Lipid Extracts Research Articles

Nanodisc assembly from bacterial total lipid extracts

Nanodiscs are discoidal lipoproteins that have often been used as vehicles to study membrane proteins in their native configuration. Nanodiscs have been primarily made from synthetic lipids. However, nanodiscs also offer a format by which native lipids can be studied in their natural configuration. Here, we present a method to synthesize nanodiscs from bacterial total lipid extracts using the biothreat agent, Yersinia pestis, as a proof-of-concept. The creation of nanoparticles entirely composed of bacterial lipids supports membrane characterization and vaccine antigen discovery without the inherent safety concerns associated with live bacterial cells of this Tier 1 select agent pathogen.

Effects of pre-treatment, historical age, and sample characteristics on the stable isotope analyses of killer whale (Orcinus orca) bone.

Stable isotope analysis of bone provides insight into animal foraging and allows for ecological reconstructions over time, however pre-treatment is required to isolate collagen. Pre-treatments typically consist of demineralization to remove inorganic components and/or lipid extraction to remove fats, but these protocols can differentially affect stable carbon (δ13C) and nitrogen (δ15N) isotope values depending on the chemicals, tissues, and/or species involved. Species-specific methodologies create a standard for comparability across studies and enhance understanding of collagen isolation from modern cetacean bone. Elemental analyzers coupled to isotope ratio mass spectrometers were used to measure the δ13C and δ15N values of powdered killer whale (Orcinus orca) bone that was intact (control) or subjected to one of three experimental conditions: demineralized, lipid-extracted, and both demineralized and lipid-extracted. Additionally, C:N ratios were evaluated as a proxy for collagen purity. Lastly, correlations were examined between control C:N ratios vs. historical age and control C:N ratios vs. sample characteristics. No significant differences in the δ15N values were observed for any of the experimental protocols. However, the δ13C values were significantly increased by all three experimental protocols: demineralization, lipid extraction, and both treatments combined. The most influential protocol was both demineralization and lipid extraction. Measures of the C:N ratios were also significantly lowered by demineralization and both treatments combined, indicating the material was closer to pure collagen after the treatments. Collagen purity as indicated via C:N ratio was not correlated with historical age nor sample characteristics. If only the δ15N values from killer whale bone are of interest for analysis, no pre-treatment seems necessary. If the δ13C values are of interest, samples should be both demineralized and lipid-extracted. As historical age and specimen characteristics are not correlated with sample contamination, all samples can be treated equally.

Comprehensive comparison of fatty acid methyl ester profile in different food matrices using microwave-assisted extraction and derivatization methods and comprehensive two-dimensional gas chromatography coupled with flame ionization detection

Analyzing fatty acids provides key insights into fat composition for industrial applications and their implications for nutrition and health. Typically, fatty acid analysis involves extracting lipids from the matrix and converting them into fatty acid methyl esters (FAME) through a derivatization process before gas chromatography (GC) analysis. Either one-step or two-step procedures can be found in the literature and as official methods. In this work, different methods exploiting microwave-assisted processes were compared with two official methods from the American Oil Chemical Society (AOCS). Especially, two types of microwave-assisted extractions were employed: solvent extraction and extraction with hydrolysis. The extracts were derivatized using either BF3 or a microwave-assisted methanolic hydrogen chloride solution. These combinations of extraction and derivatization methods were compared also with one-step microwave-assisted extraction and derivatization, and two AOCS reference methods, resulting in seven different methods applied to six different food matrices. The performance of the different procedures was compared based on the FAME profile obtained from the comprehensive two-dimensional GC (GC × GC)-FID analysis.Microwave-assisted processes were shown to be effective, yielding results comparable to the official methods in both the one-step and two-step methods. Moreover, it was shown that the BF3 derivatization could be safely replaced with microwave-assisted derivatization with methanolic hydrogen chloride, providing equivalent performances while enhancing operator safety and environmental friendliness. Some discrepancies in the FAMEs profile were highlighted for the sample of oats, the only explicitly requiring acidic hydrolysis for lipid extraction. Further studies are required to understand the reasons behind these differences and develop a suitable modified method. In conclusion, all the methods were evaluated for greenness and blueness with two specific tools: AGREEprep and BAGI.

A novel enzyme-assisted n-hexane/ethanol biphasic method to extract and separate heterotrophic Chlorella lipids for anti-inflammatory activity

Chlorella lipids are rich in essential fatty acids and have potential for development in food and healthcare products. This study established a novel enzyme-assisted biphasic extraction to obtain functional lipids from Chlorella sp. MBFJNU-17 biomass, and evaluated the anti-inflammatory function of the extracted lipids. The obtained results showed that the enzyme-assisted biphasic extraction mediated with pectinase and organic solvents (n-hexane and ethanol) achieved 17.3 % Chlorella cell wall disruption and 268.30 mg/g lipid extraction rate. After the optimization of this process, the n-hexane phase had 83.20 % neutral lipids (NLs), the ethanol phase contained 94.02 % polar lipids (53.65 % glycolipids (GLs) and 40.37 % phospholipids (PLs)), manifesting that the established biphasic lipid extraction exhibited the attracting feature to separate the NLs and polar lipids after lipids extraction. In term of the anti-inflammatory, the Chlorella polar lipids were found to perform superior anti-inflammatory effect with lower levels of inflammatory factors TNF-α and IL-6 than those of NLs. Compared to the lipids in the n-hexane and ethanol phases, three individual lipids (NL, GLs and PLs) performed poorer anti-inflammatory effects, indicating that the synergistic action could be effectively improved by the combination of three lipids. Taken together, the developed enzyme-assisted biphasic extraction method for Chlorella sp. MBFJNU-17 lipids was the potential lipid extraction process to concurrently extract and separate anti-inflammatory lipids. The study offered essential and scientific details to exploit high-value Chlorella lipids for nutritional and healthy bioproducts.

Microemulsions strongly promoted the activity of α-bisabolol against different Leishmania species and its skin permeation

This study aimed to develop microemulsions (MEs) containing α-bisabolol for the topical treatment of cutaneous leishmaniasis (CL). Initially, pseudoternary phase diagrams were developed using α-bisabolol as the oil phase, Eumulgin® CO 40 as the surfactant, Polymol® HE as the co-surfactant, and distilled water as the aqueous phase. Two transparent liquid systems (TLS) containing 5% of α-bisabolol were selected and characterized (F5E25 and F5EP25). Next, skin permeation and retention assays were performed using Franz cells. The interaction of the formulation with the stratum corneum (SC) was evaluated using the FTIR technique. The cytotoxicity was evaluated in murine peritoneal macrophages. Finally, the antileishmanial activity of microemulsions was determined in promastigotes and amastigotes of L. amazonensis (strain MHOM/BR/77/LTB 0016). As a result, the selected formulations showed isotropy, nanometric size (below 25 nm), Newtonian behavior and pH ranging from 6.5 to 6.9. The MEs achieved a 2.5-fold increase in the flux and skin-permeated amount of α-bisabolol. ATR-FTIR results showed that microemulsions promoted fluidization and extraction of lipids and proteins of the stratum corneum, increasing the diffusion coefficient and partition coefficient of the drug in the skin. Additionally, F5E25 and F5EP25 showed higher activity against promastigotes (IC50 13.27 and 18.29, respectively) compared to unencapsulated α-bisabolol (IC50 53.8). Furthermore, F5E25 and F5EP25 also showed antileishmanial activity against intracellular amastigotes of L. amazonensis, with IC50 50 times lower than free α-bisabolol and high selectivity index (up to 15). Therefore, the systems obtained are favorable to topical administration, with significant antileishmanial activity against L. amazonensis promastigotes and amastigotes, being a promising system for future in vivo trials.

Fueling the future with fungi: Efficient lipid extraction from Ganoderma Mycelium Biomass

Ganoderma lucidum mycelial biomass produced via submerged-liquid fermentation was subjected to different lipid extraction methods; soxhlet extraction (SXE), solvent extraction (SVE) and ultrasonic-assisted extraction (UAE), followed by a comparison in its lipid yield. The experiment was carried out using 5 g of biomass material, with 50-300 ml hexane solvent, and the temperature was set at 60 ° C for 1-9 hours. It was observed that the lipid yield was found to be influenced by the extraction time and solvent quantity. SVE was observed to be more effective for lipid production of 20.36 % (2 h and 100 ml hexane) in comparison with SXE (18.8 %) and UAE (7.50 %). In general, G. lucidum mycelial biomass may be used as a potential raw material and cost-effective solution for an alternative biodiesel production process in the future. Hence, future research work should focus on other novel extraction techniques as well as optimizing the other extraction parameters in improving the lipid yield from G. lucidum mycelial biomass.

Defining the functional properties of cyclopropane fatty acid synthase from Pseudomonas aeruginosa PAO1

Cyclopropane fatty acid synthases (CFAS) catalyze the conversion of unsaturated fatty acids to cyclopropane fatty acids (CFAs) within bacterial membranes. This modification alters the biophysical properties of membranes and has been correlated with virulence in several human pathogens. Despite the central role played by CFAS enzymes in regulating bacterial stress responses, the mechanistic properties of the CFAS enzyme family and the consequences of CFA biosynthesis remain largely uncharacterized in most bacteria. We report the first characterization of the CFAS enzyme from Pseudomonas aeruginosa (PA) – an opportunistic human pathogen with complex membrane biology that is frequently associated with antimicrobial resistance and high tolerance to various external stressors. We demonstrate that CFAs are produced by a single enzyme in PA and that cfas gene expression is upregulated during the transition to stationary phase and in response to oxidative stress. Analysis of PA lipid extracts reveal a massive increase in CFA production as PA cells enter stationary phase and help define the optimal membrane composition for in vitro assays. The purified PA-CFAS enzyme forms a stable homodimer and preferentially modifies phosphatidylglycerol lipid substrates and membranes with a higher content of unsaturated acyl chains. Bioinformatic analysis across bacterial phyla shows highly divergent amino acid sequences within the lipid binding domain of CFAS enzymes, perhaps suggesting distinct membrane binding properties among different orthologues. This work lays an important foundation for further characterization of CFAS in Pseudomonas aeruginosa and for examining the functional differences between CFAS enzymes from different bacteria.

Effective Lipid Extraction from Fat Balls Using Liquefied Dimethyl Ether: Process Optimization with a Box-Behnken Design.

In recent years, lipids reused from urban wastewater materials have come to prominence as possible raw materials for biodiesel production. The present work investigated liquefied dimethyl ether (DME) for the lipid extraction of fat balls from sewage pumping stations. A response surface methodology (RSM) based on a Box-Behnken design (BBD) was utilized to optimize DME extraction parameters (sample size, velocity of liquefied DME, and DME/sample ratio). The maximum lipid yield was 65.2% under optimal DME extraction conditions (sample size 1 mm, velocity of liquefied DME 3.3 m/h, and DME/sample ratio 80 mL/g). Under the optimum conditions, the DME technique exhibited higher lipid recovery than that of mechanical shaking extraction (49.0%) or Soxhlet extraction (62.0%). The extracted lipids were converted into biodiesel, resulting in an approximately 35.2-46.2% biodiesel yield. Furthermore, the fatty acid methyl ester composition of the extracted lipids was characterized. These significant findings highlight the promising potential of fat balls as sustainable biodiesel feedstocks and provide valuable insight that will aid the development of better technology for lipid extraction.

Avocado seed extract encapsulated in zein nanoparticles as a functional ingredient

Nano-delivery of natural compounds has gained increasing attention in the food and pharmaceutical ingredients industry. Avocado seed was extracted by a total lipid extraction and the presence of two acetogenin compounds was confirmed by the high-performance liquid chromatography (LC-MS) technique. Then, the avocado seed extract (Avo) was encapsulated into zein nanoparticles by liquid-liquid dispersion and the formation of zein nanoparticles loaded with avocado seed extract (Avo-ZNPs) was confirmed by FTIR analysis. The Avo-ZNPs with various Avo contents showed an average size ranging from 166.9 ± 14.1 to 305.2 ± 16.2 nm with 0.19–0.26 of polydispersity index (PDI), and a surface charge range was −59.5 ± 8.1 to −32.9 ± 0.4 mV, as measured by dynamic light scattering (DLS). The entrapment efficiency (EE) values were up to 82 % and the highest loading capacity (LoC) was 14.8 %. The Avo-ZNPs were found to have significantly higher antioxidant potency compared to the control, as measured by DPPH and FRAP assays. Moreover, the zein nanoparticles were also biocompatible with the acute myeloid leukemia (AML) cell lines, and encapsulated avocado extract induced leukemia cell death. Thus, zein nanoparticles could be a promising delivery system for natural avocado seed extract to be used in the medical and food industries.

Zero-waste biorefining co-products from ultrasonically assisted deep eutectic solvent-pretreated Chlorella biomass: Sustainable production of biodiesel and bio-fertilizer

Microalgal biomass is gaining increasing attention to produce high-value co-products. This study proposes integrating Chlorella microalgal biomass into a zero-waste biorefining system, aiming to produce biodiesel and biofertilizer. It investigates optimal conditions for ultrasound-assisted deep eutectic solvent (DES) pretreatment and lipid recovery to enhance the extraction of lipids. Optimal DES pretreatment was identified as a 1.6:1 acetic acid-to-choline chloride molar ratio, 0.36 g biomass loading, and 2.50 min of pretreatment. Lipid recovery succeeded with a 10-minute extraction time and a 1:3 methanol-to-butanol volume ratio. These conditions yielded biodiesel-quality lipids at 139.52 mg/g microalgal biomass with superior fuel characteristics. The de-oiled microalgal biomass residue exhibited promise as a lettuce biofertilizer, enhancing photosynthetic pigments but potentially reducing yields by 40 %. The study also notes changes in rhizosphere microbial communities, indicating both stimulatory and inhibitory effects on beneficial microbes. This study has the potential to enhance sustainability in energy, agriculture, and the environment.

Nanoscale Perturbations of Lipid Bilayers Induced by Magainin 2: Insights from AFM Imaging and Force Spectroscopy

This study explores the impact of the antimicrobial peptide magainin 2 (Mag2) on lipid bilayers with varying compositions. We employed high-resolution atomic force microscopy (AFM) to reveal a dynamic spectrum of structural changes induced by Mag2. Our AFM imaging unveiled distinct structural alterations in zwitterionic POPC bilayers upon Mag2 exposure, notably the formation of nanoscale depressions within the bilayer surface, which we term as "surface pores" to differentiate them from transmembrane pores. These surface pores are characterized by a limited depth that does not appear to fully traverse the bilayer and reach the opposing leaflet. Additionally, our AFM-based force spectroscopy investigation on POPC bilayers revealed a reduction in bilayer puncture force (FP) and Young's modulus (E) upon Mag2 interaction, indicating a weakening of bilayer stability and increased flexibility, which may facilitate peptide insertion. The inclusion of anionic POPG into POPC bilayers elucidated its modulatory effects on Mag2 activity, highlighting the role of lipid composition in peptide-bilayer interactions. In contrast to surface pores, Mag2 treatment of E. coli total lipid extract bilayers resulted in increased surface roughness, which we describe as a fluctuation-like morphology. We speculate that the weaker cohesive interactions between heterogeneous lipids in E. coli bilayers may render them more susceptible to Mag2-induced perturbations. This could lead to widespread disruptions manifested as surface fluctuations throughout the bilayer, rather than the formation of well-defined pores. Together, our findings of nanoscale bilayer perturbations provide useful insights into the molecular mechanisms governing Mag2-membrane interactions.

A Review of Grease Trap Waste Management in the US and the Upcycle as Feedstocks for Alternative Diesel Fuels

As byproducts generated by commercial and domestic food-related processes, FOGs (fats, oils, and grease) are the leading cause of sewer pipe blockages in the US and around the world. Grease trap waste (GTW) is a subcategory of FOG currently disposed of as waste, resulting in an economic burden for GTW generators and handlers. This presents a global need for both resource conservation and carbon footprint reduction, particularly through increased waste upcycling. Therefore, it is critical to better understand current GTW handling practices in the context of the urban food–energy–water cycle. This can be accomplished with firsthand data collection, such as onsite visits, phone discussions, and targeted questionnaires. GTW disposal methods were found to be regional and correspond to key geographical locations, with landfill operations mostly practiced in the Midwest regions, incineration mainly in the Northeast and Mid-Atlantic regions, and digestion mainly in the West of the US. Select GTW samples were analyzed to evaluate their potential reuse as low-cost feedstocks for biodiesel or renewable diesel, which are alternatives to petroleum diesel fuels. Various GTW lipid extraction technologies have been reviewed, and more studies were found on converting GTW into biodiesel rather than renewable diesel. The challenges for these two pathways are the high sulfur content in biodiesel and the metal contents in renewable diesel, respectively. GTW lipid extraction technologies should overcome these issues while producing minimum-viable products with higher market values.

Lipid extract of microalgae Chlorella vulgaris and Arthrospira platensis as a bioactive ingredient.

A growing market for seaweed products has emerged in the last 20 years, with microalgae and their extracts representing a source of high-value chemicals. In this study, antioxidative and anti-inflammatory activities of lipid fraction extract from Chlorella vulgaris (Chlorella) and Arthrospira platensis (Spirulina) on Normal Human Dermal Fibroblasts (NHDF cells) were evaluated. Treatments with Chlorella and Spirulina lipid extracts in NHDF cells at 0.1, 0.5, and 1% significantly protect UVA-induced damage by decreasing oxidative stress in the form of reactive oxygen species (ROS) but do not display any significant protection from HEV-induced damage. Chlorella or Spirulina lipid extracts at 0.1 and 0.5% display significant anti-inflammatory effects by inhibiting the 2-propanol-induced IL-6 expression on NHDF cells in vitro but showed no decrease in TNF-α expression. Chlorella and Spirulina lipid extracts present antioxidant and anti-inflammatory activity on NHDF cells.

Separation of free fatty acids and neutral lipids from an aqueous suspension of crude microalgae oil with ethyl acetate

The aim of this study was to determine the liquid-liquid equilibrium data for a ternary system of water, ethyl acetate and oil from the microalgae Scenedesmus obliquus at 298.15, 308.15, and 318.15 K. The potential of the ternary system was also investigated for the selective extraction of free fatty acids and neutral lipids from crude microalgae oil. The equilibrium data were obtained using a model oil formulated from a mixture of refined vegetable oils, with a fatty acid profile analogous to the dark and opaque algae oil. The experimental equilibrium curves and tie lines were provided as a function of temperature for a free fatty acid content of 2%, 20% and 40% (w/w). The free fatty acids in the model oil migrated preferentially to the ethyl acetate-rich phase in each of the systems under study, with extraction efficiencies ranging from 79.4% to 97.1%. However, the extraction efficiencies for the neutral lipids were lower (51.5–70.1%), indicating the preferential extraction of free fatty acids. The distribution coefficients of neutral lipids (1.06–2.34) and free fatty acids (3.85–33.2) were determined in order to better understand the selectivity of the green solvent, ethyl acetate, in the purification of crude oil from algae. The selectivity values varied from 1.84 to 28.6, indicating that the solvent was effective in concentrating neutral lipids and free fatty acids in the ethyl acetate-rich phase. The increase in free fatty acid content therefore reduced the partition coefficient for neutral lipids, improving its selectivity, and making it possible to concentrate this lipid class by liquid-liquid separation with ethyl acetate.

Green Solvent-Based Extraction of Lipids and Proteins from Tenebrio molitor: Extraction Efficiency and Cytotoxic Activity

Green Solvent-Based Extraction of Lipids and Proteins from Tenebrio molitor: Extraction Efficiency and Cytotoxic Activity

Sequential extraction as a pioneering method to separate resin from bacuri fruit shell

Sequential extraction (SE) using supercritical CO2 plus ethanol was applied to separate resin from bacuri fruit shell, where process parameters such as particle size, pressure and cosolvent use were studied. The extracts obtained were also analyzed for phenolic compounds and antioxidant capacity. The extraction results showed that SE was able to separate the resin, which is the first report described in the literature, resulting in a patent. Furthermore, the smaller particle size (0.25 mm) showed the higher extraction rates, providing good yields of lipid extracts (up to 10.09 wt%) and ethanolic extracts (up to 13.78 wt%). The extracts presented good levels of phenolic compounds, which was associated with its high antioxidant capacity. Thus, the application of SE added value to bacuri fruit shell, enabling new strands for potential applications in the food, pharmaceutical and cosmetic industries, placing this by-product in the Amazon bioeconomy scenario.

Robust anti-tubercular profile of Solanum virginianum extract in enhancing isoniazid bioavailability and curtailing stress tolerance in Mycobacterium smegmatis.

The formidable survival mechanisms employed by Mycobacterium tuberculosis (Mtb), combined with the low bioavailability of anti-tubercular drugs and their associated hepatotoxicity, worsen tuberculosis management. Traditional medicinal plants offer potential solutions to these challenges. This study focuses on exploring the anti-tubercular potential of Solanum virginianum against Mycobacterium smegmatis, mc2155. HPTLC and UHPLC phytochemically characterized the hydro-methanolic extract of Solanum virginianum (SVE). SVE curtails the growth and viability of mc2155 under normal and in vitro stress conditions. The compromised cell wall integrity of mc2155 with SVE is depicted through scanning electron microscopy (SEM) while EtBr permeability assays and TLC-based comparative changes in lipids extraction addressed the integrity of the cell wall. Furthermore, SVE augmented the susceptibility of mc2155 towards Isoniazid (INH) through enhanced bioavailability. Adjunct treatment of SVE with INH demonstrated a markedly reduced survival of the intracellular bacilli. The study also uncovered the hepatoprotective potential of SVE in HepG2 cells. This research paves the way for deeper exploration into the potential of Solanum virginianum against virulent Mtb strains, emphasizing over the significance of traditional medicinal plants in tuberculosis treatment. Collectively, the findings suggest SVE as a potent candidate for independent or adjunct anti-tubercular therapy.

Automated green sample preparation for quantitative extraction of lipids in different sample matrices

An essential step in all analytical workflows is the quantitative and robust extraction of the molecules of interest. While typical lipid extractions use hazardous solvents from non-renewable sources, in this work, an extraction protocol utilizing green solvents such as ethanol and ethyl acetate is presented. The workflow was also automated to reduce variability and increase robustness as well as sample throughput. Moreover, this minimizes the contamination risks and lowers the operator's exposure to organic solvents. The developed workflow was critically evaluated by the recovery of various lipid standards from different matrices (human plasma, human serum, HepG2 cells) yielding quantitative values around 80–90 % for most lipid classes. The investigation into concentration dependency reveals the robustness of the method, demonstrating consistent recoveries across various lipid concentrations and matrix contents. In non-targeted lipidomics analyses, the green solvents proved to be comparable to established protocols that use conventional solvents such as methyl‑tert-butylether (MTBE) for lipid extraction.

Evaluation of the Pharmacological Efficiency of a Lipid Extract from the Tunic of the Marine Hydrobiont Halocynthia aurantium (Pallas, 1774)

Evaluation of the Pharmacological Efficiency of a Lipid Extract from the Tunic of the Marine Hydrobiont Halocynthia aurantium (Pallas, 1774)

Relationship between oligoarginine-induced membrane damage of single Escherichia coli cells and entry of the peptide into the cytoplasm

Cell-penetrating peptides (CPPs) can enter the cytosol of eukaryotic cells without killing them whereas some CPPs exhibit antimicrobial activity against bacterial cells. Here, to elucidate the mode of interaction of the CPP nona-arginine (R9) with bacterial cells, we investigated the interactions of lissamine rhodamine B red-labeled peptide (Rh-R9) with single Escherichia coli cells encapsulating calcein using confocal laser scanning microscopy. After Rh-R9 induced the leakage of a large amount of calcein, the fluorescence intensity of the cytosol due to Rh-R9 greatly increased, indicating that Rh-R9 induces cell membrane damage, thus allowing entry of a significant amount of Rh-R9 into the cytosol. To determine if the lipid bilayer region of the membrane is the main target of Rh-R9, we then investigated the interaction of Rh-R9 with single giant unilamellar vesicles (GUVs) comprising an E. coli polar lipid extract containing small GUVs and AlexaFluor 647 hydrazide (AF647) in the lumen. Rh-R9 entered the GUV lumen without inducing AF647 leakage, but leakage eventually did occur, indicating that GUV membrane damage was induced after the entry of Rh-R9 into the GUV lumen. The Rh-R9 peptide concentration dependence of the fraction of entry of Rh-R9 after a specific interaction time was similar to that of the fraction of leaking GUVs. These results indicate that Rh-R9 can damage the lipid bilayer region of a cell membrane, which may be related to its antimicrobial activity.