previous studies on fatty acid desaturase (FADS) gene single-nucleotide polymorphisms (SNPs) have been performed predominantly in Western populations and showed that FADS SNPs had a marked impact on LC-PUFA composition in blood and tissues.ObjectivesWe aimed to investigate the influence of fetal FADS genotypes on LC-PUFA composition in umbilical artery plasma lipids in Indonesian infants.DesignWe performed a cross-sectional study to assess for these associations.ResultsA total of 12 cord plasma n–6 (ω-6) and n–3 (ω-3) fatty acids were analyzed for associations with 18 FADS gene cluster SNPs from 390 women with single parturition from the Indonesian Prospective Study of Atopic Dermatitis in Infants (ISADI). Fetal FADS genotypes influenced cord plasma LC-PUFA composition, but, in contrast to previous studies from Western populations, the quantitatively predominant SNPs were associated with lower LC-PUFA content.ConclusionTo our knowledge, this study was the first in South East Asia on FADS genotypes and arterial cord blood fatty acids to show an association between fetal LC-PUFA composition and fetal FADS SNPs. The FADS genotype distribution differs markedly between different geographical populations. This trial was registered at clinicaltrials.gov as NCT02401178.

Functional characterization of a Δ6 fatty acid desaturase gene and its 5′-upstream region cloned from the arachidonic acidrich microalga Myrmecia incisa Reisigl (Chlorophyta)

It is suggested that Δ6 fatty acid desaturase (FAD) plays a critical role in the biosynthesis of polyunsaturated fatty acids in plants and microalgae. But why does it adapt to the changed environments such as nitrogen starvation is seldom understood. One Δ6 FAD gene ( MiD6fad ) from an arachidonic acidrich microalga Myrmecia incisa Reisigl (Chlorophyta) was first heterologously expressed in Saccharomyces cerevisiae for the identification of function. The fatty acid profile of transgenic yeast detected by gas chromatography-mass spectrometry illustrated that the enzyme MiD6FAD could convert linoleic and α - linolenic acids to γ-linolenic and stearidonic acids, respectively, demonstrating that MiD6fad encoded a Δ6 FAD. A 1 965-bp fragment of the cloned 2 347-bp 5′-upstream region of MiD6fad was next subcloned and fused upstream with green fluorescent protein (GFP) gene to replace the GAL1 promoter of the vector pYES2. The generated construct was transformed into S. cerevisiae for function determination. Confocal microscopic images of the transformed line illustrated that this inserted fragment could drive GFP expression, which was further verified by fluorescence intensity quantification and Western blot analysis using anti- GFP antibody. The conversion efficiency (approximately 2% - 3%) of MiD6FAD was much lower than the reported ω 3 FAD and Δ6 elongase in this microalga, suggesting that MiD6FAD catalysed the possible ratelimiting step for ArA biosynthesis. The presence of several putative cis -acting regulatory elements in this identified promoter sheds new light on the regulation mechanism research of Δ6 FAD transcription for the ArA production in M. incisa in changing environmental factors.

Variation of fatty acid desaturation in response to different nitrate levels in Auxenochlorella pyrenoidosa.

Microalgae are promising feedstocks for biodiesel, where the high proportion of monounsaturated fatty acid such as oleic acid (C18:1) is preferred. To regulate fatty acid desaturation in microalgae, the relationship among nitrate concentration, fatty acid composition and the expression levels of desaturase genes was explored. Dynamic variations of fatty acid profiles suggested nitrate could induce desaturation of C18 fatty acids. The content of C18:1 in Auxenochlorella pyrenoidosa was 30.88% at 0 g l−1 nitrate concentration compared with 0.48% at 1.5 g l−1. The expressions of relative delta-9, 12 and 15 fatty acid desaturase genes (Δ9, Δ12 and Δ15FADs) were further investigated. The 330% upregulated expression of Δ9FAD in logarithmic phase at 0 g l−1 resulted in C18:1 accumulation. Moreover, nitrate replenishment caused a sharp reduction of C18:1 from 34.79% to 0.22% and downregulation of Δ9FAD expression to 1% of the nitrate absence level, indicating the pivotal role of Δ9FAD in C18:1 accumulation. Finally, overexpression of Δ9FAD in Escherichia coli and Saccharomyces cerevisiae resulted in an increase of C18:1, confirming its ability of desaturating C18:0. The results could provide a new approach and scientific guidance for the improvement of biodiesel quality and industrialization of high-valued chemicals by means of metabolic engineering.

Genome-wide identification of the fatty acid desaturases gene family in four Aspergillus species and their expression profile in Aspergillus oryzae

Fatty acid desaturases play a key role in producing polyunsaturated fatty acids by converting single bonds to double bonds. In the present study, a total of 13, 12, 8 and 8 candidate fatty acid desaturases genes were identified in the Aspergillus oryzae, Aspergillus flavus, Aspergillus fumigatus and Aspergillus nidulans genomes through database searches, which were classified into five different subfamilies based on phylogenetic analysis. Furthermore, a comprehensive analysis was performed to characterize conserved motifs and gene structures, which could provide an intuitive comprehension to learn the relationship between structure and functions of the fatty acid desaturases genes in different Aspergillus species. In addition, the expression pattern of 13 fatty acid desaturases genes of A. oryzae was tested in different growth stages and under salt stress treatment. The results revealed that the fatty acid desaturases genes in A. oryzae were highly expressed in adaptive phase growth and up-regulated under salt stress treatment. This study provided a better understanding of the evolution and functions of the fatty acid desaturases gene family in the four Aspergillus species, and would be useful for seeking methods to improve the production of unsaturated fatty acids and enhance efforts for the genetic improvement of strains to adapt to the complex surrounding environment.

Membrane physical state and stress regulation in Synechocystis: fluidizing alcohols repress fatty acid desaturation.

Cyanobacteria are prokaryotic photosynthetic organisms widely used in biotechnology, photosynthesis and abiotic stress research. There are several cyanobacterial strains modified to produce biofuels, but the influence of alcohols on cyanobacterial cell physiology is poorly understood. Here, we conducted a systematic study of the effects of nine primary aliphatic alcohols and an aromatic benzyl alcohol on both membrane physical state and the expression of genes for fatty acid desaturases (FADs) in a model cyanobacterium Synechocystis sp. strain PCC 6803. Hexan-1-ol was found to have the most membrane fluidizing action among all alcohols studied, with its efficiency correlating with both duration of treatment and alcohol concentration. A prolonged exposure to alcohol results in a continuous loss of unsaturated fatty acids (FAs) followed by cell death, an undesired challenge that should be considered in cyanobacterial biotechnology. We suggest that membrane fluidization is the key component in alcohol stress causing inactivation of FADs and resulting in a lethal depletion of unsaturated FAs. Due to the most pronounced effects of alcohol- and heat-induced membrane fluidization on desB encoding a terminal ω3-FAD, we propose to call desB a 'viscosity gene' in analogy to heat-induced 'fluidity gene' hspA.

Dietary Polyunsaturated Fatty Acid Supplementations Could Significantly Promote the Δ6 Fatty Acid Desaturase and Fatty Acid Elongase Gene Expression, Long Chain Polyunsaturated Fatty Acids, and Growth of Juvenile Cobia

The experiment investigates the effect of different dietary polyunsaturated fatty acid (PUFA) supplementations on the gene expression of key enzymes for biosynthesizing long chain PUFA (LC-PUFA), LC-PUFA formation, and growth performance of juvenile cobia (Rachycentron canadum). It is found that dietary linolenic (LNA) and linoleic acids (LA) supplementations could significantly promote the expression of △6 fatty acid desaturase (FADS2) and fatty acid elongase (ELOVL5) genes in the brain, liver, and muscle of the juveniles; different dietary PUFA supplementations can significantly promote the growth performance and increase the LC-PUFA in the above organs/tissues of the juveniles; the optimum n-3/n-6PUFA ratio was 0.63 in SO + FO diet, in which n-6PUFAs was 27.4% and n-3PUFAs 17.4% in the diet, with the supplementation of 3% fish oil and 3% safflower oil. Practical Applications: Cobia is a new species with high econimic value and is developing rapidly. The LC-PUFA can promote the development, functions of nerves and cell members in fish juveniles, which are an important period of fish. Can juvenile cobia biosynthesize LC-PUFA from C18:2n-6 and C18:3n-3? The results show that dietary LNA and LA supplementations significantly promoted the FADS2 and ELOVL5 gene expression, fatty acids, and growth of cobia juveniles. Dietary LNA and LA supplementations could significantly promote the expression of Δ6 fatty acid desaturase (FADS2) and fatty acid elongase (ELOVL5) genes in the brain, liver, and muscle of juvenile cobia.