scholarly journals Cytochrome b 5 Coexpression Increases Tetrahymena thermophila Δ6 Fatty Acid Desaturase Activity in Saccharomyces cerevisiae

2013 ◽  
Vol 12 (6) ◽  
pp. 923-931 ◽  
Author(s):  
Jeremy L. Dahmen ◽  
Rebecca Olsen ◽  
Deirdre Fahy ◽  
James G. Wallis ◽  
John Browse
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Fatty Acids ◽  
Fatty Acid ◽  
Protein Interactions ◽  
Desaturase Activity ◽  
Tetrahymena Thermophila ◽  
Fatty Acid Desaturase ◽  
Vital Role ◽  
Content Type ◽  
Split Ubiquitin

ABSTRACT Very-long-chain polyunsaturated fatty acids such as arachidonic, eicosapentaenoic, and docosahexaenoic acids, are important to the physiology of many microorganisms and metazoans and are vital to human development and health. The production of these and related fatty acids depends on Δ6 desaturases, the final components of an electron transfer chain that introduces double bonds into 18-carbon fatty acid chains. When a Δ6 desaturase identified from the ciliated protist Tetrahymena thermophila was expressed in Saccharomyces cerevisiae cultures supplemented with the 18:2 Δ9,12 substrate, only 4% of the incorporated substrate was desaturated. Cytochrome b 5 protein sequences identified from the genome of T. thermophila included one sequence with two conserved cytochrome b 5 domains. Desaturation by the Δ6 enzyme increased as much as 10-fold when T. thermophila cytochrome b 5 s were coexpressed with the desaturase. Coexpression of a cytochrome b 5 from Arabidopsis thaliana with the Δ6 enzyme also increased desaturation. A split ubiquitin growth assay indicated that the strength of interaction between cytochrome b 5 proteins and the desaturase plays a vital role in fatty acid desaturase activity, illustrating the importance of protein-protein interactions in this enzyme activity.

scholarly journals Heterologous Production of Dihomo-γ-Linolenic Acid in Saccharomyces cerevisiae

2007 ◽  
Vol 73 (21) ◽  
pp. 6965-6971 ◽  
Author(s):  
Hisashi Yazawa ◽  
Hitoshi Iwahashi ◽  
Yasushi Kamisaka ◽  
Kazuyoshi Kimura ◽  
Tsunehiro Aki ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Fatty Acids ◽  
Fatty Acid ◽  
Linolenic Acid ◽  
Acid Content ◽  
Kluyveromyces Lactis ◽  
Fatty Acid Content ◽  
Fatty Acid Desaturase ◽  
Dry Weight ◽  
Medium Composition

ABSTRACT To make dihomo-γ-linolenic acid (DGLA) (20:3n-6) in Saccharomyces cerevisiae, we introduced Kluyveromyces lactis Δ12 fatty acid desaturase, rat Δ6 fatty acid desaturase, and rat elongase genes. Because Fad2p is able to convert the endogenous oleic acid to linoleic acid, this allowed DGLA biosynthesis without the need to supply exogenous fatty acids on the media. Medium composition, cultivation temperature, and incubation time were examined to improve the yield of DGLA. Fatty acid content was increased by changing the medium from a standard synthetic dropout medium to a nitrogen-limited minimal medium (NSD). Production of DGLA was higher in the cells grown at 15�C than in those grown at 20�C, and no DGLA production was observed in the cells grown at 30�C. In NSD at 15�C, fatty acid content increased up until day 7 and decreased after day 10. When the cells were grown in NSD for 7 days at 15�C, the yield of DGLA reached 2.19 μg/mg of cells (dry weight) and the composition of DGLA to total fatty acids was 2.74%. To our knowledge, this is the first report describing the production of polyunsaturated fatty acids in S. cerevisiae without supplying the exogenous fatty acids.

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

Nutrients ◽  
2019 ◽  
Vol 11 (3) ◽  
pp. 602 ◽  
Author(s):  
Isabel Salas Lorenzo ◽  
Aida Chisaguano Tonato ◽  
Andrea de la Garza Puentes ◽  
Ana Nieto ◽  
Florian Herrmann ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Breast Milk ◽  
Infant Formula ◽  
Early Life ◽  
Desaturase Activity ◽  
Reference Group ◽  
Fatty Acid Desaturase ◽  
Breastfed Infants ◽  
Major Allele

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.

scholarly journals Molecular cloning of Δ9 fatty acid desaturase from the protozoan Tetrahymena thermophila and its mRNA expression during thermal membrane adaptation

1996 ◽  
Vol 317 (1) ◽  
pp. 29-34 ◽  
Author(s):  
Shigeru NAKASHIMA ◽  
Yutong ZHAO ◽  
Yoshinori NOZAWA
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Cold Adaptation ◽  
Desaturase Activity ◽  
Unsaturated Fatty Acids ◽  
Self Regulation ◽  
Control Level ◽  
Fatty Acid Desaturase ◽  
Amblyomma Americanum ◽  
Transcriptional Level

In response to a decrease in its growth temperature, the protozoan Tetrahymena is known to increase the level of unsaturated fatty acids in its membrane phospholipids so as to maintain the correct physical state (fluidity) of the membranes. In this organism, synthesis of unsaturated fatty acids is initiated by Δ9 acyl-CoA desaturase. Our previous studies have shown that, during cold adaptation, the activity of microsomal palmitoyl- and stearoyl-CoA desaturase increases, reaching a maximal level at 2 h after a temperature down-shift to 15 °C. Two hypotheses have been proposed to explain this increase in desaturase activity: (1) self-regulation via a direct effect of reduced membrane fluidity, and (2) induction of desaturase mRNA. However, the precise mechanism is not clearly understood. In order to obtain further insight into the mechanism of regulation of the desaturase, we have isolated a gene that encodes Δ9 fatty acid desaturase from T. thermophila and examined its expression during cold adaptation. The nucleotide sequence indicates that the 1.4 kbp gene encodes a polypeptide of 292 amino acid residues which shows marked sequence similarity to Δ9 acyl-CoA desaturases from other sources, e.g. rat, mouse, Amblyomma americanum and Saccharomyces cerevisiae. This protein has three histidine-cluster motifs (one HXXXXH and two HXXHH), and two hydrophobic regions which are conserved among Δ9 acyl-CoA desaturases. The level of desaturase mRNA was sensitive to decreasing the temperature of the culture media, and was close to maximal immediately after the temperature was shifted down from 35 °C to 15 °C (0.8 °C/min). Thereafter, the amount of mRNA gradually decreased with time, but remained above the control level for at least 5 h. Furthermore, during the course of the cooling process to 15 °C, the increased expression of desaturase mRNA became evident at 27 °C. Nuclear run-on analysis and actinomycin D chase experiments revealed that the elevation of the mRNA level was due to increases in both transcription and mRNA stability. These results suggest that the enhanced desaturase activity is controlled, at least in part, at the transcriptional level.

ω-3 Long-Chain Polyunsaturated Fatty Acids and Fatty Acid Desaturase Activity Ratios as Eventual Endophenotypes for ADHD

2012 ◽  
Vol 19 (11) ◽  
pp. 977-986 ◽  
Author(s):  
Marcela Henríquez-Henríquez ◽  
Sandra Solari ◽  
Gisela Várgas ◽  
Luis Vásquez ◽  
Fidel Allende ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Polyunsaturated Fatty Acids ◽  
Desaturase Activity ◽  
Fatty Acid Desaturase ◽  
Activity Ratios ◽  
Long Chain

scholarly journals Heat stress elicits remodeling in the anther lipidome of peanut

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Zolian S. Zoong Lwe ◽  
Ruth Welti ◽  
Daniel Anco ◽  
Salman Naveed ◽  
Sachin Rustgi ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Heat Stress ◽  
Fatty Acid ◽  
Membrane Lipids ◽  
Fatty Acid Desaturase ◽  
Susceptible Genotype ◽  
Unsaturated Lipid ◽  
Lipid Species ◽  
Lipid Unsaturation ◽  
Fatty Acid Amount

AbstractUnderstanding the changes in peanut (Arachis hypogaea L.) anther lipidome under heat stress (HT) will aid in understanding the mechanisms of heat tolerance. We profiled the anther lipidome of seven genotypes exposed to ambient temperature (AT) or HT during flowering. Under AT and HT, the lipidome was dominated by phosphatidylcholine (PC), phosphatidylethanolamine (PE), and triacylglycerol (TAG) species (> 50% of total lipids). Of 89 lipid analytes specified by total acyl carbons:total carbon–carbon double bonds, 36:6, 36:5, and 34:3 PC and 34:3 PE (all contain 18:3 fatty acid and decreased under HT) were the most important lipids that differentiated HT from AT. Heat stress caused decreases in unsaturation indices of membrane lipids, primarily due to decreases in highly-unsaturated lipid species that contained 18:3 fatty acids. In parallel, the expression of Fatty Acid Desaturase 3-2 (FAD3-2; converts 18:2 fatty acids to 18:3) decreased under HT for the heat-tolerant genotype SPT 06-07 but not for the susceptible genotype Bailey. Our results suggested that decreasing lipid unsaturation levels by lowering 18:3 fatty-acid amount through reducing FAD3 expression is likely an acclimation mechanism to heat stress in peanut. Thus, genotypes that are more efficient in doing so will be relatively more tolerant to HT.

Saccharomyces kluyveri FAD3 encodes an ω3 fatty acid desaturase

Microbiology ◽  
2004 ◽  
Vol 150 (6) ◽  
pp. 1983-1990 ◽  
Author(s):  
Takahiro Oura ◽  
Susumu Kajiwara
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Linoleic Acid ◽  
Linolenic Acid ◽  
Functional Characterization ◽  
Fatty Acid Desaturase ◽  
Fatty Acid Desaturases ◽  
Desaturase Gene ◽  
Saccharomyces Kluyveri ◽  
Fatty Acid Desaturase Gene

Fungi, like plants, are capable of producing the 18-carbon polyunsaturated fatty acids linoleic acid and α-linolenic acid. These fatty acids are synthesized by catalytic reactions of Δ12 and ω3 fatty acid desaturases. This paper describes the first cloning and functional characterization of a yeast ω3 fatty acid desaturase gene. The deduced protein encoded by the Saccharomyces kluyveri FAD3 gene (Sk-FAD3) consists of 419 amino acids, and shows 30–60 % identity with Δ12 fatty acid desaturases of several eukaryotic organisms and 29–31 % identity with ω3 fatty acid desaturases of animals and plants. During Sk-FAD3 expression in Saccharomyces cerevisiae, α-linolenic acid accumulated only when linoleic acid was added to the culture medium. The disruption of Sk-FAD3 led to the disappearance of α-linolenic acid in S. kluyveri. These findings suggest that Sk-FAD3 is the only ω3 fatty acid desaturase gene in this yeast. Furthermore, transcriptional expression of Sk-FAD3 appears to be regulated by low-temperature stress in a manner different from the other fatty acid desaturase genes in S. kluyveri.

Long-chain n-3 polyunsaturated fatty acids contained in liver and muscle tissues of Okinawan long-spine porcupinefish (Diodon holocanthus Linnaeus 1758)

2021 ◽  
Vol ahead-of-print (ahead-of-print) ◽  
Author(s):  
Yutaka Tashiro
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Polyunsaturated Fatty Acids ◽  
Ryukyu Islands ◽  
Okinawa Island ◽  
Content Type ◽  
Muscle Tissues ◽  
The Ryukyu Islands ◽  
Total Fatty Acids ◽  
Long Chain

Purpose This study aimed to analyze the lipid content and fatty acid composition in the liver and muscle of a porcupinefish species inhabiting waters around the Ryukyu Islands to investigate their potential as a source of long-chain n-3 polyunsaturated fatty acids (LC-PUFAs). Design/methodology/approach Porcupinefish were collected along the Okinawa Island coast. The composition of fatty acids and cholesterol in both liver and muscle were analyzed using a gas chromatograph mass spectrometer. Findings The liver of Okinawan long-spine porcupinefish was rich in lipids whose content correlated to the proportion of liver/body weight. Fatty acid compositions in their liver and muscles were similar to each other. LC-PUFAs occupied 44% of total fatty acids, with docosahexaenoic acid (DHA) being the dominant (42%), whereas eicosapentaenoic acid occupied 2.4%. The liver contained 1,690 mg of cholesterol and 14.8 g of DHA per 100 g, whose proportion decreased in summer compared to other seasons (p = 0.036). Originality/value The liver of Okinawan long-spine porcupinefish, which has not yet been commercially used although its non-toxicity is claimed, can be an excellent source of LC-PUFAs, especially DHA, accentuating its potential in food supplements’ production.

scholarly journals Deficiency of Linolenic Acid in Lefad7 Mutant Tomato Changes the Volatile Profile and Sensory Perception of Disrupted Leaf and Fruit Tissue

2006 ◽  
Vol 131 (2) ◽  
pp. 284-289 ◽  
Author(s):  
Mauricio A. Cañoles ◽  
Randolph M. Beaudry ◽  
Chuanyou Li ◽  
Gregg Howe
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Linolenic Acid ◽  
Fatty Acid Desaturase ◽  
Primary Source ◽  
Mutant Line ◽  
Volatile Profile ◽  
Omega 3 ◽  
Line Production ◽  
Fruit Tissue

Six-carbon aldehydes and alcohols formed by tomato (Lycopersicon esculentum Mill.) leaf and fruit tissue following disruption are believed to be derived from the degradation of lipids and free fatty acids. Collectively, these C-6 volatiles comprise some of the most important aroma impact compounds. If fatty acids are the primary source of tomato volatiles, then an alteration in the fatty acid composition such as that caused by a mutation in the chloroplastic omega-3-fatty acid desaturase (ω-3 FAD), referred to as LeFAD7, found in the mutant line of `Castlemart' termed Lefad7, would be reflected in the volatile profile of disrupted leaf and fruit tissue. Leaves and fruit of the Lefad7 mutant had ≈10% to 15% of the linolenic acid (18:3) levels and about 1.5- to 3-fold higher linoleic acid (18:2) levels found in the parent line. Production of unsaturated C-6 aldehydes Z-3-hexenal, Z-3-hexenol, and E-2-hexenal and the alcohol Z-3-hexenol derived from 18:3 was markedly reduced in disrupted leaf and fruit tissue of the Lefad7 mutant line. Conversely, the production of the saturated C-6 aldehyde hexanal and its alcohol, hexanol, were markedly higher in the mutant line. The shift in the volatile profile brought about by the loss of chloroplastic FAD activity in the Lefad7 line was detected by sensory panels at high significance levels (P < 0.0005) and detrimentally affected fruit sensory quality. The ratios and amounts of C-6 saturated and unsaturated aldehydes and alcohols produced by tomato were dependent on substrate levels, suggesting that practices that alter the content of linoleic and linolenic acids or change their ratios can influence tomato flavor.

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