scholarly journals A High-Throughput Fatty Acid Profiling Screen Reveals Novel Variations in Fatty Acid Biosynthesis in Chlamydomonas reinhardtii and Related Algae

2014 ◽  
Vol 13 (11) ◽  
pp. 1431-1438 ◽  
Author(s):  
Erin L. Pflaster ◽  
Michael J. Schwabe ◽  
Joyanne Becker ◽  
Melissa S. Wilkinson ◽  
Ashley Parmer ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Fatty Acid Biosynthesis ◽  
Methyl Esters ◽  
Screening Method ◽  
Laboratory Strain ◽  
Gene Encoding ◽  
Preparation Methods ◽  
Content Type ◽  
Species Specific

ABSTRACTAnalysis of fatty acid methyl esters (FAMEs) by gas chromatography (GC) is a common technique for the quantitative and qualitative analysis of acyl lipids. Methods for FAME preparation are typically time-consuming and labor-intensive and require multiple transfers of reagents and products between reaction tubes and autosampler vials. In order to increase throughput and lower the time and materials costs required for FAME preparation prior to GC analysis, we have developed a method in which 10-to-20-mg samples of microbial biomass are transferred to standard GC autosampler vials, transesterified using an emulsion of methanolic trimethylsulfonium hydroxide and hexane, and analyzed directly by GC without further sample handling. This method gives results that are essentially identical to those obtained by the more labor- and material-intensive FAME preparation methods, such as transmethylation with methanolic HCl. We applied this method to the screening of laboratory and environmental isolates of the green algaChlamydomonasfor variations in fatty acid composition. This screening method facilitated two novel discoveries. First, we identified a common laboratory strain ofC. reinhardtii, CC-620, completely lacking all ω-3 fatty acids normally found in this organism and showed that this strain contains an inactivating mutation in the CrFAD7 gene, encoding the sole ω-3 desaturase activity in this organism. Second, we showed that some species ofChlamydomonasmake Δ6-unsaturated polyunsaturated fatty acids (PUFA) rather than the Δ5 species normally made by the previously characterized laboratory strains ofChlamydomonas, suggesting that there is species-specific variation in the regiospecificity and substrate selectivity of front-end desaturases in this algal genus.

scholarly journals Type II Fatty Acid Biosynthesis Is Essential for Plasmodium falciparum Sporozoite Development in the Midgut of Anopheles Mosquitoes

2013 ◽  
Vol 13 (5) ◽  
pp. 550-559 ◽  
Author(s):  
Ben C. L. van Schaijk ◽  
T. R. Santha Kumar ◽  
Martijn W. Vos ◽  
Adam Richman ◽  
Geert-Jan van Gemert ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Plasmodium Falciparum ◽  
Fatty Acid ◽  
Fatty Acid Biosynthesis ◽  
Blood Stage ◽  
Acid Biosynthesis ◽  
Asexual Blood Stage ◽  
Liver Stage ◽  
Content Type ◽  
Fas Ii

ABSTRACT The prodigious rate at which malaria parasites proliferate during asexual blood-stage replication, midgut sporozoite production, and intrahepatic development creates a substantial requirement for essential nutrients, including fatty acids that likely are necessary for parasite membrane formation. Plasmodium parasites obtain fatty acids either by scavenging from the vertebrate host and mosquito vector or by producing fatty acids de novo via the type two fatty acid biosynthesis pathway (FAS-II). Here, we study the FAS-II pathway in Plasmodium falciparum , the species responsible for the most lethal form of human malaria. Using antibodies, we find that the FAS-II enzyme FabI is expressed in mosquito midgut oocysts and sporozoites as well as liver-stage parasites but not during the blood stages. As expected, FabI colocalizes with the apicoplast-targeted acyl carrier protein, indicating that FabI functions in the apicoplast. We further analyze the FAS-II pathway in Plasmodium falciparum by assessing the functional consequences of deleting fabI and fabB/F . Targeted deletion or disruption of these genes in P. falciparum did not affect asexual blood-stage replication or the generation of midgut oocysts; however, subsequent sporozoite development was abolished. We conclude that the P. falciparum FAS-II pathway is essential for sporozoite development within the midgut oocyst. These findings reveal an important distinction from the rodent Plasmodium parasites P. berghei and P. yoelii , where the FAS-II pathway is known to be required for normal parasite progression through the liver stage but is not required for oocyst development in the Anopheles mosquito midgut.

Fatty acid composition of lipids in immature cattle, pig and sheep oocytes with intact zona pellucida

Reproduction ◽  
2000 ◽  
pp. 163-170 ◽  
Author(s):  
TG McEvoy ◽  
GD Coull ◽  
PJ Broadbent ◽  
JS Hutchinson ◽  
BK Speake
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Zona Pellucida ◽  
Acid Content ◽  
Methyl Esters ◽  
Saturated Fatty Acids ◽  
Fatty Acid Content ◽  
Linoleic Acid Content ◽  
Triglyceride Fraction ◽  
Species Specific

Cattle, pig and sheep oocytes isolated from healthy cumulus-oocyte complexes were pooled, within species, to provide samples of immature denuded oocytes with intact zona pellucida (n = 1000 per sample) for determination of fatty acid mass and composition in total lipid, constituent phospholipid and triglyceride. Acyl-containing lipid extracts, transmethylated in the presence of a reference penta-decaenoic acid (15:0), yielded fatty acid methyl esters which were analysed by gas chromatograph. Mean (+/- SEM) fatty acid content in samples of pig oocytes (161 +/- 18 micrograms per 1000 oocytes) was greater than that in cattle (63 +/- 6 micrograms; P < 0.01) and sheep oocytes (89 +/- 7 micrograms; P < 0.05). Of 24 fatty acids detected, palmitic (16:0; 25-35%, w/w), stearic (18:0; 14-16%) and oleic (18:1n-9; 22-26%) acids were most prominent in all three species. Saturated fatty acids (mean = 45-55%, w/w) were more abundant than mono- (27-34%) or polyunsaturates (11-21%). Fatty acids of the n-6 series, notably linoleic (18:2n-6; 5-8%, w/w) and arachidonic acid (20:4n-6; 1-3%), were the most abundant polyunsaturates. Phospholipid consistently accounted for a quarter of all fatty acids in the three species, but ruminant oocytes had a lower complement of polyunsaturates (14-19%, w/w) in this fraction than pig oocytes (34%, w/w) which, for example, had a three- to fourfold greater linoleic acid content. An estimated 74 ng of fatty acid was sequestered in the triglyceride fraction of individual pig oocytes compared with 23-25 ng in ruminant oocytes (P < 0.01). It is concluded that the greater fatty acid content of pig oocytes is primarily due to more abundant triglyceride reserves. Furthermore, this species-specific difference, and that in respect of polyunsaturated fatty acid reserves, may underlie the contrasting chilling, culture and cryopreservation sensitivities of embryos derived from pig and ruminant (cattle, sheep) oocytes.

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 The same rat Δ6-desaturase not only acts on 18- but also on 24-carbon fatty acids in very-long-chain polyunsaturated fatty acid biosynthesis

2002 ◽  
Vol 364 (1) ◽  
pp. 49-55 ◽  
Author(s):  
Sabine D'ANDREA ◽  
Hervé GUILLOU ◽  
Sophie JAN ◽  
Daniel CATHELINE ◽  
Jean-Noël THIBAULT ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Polyunsaturated Fatty Acid ◽  
Unsaturated Fatty Acids ◽  
Fatty Acid Biosynthesis ◽  
Chain Polyunsaturated Fatty Acid ◽  
Acid Biosynthesis ◽  
Highly Unsaturated Fatty Acids ◽  
Long Chain ◽  
Δ6 Desaturase

The recently cloned Δ6-desaturase is known to catalyse the first step in very-long-chain polyunsaturated fatty acid biosynthesis, i.e. the desaturation of linoleic and α-linolenic acids. The hypothesis that this enzyme could also catalyse the terminal desaturation step, i.e. the desaturation of 24-carbon highly unsaturated fatty acids, has never been elucidated. To test this hypothesis, the activity of rat Δ6-desaturase expressed in COS-7 cells was investigated. Recombinant Δ6-desaturase expression was analysed by Western blot, revealing a single band at 45kDa. The putative involvement of this enzyme in the Δ6-desaturation of C24:5n-3 to C24:6n-3 was measured by incubating transfected cells with C22:5n-3. Whereas both transfected and non-transfected COS-7 cells were able to synthesize C24:5n-3 by elongation of C22:5n-3, only cells expressing Δ6-desaturase were also able to produce C24:6n-3. In addition, Δ6-desaturation of [1-14C]C24:5n-3 was assayed invitro in homogenates from COS-7 cells expressing Δ6-desaturase or not, showing that Δ6-desaturase catalyses the conversion of C24:5n-3 to C24:6n-3. Evidence is therefore presented that the same rat Δ6-desaturase catalyses not only the conversion of C18:3n-3 to C18:4n-3, but also the conversion of C24:5n-3 to C24:6n-3. A similar mechanism in the n-6 series is strongly suggested.

scholarly journals Fatty Acids Profiles of Stipe and Blade from the Norwegian Brown Macroalga Laminaria hyperborea, Using Off-Line SPE and GC-MS

2016 ◽  
Author(s):  
Lena Oksdøl Foseid ◽  
Hanne Devle ◽  
Yngve Stenstrøm ◽  
Carl Fredrik Naess-Andresen ◽  
Dag Ekeberg
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Polyunsaturated Fatty Acids ◽  
Neutral Lipids ◽  
Methyl Esters ◽  
Lipid Fraction ◽  
Environmental Benefits ◽  
Laminaria Hyperborea ◽  
Alpha Linolenic Acid ◽  
Lipid Fractions

A thorough analysis and comparison of the fatty acid profiles of stipe and blade from Laminaria hyperborea, a kelp species found in the northern Atlantic, is presented. Lipids were extracted and fractionated into neutral lipids, free fatty acids and polar lipids, then derivatized to fatty acid methyl esters prior to GC-MS analysis. A total of 42 fatty acids were identified and quantified, including the n-3 fatty acids &alpha;-linolenic acid, stearidonic acid and eicosapentaenoic acid. An n-6/n-3 ratio of 0.8:1 was found in blade and 3.5:1 in stipe, respectively. The ratios vary between the lipid fractions within stipe and blade, with the lowest ratio in the polar lipid fraction of blade. The fatty acid amounts are higher in blade than in stipe, and the highest amounts of n-3 fatty acids are found within the neutral lipid fractions. The amounts of polyunsaturated fatty acids are 3.4 times higher in blade than stipe. This study highlights the compositional differences between the lipid fractions of stipe and blade from L. hyperborea. The amount of polyunsaturated fatty acids, compared to saturated- and monounsaturated fatty acids, as well as the n-6/n-3-ratio, is known to influence human health. In the pharmaceutical, food, and feed industries this can be of importance for production and sale of different health products. Additionally, lipids are today among the unused by products of alginate production, exploiting this material for commercial interest should give both economical and environmental benefits.

scholarly journals Inclusion of rapeseed and pumpkin seed cakes in diets for Murciano-Granadina goats alters the fatty acid profile of milk

2021 ◽  
Vol 51 (2) ◽  
pp. 262-270
Author(s):  
I.M. Boldea ◽  
C. Dragomir ◽  
M.A. Gras ◽  
M. Ropotă
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Fatty Acid Profile ◽  
Unsaturated Fatty Acids ◽  
Methyl Esters ◽  
Control Diet ◽  
Dietary Lipids ◽  
Italian Ryegrass ◽  
Pumpkin Seed ◽  
Lactating Goats

The objective of this research was to assess the effects of including oil-rich feedstuffs in diets for lactating goats on the fatty acid (FA) profile of their milk. Thirty-six Murciano-Granadina goats were randomly assigned to three treatment groups, namely a control diet (CTRL), a diet based on whole rapeseed (RS), and a diet based on pumpkin seed cake (PSC). The diets were composed of 1 kg hay (70 % Italian ryegrass, 30% alfalfa) and 1.24 kg concentrate, and were formulated to be isoenergetic and isonitrogenous. Milk yield and its contents of protein, fat and lactose did not differ significantly among the groups. However, including oil-rich feeds in the diet altered the fatty acid profile of the milk significantly, decreasing its saturated fatty acid (SFA) content and increasing its content of unsaturated fatty acids (UFAs). Effects on polyunsaturated fatty acids (PUFAs), conjugated linoleic acid (CLA), and the n-6 to n-3 ratio depended on the source of dietary lipids. The PSC augmented diet increased the relative amount of PUFAs and fatty acid methyl esters (FAME) in milk (+25 %) significantly In comparison with CTRL, whereas the RS diet produced a limited and statistically insignificant increase (+7.5%). The concentration of CLA was higher in milk from does fed the PSC diet, whereas the n-6 to n-3 ratio was lower in milk from does fed RS. These preliminary results form the basis for developing premium dairy products that are enriched in fatty acids that are more favourable for human health.

Lipids of the fin whale (Balaenoptera physalus) from North Atlantic waters. IV. Fin whale milk

1968 ◽  
Vol 46 (3) ◽  
pp. 197-203 ◽  
Author(s):  
R. G. Ackman ◽  
C. A. Eaton ◽  
S. N. Hooper
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Polyunsaturated Fatty Acids ◽  
Total Lipid ◽  
Iodine Value ◽  
North Atlantic ◽  
Methyl Esters ◽  
Short Chain Fatty Acids ◽  
Fin Whale ◽  
Hydrolysis Of

Fatty acid compositions were determined for total lipid (17.5% of the milk and > 95% triglycerides), 2-monoglyceride obtained by enzymatic hydrolysis of isolated triglyceride, and isolated phospholipid (~1% of total lipids). The total lipid fatty acids of the milk had a composition similar to fin whale depot fat but were enriched in hexadecanoic acid and polyunsaturated fatty acids at the expense of monoethylenic acids; correspondingly the iodine value of 136 (methyl esters) was higher than the normal range (105–120) of North Atlantic fin whale blubber oils. Over 80% of the fatty acids in the 2-position of the triglycerides were accounted for by relatively short chain fatty acids, especially hexadecanoic (54.6%), tetradecanoic (13.7%), and hexadecenoic (11.2%), so that the ester iodine value was only 48. The milk phospholipids had a fatty acid composition basically similar to that of liver phospholipids (methyl ester iodine value 120) with somewhat more polyunsaturated fatty acids and accordingly an iodine value of 144 for methyl esters.

scholarly journals Proteomic Analysis of the Function of a Novel Cold-Regulated Multispanning Transmembrane Protein COR413-PM1 in Arabidopsis

2018 ◽  
Vol 19 (9) ◽  
pp. 2572 ◽  
Author(s):  
Chen Su ◽  
Kai Chen ◽  
Qingqian Ding ◽  
Yongying Mou ◽  
Rui Yang ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Plasma Membrane ◽  
Fatty Acid ◽  
Proteomic Analysis ◽  
Fatty Acid Biosynthesis ◽  
Transmembrane Protein ◽  
Sugar Metabolism ◽  
Freezing Stress ◽  
Plant Tolerance ◽  
Phosphoribosyl Pyrophosphate

The plasma membrane is the first subcellular organ that senses low temperature, and it includes some spanning transmembrane proteins that play important roles in cold regulation. COR413-PM1 is a novel multispanning transmembrane cold-regulated protein; however, the related functions are not clear in Arabidopsis. We found the tolerance to freezing stress of cor413-pm1 was lower than wild-type (WT). A proteomics method was used to analyze the differentially abundant proteins (DAPs) between cor413-pm1 and WT. A total of 4143 protein groups were identified and 3139 were accurately quantitated. The DAPs associated with COR413-PM1 and freezing treatment were mainly involved in the metabolism of fatty acids, sugars, and purine. Quantitative real-time PCR (qRT-PCR) confirmed the proteomic analysis results of four proteins: fatty acid biosynthesis 1 (FAB1) is involved in fatty acid metabolism and might affect the plasma membrane structure; fructokinase 3 (FRK3) and sucrose phosphate synthase A1 (SPSA1) play roles in sugar metabolism and may influence the ability of osmotic adjustment under freezing stress; and GLN phosphoribosyl pyrophosphate amidotransferase 2 (ASE2) affects freezing tolerance through purine metabolism pathways. In short, our results demonstrate that the multispanning transmembrane protein COR413-PM1 regulates plant tolerance to freezing stress by affecting the metabolism of fatty acids, sugars, and purine in Arabidopsis.

212 Fatty Acid Analysis and Composition in Meat by Mass Spectrometry

2021 ◽  
Vol 99 (Supplement_3) ◽  
pp. 111-112
Author(s):  
Thu Dinh
Keyword(s):  
Mass Spectrometry ◽  
Fatty Acids ◽  
Fatty Acid ◽  
Methyl Esters ◽  
Meat Products ◽  
Chemical Properties ◽  
Fatty Acid Analysis ◽  
Animal Fats ◽  
Wide Range ◽  
Mass Spectrometry Technology

Abstract Fatty acids determine the physical and chemical properties of fats. Animal fats, regardless of species, have more saturated and monounsaturated than polyunsaturated fatty acids. The major fatty acids in meat are palmitic (16:0), stearic (18:0), palmitoleic (16:1), oleic (18:1), linoleic (18:2), and linolenic (18:3) acids, among which oleic acid is the most predominant. Arachidonic acid (20:4 cis 5,8,11,14) is an essential fatty acid only found in animal fats and can be used as a quality control indicator in the fatty acid analysis. Fatty acid analysis has been traditionally performed by gas chromatography (GC) of volatile fatty acid derivatives, prominently the methyl esters, and flame ionization detection (FID), in which the carbon chain of fatty acids is degraded to the formylium ion CHO+. The FID is very sensitive and is the most widely used detection method for GC, providing a linear response, i.e., peak area, over a wide range of concentrations. Researchers have been used the FID peak area to calculate the percentages of fatty acids. However, the FID is a “carbon counter” and relies on the “equal per carbon” rule; therefore, at the same molar concentration, fatty acids with a different number of carbons produce different peak areas. The recent development of mass spectrometry technology has improved the specificity of fatty acid detection. Specific target and qualifier ions provide better identification and more accurate quantification of fatty acid concentrations. Although fatty acids can be identified through comparing ion fragmentation with various databases, authentic standards are needed for quantification purposes. Using mass spectrometry, more than 50 fatty acids have been identified in meat samples. Some branched-chain fatty acids may have flavor, safety, and shelf life implications in meat products.

Analysis of Lipids Produced by Microalgae Isolated from the Area around Okinawa, Japan

2017 ◽  
pp. 222-233
Author(s):  
Shinya Ikematsu ◽  
Ipputa Tada ◽  
Yasuma Nagasaki
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Palmitic Acid ◽  
Methyl Esters ◽  
Dry Weight ◽  
Botryococcus Braunii ◽  
Biodiesel Fuel ◽  
High Lipid ◽  
Institute Of Technology ◽  
Petroleum Reserves

Petroleum reserves have been decreasing in recent years and microalgae are attractive as a potential source of new biomass petroleum. Microalgae are unicellar microscopic algae and most species microalgae produce lipids. In particular, Botryococcus braunii produces large amount of lipids found with nearly 70% on the basis of the dry weight. This chapter reviews high lipid-producing microalgae found from Okinawa area around National Institute of Technology, Okinawa College (NIT, Okinawa). The microalgae collected were isolated on an AF-6 agar plates, and incubated in AF-6 medium. The fatty acids were extracted from the algae, converted into fatty acid methyl esters, and analysed by GC/MS. As a result, two microalgae strains were identified that the produced fatty acids was loaded in the algae with nearly 20% in the dry weight base. In addition, these two microalgae strains produced palmitic acid as nearly 40% of the total produced lipids. Therefore, the two microalga strains isolated are potentially and highly efficient for the organisms applied for the production of biodiesel fuel.

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