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At least 55 records · Page 3

Fatty acid foams for nonselective physical removal of microplastics from aqueous solutions

Microplastics (MPs) are pervasive environmental contaminants whose removal from water remains a major challenge due to their small size, chemical diversity, and dynamic surface properties arising from environmental aging/weathering. Here, we present a concept of foam-based separation method that physically traps MPs in the foam phase using microtubular assemblies of 12-hydroxystearic acid. These foams are stabilized by anisotropic fatty acid microtubules formed in the presence of ethanolamine, which jam within the foam channels and suppress fluid drainage thereby enhancing MP retention and foam stability. MPs of different sizes, polymer compositions (including polystyrene, polypropylene, polyethylene terephthalate, and polytetrafluoroethylene), and weathered states were retained in the foam phase without requiring chemical modification or relying on chemical interactions between the fatty acid and MPs. Thermally induced transition of the fatty acid microtubules into nanomicelles above the characteristic phase transition temperature (∼35 °C) enables controlled foam collapse and recovery of trapped MPs. The cumulative removal efficiency can exceed 85% through multiple foaming cycles, matching predictions from a probabilistic retention model. This work shows that foams can provide a simple platform to trap MPs, thus providing a new physical-removal strategy that does not rely on the particles’ chemistry.

Guillot, Kennedy A. [Louisiana State Univ., Baton ↗

Identification of structural transitions in bacterial fatty acid binding proteins that permit ligand entry and exit at membranes

Fatty acid (FA) transfer proteins extract FA from membranes and sequester them to facilitate their movement through the cytosol. Detailed structural information is available for these soluble protein–FA complexes, but the structure of the protein conformation responsible for FA exchange at the membrane is unknown. Staphylococcus aureus FakB1 is a prototypical bacterial FA transfer protein that binds palmitate within a narrow, buried tunnel. Here, we define the conformational change from a “closed” FakB1 state to an “open” state that associates with the membrane and provides a path for entry and egress of the FA. Using NMR spectroscopy, we identified a conformationally flexible dynamic region in FakB1, and X-ray crystallography of FakB1 mutants captured the conformation of the open state. In addition, molecular dynamics simulations show that the new amphipathic α-helix formed in the open state inserts below the phosphate plane of the bilayer to create a diffusion channel for the hydrophobic FA tail to access the hydrocarbon core and place the carboxyl group at the phosphate layer. The membrane binding and catalytic properties of site-directed mutants were consistent with the proposed membrane docked structure predicted by our molecular dynamics simulations. Finally, the structure of the bilayer-associated conformation of FakB1 has local similarities with mammalian FA binding proteins and provides a conceptual framework for how these proteins interact with the membrane to create a diffusion channel from the FA location in the bilayer to the protein interior.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Plasticized terephthalate-based polyester blends containing fatty acids or esters thereof

A plasticized terephthalate-based polyester blend comprising a terephthalate-based polyester (e.g., polyethylene terephthalate, i.e., PET) homogeneously blended with a fatty acid or ester thereof, wherein said fatty acid or ester thereof is present in said polyester blend in an amount of 1-50 wt %. Also described herein is a method for producing a plasticized terephthalate-based polyester blend, the method comprising melt mixing a terephthalate-based polyester with a fatty acid or ester thereof at a temperature in a range of 230-250° C. to produce said polyester blend, wherein said fatty acid or ester thereof is present in said polyester blend in an amount of 1-50 wt % and is homogeneously blended with said terephthalate-based polyester.

Naskar, Amit K.↗

Using 14 C-acetate Pulse-chase Labeling to Study Fatty Acid and Glycerolipid Metabolism in Plant Leaves

Lipids metabolism is comprised of networks of reactions occurred in different subcellular compartments. Isotopic labeling is a good way to track the transformations and movements of metabolites without perturbing overall cellular metabolism. Fatty acids, the building blocks of membrane lipids and storage triacylglycerols, are synthesized in plastids. The immediate precursor for fatty acid synthesis is acetyl-CoA. Exogenous acetate is rapidly incorporated into fatty acids in leaves and isolated plastids because it can diffuse freely through cellular membranes, enter the plastid where it is rapidly metabolized to acetyl-CoA. Therefore, isotope-labeled acetate is often used as a tracer for the investigation of fatty acid synthesis and complex lipid metabolism in plants and other organisms. The basic principle of isotope labeling and its recent technical advances have been reviewed (Allen et al., 2015). The present protocol describes the use of 14C-labeled acetate to determine rates of fatty acid synthesis and degradation and to track the metabolism of glycerolipids in leaves. This method, which is often referred to as acetate pulse-chase labeling, has been widely used to probe various aspects of lipid metabolism (Allen et al., 2015), including the role of autophagy in membrane lipid turnover (Fan et al., 2019) and the interplay between lipid and starch metabolism pathways (Yu et al., 2018).

14C-acetate pulse-chase↗

Fuel property evaluation of unique fatty acid methyl esters containing β-hydroxy esters from engineered microorganisms

Unique fatty acid methyl esters (FAME) containing ..beta..-hydroxy esters were produced using an engineered microorganism by glucose fermentation. This study investigated the properties of the unique FAME mixture both neat and in blends with conventional diesel, as well as properties of ..beta..-hydroxy esters. The unique FAME blend contained relatively shorter-chain FAME (average fatty acid chain carbon number 14.6) with 58 % monounsaturated fatty acids and 9 % saturated and monounsaturated ..beta..-hydroxy acid chains. The unique FAME had significantly lower distillation T90 (321 °C versus 352 °C) and higher cetane number (56.7 versus 52) compared to soy biodiesel. Cloud points were within method repeatability. Unexpectedly (because of the lack of methylene-interrupted double bonds), the unique FAME had low oxidation stability (1.5 h) as determined by Rancimat induction period. Stability could be improved through addition of commonly used antioxidants. We speculate that monounsaturated ..beta..-hydroxy FAME may be the source of this instability. Blends with conventional diesel up to 50 vol% showed similar kinematic viscosity (within method repeatability) as blends of conventional FAME. The unique FAME had no effect on distillation T90 even at the 80% blend level. A 30 vol% blend into conventional diesel had a Rancimat induction period of only 2 h, very nearly the same as the neat unique FAME sample. The addition of antioxidants produced blends of acceptable stability. Based on an assessment of the properties of individual ..beta..-hydroxy FAME molecules, they have higher boiling point, higher cloud point, lower cetane number, and potentially lower storage stability than analogous FAME not having the ..beta..-hydroxy group. Removing them from the fuel product in the production process may result in a biodiesel product with superior properties to what is on the market today.

09 BIOMASS FUELS↗

Genomic Analysis of the Natural Variation of Fatty Acid Composition in Seed Oils of Camelina sativa

Camelina sativa is an oilseed crop that has shown strong promise as a biofuel feedstock. The profile of fatty acids greatly influences the oil quality; however, genetic mechanisms that determine the natural variation of fatty acid composition in camelina are not fully understood. A genome wide association study (GWAS) was performed to uncover genetic loci that may contribute to the contents of major fatty acids such as oleic and linolenic acids in camelina seed. Two approaches were taken to improve the GWAS efficiency. First, growing a diversity panel of 212 accessions in four locations and two nitrogen fertilization conditions revealed great variation in fatty acid contents in seeds. Second, using an improved reference genome, abundant markers, including 203,320 single nucleotide polymorphisms (SNPs) and 99,067 insertions/deletions (indels), were developed, which refined the population structure of the diversity panel. GWAS resulted in 118 genetic markers across 31 trait/treatment conditions. Closely linked markers were determined based on linkage decay and by comparing secondarily associated markers when highly associated ones were removed. Candidate genes were examined by comparing the pangenomes of 12 high-quality reference genomes. This study provides new resources to understand seed lipid metabolism and improve camelina oils through molecular breeding.

Life Sciences & Biomedicine - Other Topics↗

QTL Mapping of Seed Fatty Acid Contents in Camelina sativa Under Heat Stress

Heat stress alters oil quality in oilseed crops, yet its genetic underpinnings in Camelina sativa remain unclear. This study investigated the genetic basis of heat-induced changes in seed fatty acids using a recombinant inbred line (RIL) population derived from a cross between two camelina varieties, Suneson and Pryzeth. Exposure to high temperature during reproductive growth led to increased proportions of saturated (C16:0, C18:0) and monounsaturated (C18:1) fatty acids, whereas polyunsaturated C18:3, total unsaturated fatty acids (UFA) and the PUFA/MUFA ratio were decreased, suggesting an inhibition of the C18:1 → C18:2 → C18:3 desaturation pathway. A high-density linkage map (4981 bins across 20 chromosomes) was built, and 25 QTLs for fatty acids were detected, with hotspots on chromosomes 1, 9, 12, 13, 16, and 20. A major QTL on chromosome 1 (~ 80 cM) explained the largest variance component for PUFA/MUFA under heat. Three desaturase genes (FAD2, FAD7, FAD8) were located within key QTL intervals, nominating them as candidates for modulating unsaturation under elevated temperature. These results provide a genetic basis for fine mapping and functional validation, supporting future molecular and breeding efforts to stabilize oil quality under warming conditions.

Camelina↗

Target of 1 Rapamycin kinase is a positive regulator of plant fatty acid 2 and lipid synthesis

In eukaryotes, Target of Rapamycin (TOR), a conserved protein sensor kinase, integrates a diverse set of environmental cues, including growth factor signals, energy availability, and nutritional status, to direct cell growth. In plants, TOR is activated by light and sugars and regulates a wide range of cellular processes, including protein synthesis and metabolism. Fatty acid synthesis is key to membrane biogenesis that in turn, is required for cell growth. To elucidate the primary regulatory role(s) of TOR in lipid metabolism, we followed fatty acid and lipid changes in plants with altered TOR protein levels or activity for short durations, using Nicotiana benthamiana leaves, Arabidopsis seedlings and Brassica napus cell suspension cultures. Transient expression of TOR significantly elevated the levels of total fatty acids in Nicotiana benthamiana leaves, while treatment of Arabidopsis seedlings with Torin 2, a TOR specific inhibitor, for one day, caused significant reductions in fatty acids and membrane lipids. Similarly, incubating oil-producing Brassica napus suspension culture cells with Torin 2 for eight hours led to significant decreases in the levels of TFA and TAG. Taken together the results from three independent systems presented here establishes that TOR positively regulates lipid synthesis in plants, consistent with its role in animals. Furthermore, RNA-seq analysis of Torin 2-treated Arabidopsis seedlings showed that TOR promotes the upregulation of a number of genes involved in de novo fatty acid synthesis while downregulating genes involved in lipid turnover, which we propose as a mechanistic explanation for its promotion of lipid synthesis and accumulation.

59 BASIC BIOLOGICAL SCIENCES↗

Fatty Acid and Alcohol Metabolism in Pseudomonas putida: Functional Analysis Using Random Barcode Transposon Sequencing.

With its ability to catabolize a wide variety of carbon sources and a growing engineering toolkit, Pseudomonas putida KT2440 is emerging as an important chassis organism for metabolic engineering. Despite advances in our understanding of the organism, many gaps remain in our knowledge of the genetic basis of its metabolic capabilities. The gaps are particularly noticeable in our understanding of both fatty acid and alcohol catabolism, where many paralogs putatively coding for similar enzymes coexist, making biochemical assignment via sequence homology difficult. To rapidly assign function to the enzymes responsible for these metabolisms, we leveraged random barcode transposon sequencing (RB-Tn-Seq). Global fitness analyses of transposon libraries grown on 13 fatty acids and 10 alcohols produced strong phenotypes for hundreds of genes. Fitness data from mutant pools grown on fatty acids of varying chain lengths indicated specific enzyme substrate preferences and enabled us to hypothesize that DUF1302/DUF1329 family proteins potentially function as esterases. From the data, we also postulate catabolic routes for the two biogasoline molecules isoprenol and isopentanol, which are catabolized via leucine metabolism after initial oxidation and activation with coenzyme A (CoA). Because fatty acids and alcohols may serve as both feedstocks and final products of metabolic-engineering efforts, the fitness data presented here will help guide future genomic modifications toward higher titers, rates, and yields. IMPORTANCE To engineer novel metabolic pathways into P. putida , a comprehensive understanding of the genetic basis of its versatile metabolism is essential. Here, we provide functional evidence for the putative roles of hundreds of genes involved in the fatty acid and alcohol metabolism of the bacterium. These data provide a framework facilitating precise genetic changes to prevent product degradation and to channel the flux of specific pathway intermediates as desired.

59 BASIC BIOLOGICAL SCIENCES↗

Persistent fatty acid catabolism during plant oil synthesis

Plant lipids are an essential energy source for diets and are a sustainable alternative to petroleum-based fuels and feedstocks. Fatty acid breakdown during seed germination is crucial for seedling establishment but unexpected during seed filling. Here, we demonstrate that the simultaneous biosynthesis and degradation of fatty acids begins early and continues across all phases of oil filling and throughout the photoperiod. Tests in camelina, rapeseed, and an engineered high-oil tobacco line confirmed that concomitant synthesis and breakdown in oil-producing tissues over development is the rule rather than the exception. Furthermore, we show that transgenics, designed to elevate fatty acid biosynthesis, failed to achieve anticipated increases in storage lipid levels due to increased degradation, potentially explaining the underperformance of engineered lines compared to expectations more generally.

coenzyme A↗

Reduction of Chemokine CXCL9 Expression by Omega-3 Fatty Acids via ADP-Ribosylhydrolase ARH3 in MIN6 Insulin-Producing Cells

Type 1 diabetes (T1D) results from the autoimmune destruction of the insulin-producing β cells of the pancreas. Omega-3 fatty acids protect β cells and reduce the incidence of T1D, but the mechanism is poorly understood. We have shown that omega-3 fatty acids reduce pro-inflammatory cytokine-mediated β-cell apoptosis by upregulating the expression of the ADP-ribosylhydrolase ARH3. Here, we further investigate the β-cell protection mechanism of ARH3 by performing siRNA analysis of its gene Adprhl2 in MIN6 insulin-producing cells, subsequent treatment with a cocktail of the pro-inflammatory cytokines IL-1β + IFN-γ + TNF-α, followed by proteomics analysis. ARH3 regulated proteins from several pathways related to the nucleus (splicing, RNA surveillance, and nucleocytoplasmic transport), mitochondria (metabolic pathways), and endoplasmic reticulum (protein folding). ARH3 also regulated the levels of proteins related to antigen processing and presentation, and the chemokine-signaling pathway. We further studied the role of ARH3 in regulating the chemokine CXCL9. We found that ARH3 reduces the cytokine-induced expression of CXCL9, which is dependent on omega-3 fatty acids. In conclusion, we demonstrate that omega-3 fatty acids regulate CXCL9 expression via ARH3, which may have a role in protecting β cells from immune attack thereby preventing T1D development.

59 BASIC BIOLOGICAL SCIENCES↗

Genetic and Biochemical Investigation of Seed Fatty Acid Accumulation in Arabidopsis

As a vegetable oil, consisting principally of triacylglycerols, is the major storage form of photosynthetically-fixed carbon in oilseeds which are of significant agricultural and industrial value. Photosynthesis in chlorophyll-containing green seeds, along with photosynthesis in leaves and other green organs, generates ATP and reductant (NADPH and NADH) needed for seed fatty acid production. However, contribution of seed photosynthesis to fatty acid accumulation in seeds have not been well-defined. Here, we report the contribution of seed-photosynthesis to fatty acid production by probing segregating green (photosynthetically-competent) and non-green or yellow (photosynthetically-non-competent) seeds in siliques of an Arabidopsis chlorophyll synthase mutant. Using this mutant, we found that yellow seeds lacking photosynthetic capacity reached 80% of amounts of oil in green seeds at maturity. Combining this with studies using shaded siliques, we determined that seed-photosynthesis accounts for 20% and silique and leaf/stem photosynthesis each account for ~40% of the ATP and reductant for seed oil production. Transmission electron microscopy (TEM) and pyridine nucleotides and ATP analyses revealed that seed photosynthesis provides ATP and reductant for oil production mostly during early development, as evidenced by delayed oil accumulation in non-green seeds. Transcriptomic analyses suggests that the oxidative pentose phosphate pathway could be the source of carbon, energy and reductants required for fatty acid synthesis beyond the early stages of seed development.

Nwafor, Chinedu Charles↗

Higher-order interaction effects among operating conditions and feedstocks shape reactor microbiomes and fatty acid production profiles

Arrested anaerobic digestion (AAD) offers a promising route for producing fatty acids (FAs) from organic residues, yet optimal conditions for selectively generating medium-chain fatty acids (MCFAs) remain poorly defined. Here, we systematically evaluated the main and interaction effects of pH (5, 7, 9), feedstock (food waste, manure), temperature (35 and 45 °C), and inoculum source on microbiome composition and FA production. Anaerobic digester sludge and a novel bison rumen inoculum were compared. Significant higher-order interactions among operating parameters governed FA profiles and microbiome structure. Butyric acid production was driven by a three-way interaction among pH, feedstock, and temperature (p < 0.001), with maximum concentrations achieved in food waste reactors at pH 5.0 and 35 °C (1.2 ± 0.1 g L −1 with sludge and 1.1 ± 0.3 g L −1 with rumen). MCFA production exhibited significant four-way interactions (p < 0.1 to p < 0.001). At 45 °C and pH 5.0, inoculum source tuned MCFA selectivity: sludge favored pentanoic acid (0.4 ± 0.1 g L −1 ), whereas rumen favored hexanoic and heptanoic acids (up to 0.4 ± 0.2 g L −1 ). Manure reactors produced < 0.2 g L −1 MCFAs under all conditions. Genera, including Megasphaera, Prevotella, and Lactobacillus, were associated with production of specific MCFAs. PICRUSt2-based pathway predictions were consistent with MCFA production patterns and suggested a potential role for lactic acid–driven chain elongation pathways. This study provides insights into how interacting operating conditions shape AAD microbiomes, their FA profiles, and advances the trajectory of research aimed at engineering robust and controllable microbiomes for waste valorization.

09 BIOMASS FUELS↗

A 2-component blend of coconut oil-derived fatty acids as an oviposition deterrent against Drosophila suzukii (Drosophilidae: Diptera)

Abstract Coconut free fatty acid (CFFA), a mixture of 8 fatty acids derived from coconut oil, is an effective repellent and deterrent against a broad array of hematophagous insects. In this study, we evaluated the oviposition deterrent activity of CFFA on spotted-wing drosophila (SWD; Drosophila suzukii), a destructive invasive pest of berries and cherries, and identified bioactive key-deterrent compounds. In laboratory 2-choice tests, CFFA deterred SWD oviposition in a dose-dependent manner with the greatest reduction (99%) observed at a 20-mg dose compared with solvent control. In a field test, raspberries treated with 20-mg CFFA received 64% fewer SWD eggs than raspberries treated with the solvent control. In subsequent laboratory bioassays, 2 of CFFA components, caprylic and capric acids, significantly reduced SWD oviposition by themselves, while 6 other components had no effect. In choice and no-choice assays, we found that a blend of caprylic acid and capric acid, at equivalent concentrations and ratio as in CFFA, was as effective as CFFA, while caprylic acid or capric acid individually were not as effective as the 2-component blend or CFFA at equivalent concentrations, indicating the 2 compounds as the key oviposition deterrent components for SWD. The blend was also as effective as CFFA for other nontarget drosophilid species in the field. Given that CFFA compounds are generally regarded as safe for humans, CFFA and its bioactive components have potential application in sustainably reducing SWD damage in commercial fruit operations, thereby reducing the sole reliance on insecticides.

Entomology↗

Do betaine lipids replace phosphatidylcholine as fatty acid editing hubs in microalgae?

Acyl editing refers to a deacylation and reacylation cycle on a lipid, which allows for fatty acid desaturation and modification prior to being removed and incorporated into other pools. Acyl editing is an important determinant of glycerolipid synthesis and has been well-characterized in land plants, thus this review begins with an overview of acyl editing in plants. Much less is known about acyl editing in algae, including the extent to which acyl editing impacts lipid synthesis and on which lipid substrate(s) it occurs. This review compares what is known about acyl editing on its major hub phosphatidylcholine (PC) in land plants with the evidence for acyl editing of betaine lipids such as diacylglyceryltrimethylhomoserine (DGTS), the structural analog that replaces PC in several species of microalgae. In land plants, PC is also known to be a major source of fatty acids and diacylglycerol (DAG) for synthesis of the neutral lipid triacylglycerol (TAG). We review the evidence that DGTS contributes substantially to TAG accumulation in algae as a source of fatty acids, but not as a precursor to DAG. We conclude with evidence of acyl editing on other membrane lipid substrates in plants and algae apart from PC or DGTS, and discuss future analyses to elucidate the role of DGTS and other betaine lipids in acyl editing in microalgae.

59 BASIC BIOLOGICAL SCIENCES↗

Solvent‐free enzymatic esterification of free fatty acids with glycerol for biodiesel application: Optimized using the Taguchi experimental method

Abstract Presence of free fatty acids along with glycerides poses a technical difficulty for biodiesel production. This work used a Taguchi L9 design to optimize the solvent‐free enzymatic process to result in the esterification of oleic acid with glycerol. Under optimal conditions the esterification reaction temperature of 60°C, enzyme dose of 5 wt%, glycerol: oleic acid molar ratio of 5:1, and reaction time of 3 h, a 75.235 ± 2.19% conversion of oleic acid to esters was achieved. With the addition of molecular sieves, the conversion increased to 86.73% ± 1.09%. However, using the parameters predicted by Taguchi design (60°C, 5 wt%, 5:1, and 4.5 h), 88.5% ± 1.11% of oleic acid could be converted to esters derivative. Diglycerides were the major product, and the reaction equilibrium was attained after 4 h. The immobilized enzyme could be used up to seven times with only a 10% reduction in the conversion. Thus, the process can efficiently reduce the free fatty acid content of oil to make it suitable for biodiesel production.

09 BIOMASS FUELS↗

Genetically-determined variations in photosynthesis indicate roles for specific fatty acid species in chilling responses

Using a population of recombinant inbred lines (RILs) cowpea (Vigna unguiculata. L. Walp), we tested for co-linkages between lipid contents and chilling responses of photosynthesis. Under low-temperature conditions (19°C/13°C, day/night), we observed co-linkages between quantitative trait loci intervals for photosynthetic light reactions and specific fatty acids, most strikingly, the thylakoid-specific fatty acid 16:1 Δ3trans found exclusively in phosphatidylglycerol (PG 16:1t). By contrast, we did not observe co-associations with bulk polyunsaturated fatty acids or high-melting-point-PG (sum of PG 16:0, PG 18:0 and PG 16:1t) previously thought to be involved in chilling sensitivity. These results suggest that in cowpea, chilling sensitivity is modulated by specific lipid interactions rather than bulk properties. We were able to recapitulate the predicted impact of PG 16:1t levels on photosynthetic responses at low temperature using mutants and transgenic Arabidopsis lines. Because PG 16:1t synthesis requires the activity of peroxiredoxin-Q, which is activated by H 2 O 2 and known to be involved in redox signalling, here we hypothesise that the accumulation of PG 16:1t occurs as a result of upstream effects on photosynthesis that alter redox status and production of reactive oxygen species.

59 BASIC BIOLOGICAL SCIENCES↗

Aureochromes maintain polyunsaturated fatty acid content in Nannochloropsis oceanica

Abstract Nannochloropsis oceanica, like other stramenopile microalgae, is rich in long-chain polyunsaturated fatty acids (LC-PUFAs) such as eicosapentaenoic acid (EPA). We observed that fatty acid desaturases (FADs) involved in LC-PUFA biosynthesis were among the strongest blue light-induced genes in N. oceanica CCMP1779. Blue light was also necessary for maintaining LC-PUFA levels in CCMP1779 cells, and growth under red light led to a reduction in EPA content. Aureochromes are stramenopile-specific proteins that contain a light–oxygen–voltage (LOV)-sensing domain that associates with a flavin mononucleotide and is able to sense blue light. These proteins also contain a basic leucine zipper DNA-binding motif and can act as blue light-regulated transcription factors by associating with an E-box like motif, which we found enriched in the promoters of blue light-induced genes. We demonstrated that, in vitro, two CCMP1779 aureochromes were able to absorb blue light. Moreover, the loss or reduction of the expression of any of the three aureochrome genes led to a decrease in the blue light-specific induction of several FADs in CCMP1779. EPA content was also significantly reduced in NoAUREO2 and NoAUREO4 mutants. Taken together, our results indicate that aureochromes mediate blue light-dependent regulation of LC-PUFA content in N. oceanica CCMP1779 cells.

Plant Sciences↗