Noemí Ruiz-López

University of Malaga | UMA · Department of Biochemistry and Molecular Biology

Omega-3 (also called n-3) long-chain polyunsaturated fatty acids (≥C20; LC-PUFAs) are of considerable interest, based on clear evidence of dietary health benefits and the concurrent decline of global sources (fish oils). Generating alternative transgenic plant sources of omega-3 LC-PUFAs, i.e. eicosapentaenoic acid (20:5 n-3, EPA) and docosahexaeno...

The synthesis and accumulation of omega-3 long-chain polyunsaturated fatty acids in transgenic Camelina sativa is demonstrated using the so-called alternative pathway. This aerobic pathway is found in a small number of taxonomically unrelated unicellular organisms and utilizes a C18 Δ9-elongase to generate C20 PUFAs. Here, we evaluated four differe...

High biomass crops have recently attracted significant attention as an alternative platform for the renewable production of high energy storage lipids such as triacylglycerol (TAG). While TAG typically accumulates in seeds as storage compounds fuelling subsequent germination, levels in vegetative tissues are generally low. Here, we report the accum...

The global consumption of fish oils currently exceeds one million tonnes, with the natural de novo source of these important fatty acids forming the base of marine foodwebs. Here we describe the first field-based evaluation of a terrestrial source of these essential nutrients, synthesised in the seeds of transgenic Camelina sativa plants via the he...

Las proteínas con un dominio SMP se encuentran evolutivamente conservadas y participan en la formación de puntos de contacto de membrana (MCS, membrane contact sites), unas estructuras donde las membranas de dos orgánulos se encuentran muy cerca pero sin llegar a contactar y que están implicadas en el transporte lipídico entre compartimentos celula...

Plants have developed mechanisms to protect themselves from pathogens and resist their attacks. Among these, clustering of chloroplasts around the nuclear envelope is a phenomenon that has been proposed as a general response to the perception of pathogens, such as viruses and bacteria [1]. Additionally, it has been hypothesized that chloroplast clu...

Membrane contact sites (MCS) are regions where two membranes of different organelles are close but not fused; they coordinate non-vesicular communication between organelles and are involved in a wide variety of physiological functions, including membrane lipid homeostasis. Amongst proteins localized at MCS are those containing a lipid transport dom...

Controlled primary cell wall remodeling allows plant growth under stressful conditions, but how these changes are conveyed to adjust cellulose synthesis is not understood. Here, we identify the TETRATRICOPEPTIDE THIOREDOXIN-LIKE (TTL) proteins as new members of the cellulose synthase complex (CSC) and describe their unique and hitherto unknown dyna...

Controlled primary cell wall remodeling adapts plant growth under stressful conditions, but how these changes are conveyed to adjust cellulose synthesis is unknown. Here, we identify the Tetratricopeptide Thioredoxin-Like (TTL) proteins as new members of the cellulose synthase complex (CSC) and describe their unique and hitherto unknown dynamic ass...

Castor beans accumulate large amounts of triacylglycerols (TAGs) in the seed endosperm. This oil contains hydroxylated ricinoleic levels close to 90%, which is unique among oil seeds. The capacity to accumulate such high levels of such an unusual fatty acids is due to its specific accumulation and channeling. Here, the ability of the castor biosynt...

Non-vesicular lipid transfer at ER and plasma membrane (PM) contact sites (CS) is crucial for the maintenance of membrane lipid homeostasis. Extended synaptotagmins (E-Syts) play a central role in this process as they act as molecular tethers of ER and PM and as lipid transfer proteins between these organelles. E-Syts are proteins constitutively an...

Endoplasmic reticulum-plasma membrane contact sites (ER-PM CS) play fundamental roles in all eukaryotic cells. Arabidopsis thaliana mutants lacking the ER-PM protein tether synaptotagmin1 (SYT1) exhibit decreased plasma membrane (PM) integrity under multiple abiotic stresses such as freezing, high salt, osmotic stress and mechanical damage. Here, w...

Cell function requires the maintenance of membrane lipid homeostasis as changes in cellular environment unbalance this equilibrium. The non-vesicular lipid transfer at endoplasmic reticulum (ER) and plasma membrane (PM) contact sites (CS) is central to restore it. Extended synaptotagmins (E-Syts) are ER proteins that play a central role in this pro...

The enzymes involved in L-ascorbate biosynthesis in photosynthetic organisms (the Smirnoff-Wheeler (SW) pathway) are well established. Here we analyzed their subcellular localizations and potential physical interactions and assessed their role in the control of ascorbate synthesis. Transient expression of C terminal-tagged fusions of SW genes in Ni...

Endoplasmic Reticulum-Plasma Membrane contact sites (ER-PM CS) play fundamental roles in all eukaryotic cells. Arabidopsis mutants lacking the ER-PM protein tether synaptotagmin1 (SYT1) exhibit decreased plasma membrane (PM) integrity under multiple abiotic stresses such as freezing, high salt, osmotic stress and mechanical damage. Here, we show th...

The castor oil plant represents a promising platform to produce oils with industrial applications. However, its use in biotechnology is limited by the absence of a well-established procedure to transform it, and a poor understanding of gene regulation and promoter use in this species. As such, a method has been developed to express proteins or hair...

Wax esters (WEs) and steryl esters (SEs) are minor components of sunflower oils formed by the esterification of long chain fatty alcohols and sterols to fatty acids. These compounds have similar carbon numbers and polarities making them difficult to separate using conventional chromatographic methods. In this study, electrospray ionisation-tandem m...

Recent studies suggest that the use of vegetable oils at expense of fish oil in aquaculture feeds might have potential negative effects on fish redox homeostasis and adiposity. Resveratrol (RESV) is a lipid-soluble phytoalexin present in fruits and vegetables with proven in vivo antioxidant function in animals. The present study aims to assess the...

Table S1 Acyl‐CoA profiling of developing Camelina Seeds Table S2 Molecular distillation results Table S3 Oil winterisation results Table S4a Physico‐chemical properties of saturated wax esters Table S4b Physico‐chemical properties of unsaturated wax esters

Figure S1 (a) Wax ester content in the seeds of wild‐type and transgenic Camelina sativa lines expressing different combinations of wax biosynthetic and fatty acid modifying enzymes. (b) Wax ester profile in the seeds of T3 transgenic lines expressing either MaMa14 (left panel) or MoMa14 (right panel) enzyme combinations. Figure S2 Wax ester compo...

The functional characterisation of wax biosynthetic enzymes in transgenic plants has opened the possibility of producing tailored wax esters (WEs) in the seeds of a suitable host crop. In this study, in addition to systematically evaluating a panel of WE biosynthetic activities, we have also modulated the acyl-CoA substrate pool, through the co-exp...

Background: Fish currently supplies only 40% of the eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) required to allow all individuals globally to meet the minimum intake recommendation of 500 mg/d. Therefore, alternative sustainable sources are needed. Objective: The main objective was to investigate the ability of genetically engineered...

Figure S1 Differentiation of ETA from ARA and juniperonic acid. Figure S2 Range of ETA and EPA content in single seeds from AP5#1_5. Figure S3 Comparison of fatty profiles for fish oil, Camelina and AP5#1_5.

Table S1 Single seed analysis – fatty acid composition of individual seeds of line AP5#1_5.

The nutritional and functional characteristics of dietary fat are related to the fatty acid (FA) composition and its positional distribution in the triacylglycerol (TAG) fraction. Atlantic salmon is an important source of healthy long chain omega 3 FA (particularly, eicosapentaenoic (EPA) and docoxahexaenoic (DHA) acids). However, the impact of lip...

Supplementary data Supplementary Table 1 Total seed fatty acid composition of selected event DHA-5C♯33_13 over multiple generations.

Supplementary data Supplementary Table 3 Recorded weather conditions on the Rothamsted Experimental Farm during the period of the GM field trial. The sowing and harvest dates are also provided, as are the conditions in the glasshouse.

Supplementary data Supplementary Figure 1 Description of the location and layout of the GM C. sativa trial. Supplementary Figure 2 EPA and DHA levels present in GM seeds from plants grown in either field plots or glasshouse. Statistical analysis is as described in Methods. Supplementary Figure 3 Seed moisture content for field-grown and glasshouse...

Supplementary data Supplementary Table 2 EPA and DHA content (total fatty acid analysis) of single seeds from DHA-5C♯33_13 grown in either the field plots or glasshouse.

This study was conducted to compare two of the most widely used methods for the determination of the positional distribution of fatty acids within the triacylglycerol molecules from fats of animal origin. Method A involves incubation of the triacylglycerol fraction with an aqueous suspension of the lipase enzyme, separation of the reaction products...

1An alternative, sustainable source of omega‐3 long chain polyunsaturated fatty acids is widely recognized as desirable, helping to reduce pressure on current sources (wild capture fisheries) and providing a de novo source of these health beneficial fatty acids. This review will consider the efforts and progress to develop transgenic plants as terr...

Omega-3 fatty acids are characterized by a double bond at the third carbon atom from the end of the carbon chain. Latterly, long chain polyunsaturated omega-3 fatty acids such as eicosapentaenoic acid (EPA; 20:5Δ5,8,11,14,17) and docosahexanoic acid (DHA; 22:6 Δ4,7,10,13,16,19), which typically only enter the human diet via the consumption of oily...

Fats based on stearic acid could be a healthier alternative to existing oils especially hydrogenated fractions of oils or palm, but only a few non-tropical species produce oils with these characteristics. In this regard, newly developed high stearic oilseed crops could be a future source of fats and hard stocks rich in stearic and oleic fatty acids...

Total fatty acid composition of oil seeds from wild-type and transgenic C. sativa plants.

Comparison of C. sativa wild-type, RRes_EPA and RRes_DHA plants at 6 and 10 weeks old. Figure S2. Analysis of major phospholipid species in wild-type and engineered mature C.sativa seed. Figure S3. LC-MS/MS + MRM analysis of TAG from C. sativa engineered for production of EPA and/or DHA. Figure S4. Parallel analysis (GC-FID) of FAMEs derived from t...

Supplementary data

TLC screening of leaf tissue of Nicotiana tabacum T0 events at flowering stage as described in materials and methods. Lanes 42–44 are wild-type controls, which have relatively low TAG compared with the T0 transgenic event in lane 3 (described in this study). Figure S2 Southern blot of transgenic Nicotiana tabacum leaf DNA with NPTII probe demonstra...

An iterative approach to optimising the accumulation of non-native long chain polyunsaturated fatty acids in transgenic plants was undertaken in Arabidopsis thaliana. The contribution of a number of different transgene enzyme activities was systematically determined, as was the contribution of endogenous fatty acid metabolism. Successive iterations...

This article will focus on the modification of plant seed oils to enhance their nutritional composition. Such modifications will include C18 Δ6-desaturated fatty acids such as γ-linolenic and stearidonic acid, omega-6 long-chain polyunsaturated fatty acids such as arachidonic acid, as well as the omega-3 long-chain polyunsaturated fatty acids (ofte...

The synthesis and accumulation of long chain polyunsaturated fatty acids such as eicosapentaenoic acid has previously been demonstrated in the seeds of transgenic plants. However, the obtained levels are relatively low, indicating the need for further studies and the better definition of the interplay between endogenous lipid synthesis and the non-...

Omega-3 (ω-3) very long chain polyunsaturated fatty acids (VLC-PUFAs) such as eicosapentaenoic acid (EPA; 20:5 Δ5,8,11,14,17) and docosahexaenoic acid (DHA; 22:6 Δ4,7,10,13,16,19) have been shown to have significant roles in human health. Currently the primary dietary source of these fatty acids are marine fish; however, the increasing demand for f...

The role of acyl-CoA-dependent Δ6-desaturation in the heterologous synthesis of omega-3 long-chain polyunsaturated fatty acids was systematically evaluated in transgenic yeast and Arabidopsis thaliana. The acyl-CoA Δ6-desaturase from the picoalga Ostreococcus tauri and orthologous activities from mouse (Mus musculus) and salmon (Salmo salar) were s...

The present invention relates to the recombinant manufacture of polyunsaturated fatty acids. Specifically, it relates to acyltransferase polypeptides, polynucleotides encoding said acyltransferases as well as vectors, host cells, non-human transgenic organisms containing said polynucletides. Moreover, the present invention contemplates methods for...

The glycerol 3-phosphate acyltransferase (GPAT, EC 2.3.1.15) from sunflower (Helianthus annuus L.) microsomes has been characterised and partially purified. The in vitro determination of activity was optimized, and the maximum value for GPAT activity identified between 15 and 20 days after flowering. The apparent Michaelis-Menten K(m) for the glyce...

Enormous progress has been made in understanding genetic components required for the biosynthesis and modifica - tion of fatty acids in plants, thus facilitating the identification of sequences responsible for unusual fatty acid accumula - tion. Perhaps surprisingly, many early attempts to engineer the fatty acid composition of transgenic plants pr...

Dietary omega-3 polyunsaturated fatty acids have a proven role in reducing the risk of cardiovascular disease and precursor disease states such as metabolic syndrome. Although most studies have focussed on the predominant omega-3 fatty acids found in fish oils (eicosapentaenoic acid and docosahexaenoic acid), recent evidence suggests similar health...

Fatty acid desaturation in plastids and chloroplasts depends on the electron-donor activity of ferredoxins. Using degenerate oligonucleotides designed from known photosynthetic and heterotrophic plant ferredoxin sequences, two full-length ferredoxin cDNAs were cloned from sunflower (Helianthus annuus L.) leaves and developing seeds, HaFd1 and HaFd2...

The 1,3-random-2-random theory was proposed several years ago to explain the fatty acid distribution in vegetable oil triacylglycerols. However, by demonstrating an asymmetry between positions sn-1 and sn-3 in olive oil, cocoa butter, sunflower oil, etc., a number of studies have shown that this theory does not hold true for some oils and fatty aci...

The fatty acid distribution in triacylglycerols (TAGs) is a factor that contributes to the intrinsic properties of oils from different species variants. Many hypotheses have been proposed to explain the specific distribution of fatty acids in the different naturally occurring oils. Currently, the 1,3-random-2-random theory is more or less accepted,...

A method for plant tissue digestion and triacylglycerol (TAG) extraction followed by transmethylation of TAGs to produce the fatty acid methyl esters (FAMEs) from small storage tissue samples is presented. The method allows the analysis of both TAGs and FAMEs from the same sample. Several reagent mixtures and different experimental conditions were...

Triacylglycerols (TAG) are the main components of vegetable oils, accounting for almost 100% of the weight. Each oil has a characteristic pattern of TAG and the chemical, physical and nutritional properties of a particular oil are determined mainly by the abundance of its different TAG molecular species. Plant triacylglycerols are produced by the c...