Wax Esters Research Articles

Effect of the extraction method on the lipid composition of purslane (Portulaca oleracea L.) seed oil

Purslane (Portulaca oleracea L.) is a widespread weed plant used since ancient times as remedy and as food. Its seed oil possesses significant bioactive potential due to the high content of important phytonutrients, mainly essential fatty acids and phytosterols. Although the total fatty acid composition of oil is well documented, till now there is no data published about individual lipid classes. The information on sterols is scarce as well. Therefore, the aim of this work was to characterize in details for the first time the individual lipid classes and their fatty acid composition, sterols, as well as the oxidative stability of the oil, depending on the extraction method by either hexane, chloroform–methanol mixtures or super-critical CO2. The results revealed no significant effect of extraction method on the individual lipid classes (monoacylglycerols, diacylglycerols, free fatty acids, free sterols, triacylglycerols, sterol esters, wax esters and hydrocarbons), including the fatty acid composition of saponifiables, and quite weak effect on the oxidative stability of the oil regarding super-critical CO2 vs. organic solvents. Detailed analyses of lipid composition confirmed the potential of purslane seed oil as a cheap and highly valuable source of phytonutrients as essential fatty acids and phytosterols, for application in food, pharmaceutical and cosmetic industries.

Post-transcriptional regulation of Dufour’s gland reproductive signals in bumble bees

Pheromone communication is a key mechanism by which the reproductive division of labor is maintained within insect communities. Understanding how pheromones evolved to regulate social behavior requires knowledge of the molecular regulation of their production. However, even in cases where pheromones were identified, our understanding of their biosynthesis and molecular regulation remains limited. Bumble bees provide a unique system to explore pheromone biosynthesis since workers produce ester sterility signals in their Dufour’s gland that differ from gyne-specific esters and are not produced by queens. These esters are hypothesized to be produced in the exocrine gland where they are stored, and indeed queens, gynes and workers differ significantly in the expression of Dufour’s gland genes coding to enzymes involved in the biosynthesis of esters. However, a previous transcriptome analysis revealed no gene expression differences in the Dufour’s gland of workers despite differences in both ester production and ovarian activation, suggesting that ester production may be regulated lower down. Proteomics of the Dufour’s gland of queens, gynes, and workers recovered over 2400 proteins and broadly matched the previous RNAseq data. However, more than 100 differentially expressed proteins were found between the worker groups, including key enzymes in fatty acid biosynthesis, indicating that the regulation of reproductive signal biosynthesis in workers is done post-transcription. Overall, our data provide evidence that pheromone biosynthesis in the Dufour’s gland is caste specific, that gynes and workers are likely using different enzymes to make their respective wax esters, and that the regulation on pheromone production in queens, gynes and workers is likely done at different regulatory levels, with workers signals being subjected to regulation at the protein level.

Integrative multi-omic analysis identifies genes associated with cuticular wax biogenesis in adult maize leaves.

Studying the genetic basis of leaf wax composition and its correlation with leaf cuticular conductance (gc) is crucial for improving crop productivity. The leaf cuticle, which comprises a cutin matrix and various waxes, functions as an extracellular hydrophobic layer, protecting against water loss upon stomatal closure. To address the limited understanding of genes associated with the natural variation of adult leaf cuticular waxes and their connection to gc, we conducted statistical genetic analyses using leaf transcriptomic, metabolomic, and physiological data sets collected from a maize (Zea mays L.) panel of ∼300 inbred lines. Through a random forest analysis with 60 cuticular wax traits, it was shown that high molecular weight wax esters play an important role in predicting gc. Integrating results from genome-wide and transcriptome-wide studies (GWAS and TWAS) via a Fisher's combined test revealed 231 candidate genes detected by all three association tests. Among these, 11 genes exhibit known or predicted roles in cuticle-related processes. Throughout the genome, multiple hotspots consisting of GWAS signals for several traits from one or more wax classes were discovered, identifying four additional plausible candidate genes and providing insights into the genetic basis of correlated wax traits. Establishing a partially shared genetic architecture, we identified 35 genes for both gc and at least one wax trait, with four considered plausible candidates. Our study enhances the understanding of how adult leaf cuticle wax composition relates to gc and implicates both known and novel candidate genes as potential targets for optimizing productivity in maize.

Dietary lipid is largely deposited in skin and rapidly affects insulating properties.

Skin has been shown to be a regulatory hub for energy expenditure and metabolism: mutations of skin lipid metabolism enzymes can change the rate of thermogenesis and susceptibility to diet-induced obesity. However, little is known about the physiological basis for this function. Here we show that the thermal properties of skin are highly reactive to diet: within three days, a high fat diet reduces heat transfer through skin. In contrast, a dietary manipulation that prevents obesity accelerates energy loss through skins. We found that skin was the largest target in a mouse body for dietary fat delivery, and that dietary triglyceride was assimilated both by epidermis and by dermal white adipose tissue. Skin from mice calorie-restricted for 3 weeks did not take up circulating lipids and showed a highly depleted stratum corneum. Dietary triglyceride acyl groups persist in skin for weeks after feeding. Using multi-modal lipid profiling, we have implicated both keratinocytes and sebocytes in the altered lipids which correlate with thermal function. In response to high fat feeding, wax diesters and ceramides accumulate, and triglycerides become more saturated. In contrast, in response to the dramatic loss of adipose tissue that accompanies restriction of the branched chain amino acid isoleucine, skin becomes more heat-permeable, resisting changes induced by Western diet feeding, with a signature of depleted signaling lipids. We propose that skin should be routinely included in physiological studies of lipid metabolism, given the size of the skin lipid reservoir and its adaptable functionality.

Meibomian gland lipid alterations and ocular surface sequela in Awat2 knockout murine model of meibomian gland dysfunction and evaporative dry eye disease

PurposeThere is an urgent need for animal models of meibomian gland dysfunction (MGD) and evaporative dry eye disease (EDED) to understand their pathophysiology and investigate novel therapeutics. This study sought to further define the acyl-CoA: wax alcohol acyltransferase 2 knockout (Awat2 KO) mouse as a model of EDED using a combination of novel clinical, biochemical, and biophysical endpoints. MethodsWildtype and Awat2 KO mice between 1 and 18 months of age were used. Ocular examinations and advanced imaging were performed. The lipidomic composition and in situ melting temperature of meibum were determined. qPCR was performed to define ocular surface gene and pro-inflammatory transcript expression. Dynamic contact angle goniometry was performed to assess the adherence capability of the ocular surface. ResultsAwat2 KO mice have mild, white, hyperreflective corneal opacities of the anterior stroma and significantly enlarged apical epithelial cells (P=0.0004). In Awat2 KO meibum, wax esters were 9-10 times lower than in wildtype meibum. Additionally, meibum melting temperature increased from 32° to 47°C (P<0.0001), leading to impaired meibum secretion and dilation of the central duct. Awat2 KO corneal epithelia had significantly decreased mucin expression (Muc1 and Muc4, P=0.0043) and increased interferon-γ production (P=0.0303). Awat2 KO globes have a significantly shortened time of droplet adherence to their ocular surface (P=0.0053), indicating a decreased tear film adherence capacity. Wildtype corneal epithelia does not express Awat2, indicating that the EDED phenotype is secondary to the loss of Awat2 from the meibomian glands. ConclusionsAwat2 KO mice recapitulate many of features of human MGD and EDED, representing a model to test novel therapeutics.

The dominant white color trait of the melon fruit rind is associated with epicuticular wax accumulation.

Microscopic analyses and chemical profiling demonstrate that the white rind phenotype in melon fruit is associated with the accumulation of n-alkanes, fatty alcohols, aldehydes and wax esters. Serving as an indicator of quality, the rind (or external) color of fruit directly affects consumer choice. A fruit's color is influenced by factors such as the levels of pigments and deposited epicuticular waxes. The latter produces a white-grayish coating often referred to as "wax bloom". Previous reports have suggested that some melon (Cucumis melo L.) accessions may produce wax blooms, where a dominant white rind color trait was genetically mapped to a major locus on chromosome 7 and suggested to be inherited as a single gene named Wi. We here provide the first direct evidence of the contribution of epicuticular waxes to the dominant white rind trait in melon fruit. Our light and electron microscopy and gas chromatography-mass spectrometry (GC-MS) comparative analysis of melon accessions with white or green rinds reveals that the rind of melon fruit is rich in epicuticular waxes. These waxes are composed of various biochemical classes, including fatty acids, fatty alcohols, aldehydes, fatty amides, n-alkanes, tocopherols, triterpenoids, and wax esters. We show that the dominant white rind phenotype in melon fruit is associated with increased accumulation of n-alkanes, fatty alcohols, aldehydes and wax esters, which are linked with the deposition of crystal-like wax platelets on their surfaces. Together, this study broadens the understanding of natural variation in an important quality trait of melon fruit and promotes the future identification of the causative gene for the dominant white rind trait.

Lipid composition of coral propagules and reproductive material in coral restoration nurseries

Coral restoration efforts have rapidly increased worldwide, including the development of several programmes on the Great Barrier Reef (GBR) in recent years. While many restoration programmes utilise in-water nurseries to accelerate coral biomass yields, the impact of nursery environments on propagule quality has not been examined despite the importance of coral fitness for ensuring resistant populations. Here, we investigated two fitness indicators (lipid diversity and tissue protein abundance) of Acropora millepora adults and eggs grown on coral nurseries versus native reef on the GBR, with adults assessed at two sites (Blue Lagoon and Rayban) and eggs assessed at one site (Blue Lagoon). Lipid profiles of adult colonies varied by site and origin (nursery versus wild reef), with adult nursery corals exhibiting an elevated relative abundance of storage lipids (diacylglycerols and triacylglycerols) and lipid classes responsible for regulating membrane structure (phosphatidylcholines and sterol esters), while wild corals were characterised by a greater relative abundance of fatty acids and classes involved in immunoregulation. Comparing eggs from different origins, nursery offspring were richer in energy-storing triacylglycerols, as well as ceramides and phosphatidylcholines essential for membrane structure, while wild eggs had a greater relative abundance of wax ester species also important for energy storage. No differences were found in total protein abundance (adult or eggs) or egg physical characteristics (count and size) between nursery and wild origins. Variations in lipid profiles are consistent with differences in environmental conditions between reef sites and origin (nursery versus wild), highlighting the need to consider site selection and propagation conditions when planning restoration projects. Importantly, these findings demonstrate that the lipid classes with the highest relative abundance in A. millepora nursery and wild eggs differed from those in adults from the same origin, suggesting that propagation origin is more important for driving lipid profiles in coral eggs compared to parental effects.

Influence of Lipid Class Used for Omega-3 Fatty Acid Supplementation on Liver Fat Accumulation in MASLD.

Metabolic dysfunction-associated steatotic liver disease (MASLD) occurs in subjects with obesity and metabolic syndrome. MASLD may progress from simple steatosis (i.e., hepatic steatosis) to steatohepatitis, characterized by inflammatory changes and liver cell damage, substantially increasing mortality. Lifestyle measures associated with weight loss and/or appropriate diet help reduce liver fat accumulation, thereby potentially limiting progression to steatohepatitis. As for diet, both total energy and macronutrient composition significantly influence the liver's fat content. For example, the type of dietary fatty acids can affect the metabolism of lipids and hence their tissue accumulation, with saturated fatty acids having a greater ability to promote fat storage in the liver than polyunsaturated ones. In particular, polyunsaturated fatty acids of n-3 series (omega-3), such as docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), have been intensively studied for their antisteatotic effects, both in preclinical animal models of obesity and hepatic steatosis and in overweight/obese patients. Their effects may depend not only on the dose and duration of administration of omega-3, or DHA/EPA ratio, but also on the lipid class used for their supplementation. This review summarizes the available evidence from recent comparative studies using omega-3 supplementation via different lipid classes. Albeit the evidence is mainly limited to preclinical studies, it suggests that phospholipids and possibly wax esters could provide greater efficacy against MASLD compared to traditional chemical forms of omega-3 supplementation (i.e., triacylglycerols, ethyl esters). This cannot be attributed solely to improved EPA and/or DHA bioavailability, but other mechanisms may be involved. Keywords: MASLD • Metabolic dysfunction-associated steatotic liver disease • NAFLD • Non-alcoholic fatty liver disease • n-3 polyunsaturated fatty acids.

Systematic literature review of acne vulgaris, its sophisticated treatment approaches, types, treatment and their side effects.

Background: Acne formation can be controlled effectively but it heals slowly and has numerous interactions with neuropeptides. Once gone another one pops up. Sometimes it can be filled with pus. Its size varies and can cause anxiety, depression, stress and social withdrawal. The mechanism is hyper secretion of sebaceous glands. Introduction: Acne vulgaris is a condition when hair follicles are accumulated with oil, dead cells and bacteria from skin. It can lead to formation of blackhead, whitehead, oily skin, pimples and scarring eventually. Patients have raised sebum level and thus skin is more prone to acne formation; triggered by propionibacterium, genetics, environment and food. Most commonly occurring on the face, upper limb and trunk between teen to middle age. Sebum contains wax esters, cholesterol, and triglyceride. Objectives: · To extensively explain the etiology of acne vulgaris, hormonal, bacterial causes and UV light. · To write comprehensive review on treatment of AV conventional, topical, retinoid and antibiotics based treatment. · Write a comprehensive review on its complications and its management. Methodology: In this research article we have explained systematic review, as well as we have described the systematic literature review of Acne Vulgaris, We did a literature search using electronic bibliographic databases and Journals : such as ELSEVIER ,British Medical Journals ,MEDLINE (Ovid SP and PubMed), EMBASE, The Cochrane Library (Cochrane Database of Systematic Reviews (CDSR), and Cochrane Central Register of Controlled Trials (CENTRAL), as well as annual meetings abstracts from inception till Oct 2024. The literature search has focused mainly on randomized clinical trials, Meta-analysis, phase II/III, and retrospective studies. Results and Conclusion: The review included 45 studies with a total of 12000 participants. Acne vulgaris affects 9% of the population worldwide, 85% are between 12-24 years. Characters such as age and sex may vary among different studies. Many papers demonstrate the effect of demographic factors. Reaching late teenage can decrease its prevalence, and eruption is higher in females compared to male. Topical retinoid and benzoyl per oxide were effective in reducing inflammatory and non-inflammatory lesions. For mild to moderate acne pharmacological treatment, chemical, antibiotic and photodynamic therapy are among most effective. Oral antibiotics while effective, have raised concerned about resistance. For moderate to severe, the most effective one is oral Isotretinoin and photodynamic treatment. Most of us already know too much about this topic. Its 8th commonest disease known globally. It has many treatments. It’s the cause of social withdrawal and depression. In this case it is important to know about the causes that shift non inflammatory to inflammatory ones. Dermatologists are trying their best in combination of multiple therapies to treat it successfully. Personalized treatment plans considering patient-specific factors, such as acne severity, patient preference and risk of adverse effects, are essential for optimal management.

Plant-Based Oils for Sustainable Lubrication Solutions—Review

Traditional lubricants, often containing harmful chemicals and synthetic or fossil-derived oils, pose environmental risks by damaging ecosystems and threatening human health and wildlife. There is a growing demand for environmentally sustainable and cost-effective bio-based lubricants derived from renewable raw materials. These bio-based oils often possess natural lubricating properties, making them an attractive alternative to traditional synthetic lubricants. In addition to providing effective lubrication, they offer good biodegradability and minimal toxicity, which are essential for reducing environmental impact. However, the primary challenge lies in optimizing their performance to match or surpass that of conventional lubricants while ensuring they remain cost-effective and widely available. This paper reviews the general requirements for lubricants and explores how plant-based oils can be utilized to meet the diverse lubrication needs across various industries. Further, it highlights different approaches that can be used for further improvements in the area of plant-based lubrication through bio-inspired means, such as the use of estolides, wax esters, or erucic acid, as well as through additions of nanomaterials, such as nanoparticles, nanoclays, or two-dimensional films.

Recent Advances in Food Waste Transformations into Essential Bioplastic Materials.

Lignocellulose is a major biopolymer in plant biomass with a complex structure and composition. It consists of a significant amount of high molecular aromatic compounds, particularly vanillin, syringeal, ferulic acid, and muconic acid, that could be converted into intracellular metabolites such as polyhydroxyalkanoates (PHA) and hydroxybutyrate (PHB), a key component of bioplastic production. Several pre-treatment methods were utilized to release monosaccharides, which are the precursors of the relevant pathway. The consolidated bioprocessing of lignocellulose-capable microbes for biomass depolymerization was discussed in this study. Carbon can be stored in a variety of forms, including PHAs, PHBs, wax esters, and triacylglycerides. From a biotechnology standpoint, these compounds are quite adaptable due to their precursors' utilization of hydrogen energy. This study lays the groundwork for the idea of lignocellulose valorization into value-added products through several significant dominant pathways.

Genome editing-based mutagenesis stably modifies composition of wax esters synthesized by Euglena gracilis under anaerobic conditions

Microalgal oil production represents a promising renewable biofuel source. Metabolic engineering can enhance its utility, transforming it into an improved biofuel and expanding its applications as a feedstock for commodity chemicals, thereby increasing their value in biorefineries. This study focused on anaerobic wax ester production by the microalga Euglena gracilis, aiming to develop stable mutant strains with altered wax ester profiles through genome editing. Two enzymes in the fatty acid beta-oxidation pathway involved in wax ester production were targeted—3-ketoacyl-CoA thiolase and acyl-CoA dehydrogenase—using clustered regularly interspaced short palindromic repeats/Cas9. The results revealed one genetic mutation that lengthened and three that shortened the distribution of wax ester compositions compared to the wild-type (WT). The triple-knockout mutant, combining mutations that shorten wax ester chains, produced wax esters with acyl chains two carbons shorter than WT. This study established a methodology to stably modify wax ester composition in E. gracilis.

Safety assessment of the substances 'wax, rice bran, oxidised' and 'wax, rice bran, oxidised, calcium salt' for use in food contact materials.

The EFSA Panel on Food Contact Materials (FCM) assessed the safety of the substances 'wax, rice bran, oxidised' and 'wax, rice bran, oxidised, calcium salt', used as additives up to 0.3% in polyethylene terephthalate (PET), polyamide (PA), thermoplastic polyurethane (TPU), polylactic acid (PLA) and poly(vinyl chloride) (PVC) in contact with all food types for long-term storage at room temperature and below, after hot-fill and/or heating. The substances consist of the chemical classes wax esters, carboxylic acids, alcohols and calcium salts of acids, along with an unidentified organic fraction up to ■■■■■ w/w. Migration into 10% ethanol and 4% acetic acid was below 0.012 mg/kg for each chemical class, and about 0.001 mg/kg for the unidentified fraction. In isooctane, migration was up to 0.297 mg/kg food for wax esters, below 0.01 mg/kg food for the other chemical classes and about 0.02 mg/kg food for the unidentified fraction. The contact with dry food and food simulated by 20% ethanol were considered covered by the migration tests with aqueous simulants. Based on genotoxicity assays and compositional analyses, the constituents of the chemical classes did not raise a concern for genotoxicity. The potential migration of individual constituents or groups of chemically-related compounds of the unidentified fraction would result in exposures below (for aqueous food) and above (for fatty food) the threshold of toxicological concern for genotoxic carcinogens. Therefore, the FCM Panel concluded that the substances are not of safety concern for the consumer, if used as additives up to 0.3% w/w in PET, PLA and rigid PVC materials and articles intended for contact with all food types except for fatty foods, for long-term storage at room temperature and below, including hot-fill and/or heating up to 100°C for up to 2 h.

Using kraft lignin nanoparticles for the stabilization of nano/micro wax carriers

Due to its amphiphilic structure, lignin has the potential to stabilize emulsions via adsorption at the oil/water interface. By converting lignin into nanoparticles, we can significantly enhance its emulsion-stabilizing capabilities through a Pickering-type stabilization mechanism. Two essential elements may be modified to fine-tune emulsion stability: the size of the lignin nanoparticles (LNPs) and the physicochemical nature of the lipid phase. In this context, we highlight the behavior and utility of unmodified LNPs in the preparation of Pickering emulsions made up of water and a complex bio-based pharmaceutical-grade wax that can be used for the formulation of lipid carriers. As a proof-of-concept, we employ the developed Pickering emulsions to encapsulate indocyanine green (ICG), an FDA-approved dye commonly used in medical imaging applications. We demonstrate that ultra-small LNPs are well-suited for the colloidal stabilization of pharmaceutical wax ester micro beads. This stabilization does not require any lignin modification. Additionally, we present evidence that our new lipid/lignin hybrid carrier has potential as a new drug delivery system.

MALDI imaging unveils spatial lipidomics dynamics during jojoba seed germination and post-germination

Jojoba (Simmondsia chinensis Schneider), the sole species in the Simmondsiaceae family, holds significant economic value. Understanding the mechanism of seed germination and post-germination is essential for jojoba breeding programs. For the first time, this study employed matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) to comprehensively detect and image endogenous lipids in jojoba seeds during germination and post-germination. Meanwhile, a new method of lithium salt-doped 2-mercaptobenzothiazole (2-MBT) matrix was adopted to detect the wax ester in jojoba seeds. The spatial lipidomic analysis of MALDI-MS revealed the dynamics of lipid alterations during three stages of jojoba seed (stage I, quiescent seed; stage II, germination; stage III, post-germination). MALDI-MSI results demonstrated heterogeneous distributions of differential lipids and main wax esters (WEs) in unique tissue regions of the jojoba seed at different stages. For instance, at stages II and III, three phosphatidylcholines (PCs) (PC(36:2), PC(36:4), and PC(35:3)) are mainly localized in the cotyledons, while the other two PCs (PC(34:1) and PC(36:1)) were distributed in both cotyledons and embryonic axis. In contrast, three phosphatidylethanolamines (PE(36:1), PE(42:4), and PE(42:9)) are primarily distributed near the embryonic axis and phosphatidylserine (PS(32:0)) was found exclusively in the embryonic axis area at three stages. In addition, three phosphatidic acids (PA(40:7), PA(46:2), and PA(42:1)) displayed highly accumulated in the middle of the cotyledons at stages II and III. Besides, WEs are continuously degraded mainly in the cotyledons during germination and post-germination processes. This study offers new insights into lipid alterations from a spatial lipidomics perspective and can contribute to our understanding of the role of these specific lipids during jojoba seed germination and post-germination. These findings will deepen our understanding of the metabolic processes involved in jojoba seed germination and seedling development and have underlying implications for crop improvement and seed quality control.

Usefulness of the 1H NMR Multisuppression Approach for the Global Characterization of Monovarietal Extra-Virgin Olive Oils.

Extra-virgin olive oil (EVOO) is one of the most appreciated vegetable oils worldwide, but its high price makes it prone to suffer adulteration with lower quality oils. Therefore, it is important to have methodologies able to study EVOO composition as a whole in a simple and fast way, in order to guarantee its quality and safety. For this purpose, in this study, commercial samples of five Spanish olive cultivars (Arbequina, Arroniz, Cornicabra, Hojiblanca, Picual) were studied by Proton Nuclear Magnetic Resonance (1H NMR) spectroscopy, using standard and multisuppression pulses. The aim was to explore the possibility of 1H NMR use to characterize in a single run and in a global way the composition of these monocultivar oils, regarding not only their main components (fatty acids supported on triglycerides) but also minor ones (squalene, sterols, diterpenic wax esters of phytol and geranylgeraniol, phenolic and secoiridoid derivatives, like tyrosol, hydroxytyrosol, oleacein, oleocanthal, and lignans, among others, and aldehydes). The use of univariate and multivariate statistical analyses confirmed the presence of compositional features that were specific to some olive varieties. The Arbequina and Arroniz oils showed the most characteristic features that allowed for clearly differentiating them from the others. In contrast, the discrimination between the Cornicabra, Hojiblanca and Picual oils was not so easily achieved.

Structural characterization of wax esters using ultraviolet photodissociation mass spectrometry

Wax esters play critical roles in biological systems, serving functions from energy storage to chemical signaling. Their diversity is attributed to variations in alcohol and acyl chains, including their length, branching, and the stereochemistry of double bonds. Traditional analysis by mass spectrometry with collisional activations (CID, HCD) offers insights into acyl chain lengths and unsaturation level. Still, it falls short in pinpointing more nuanced structural features like the position of double bonds. As a solution, this study explores the application of 213-nm ultraviolet photodissociation (UVPD) for the detailed structural analysis of wax esters. It is shown that lithium adducts provide unique fragments as a result of Norrish and Norrish-Yang reactions at the ester moieties and photoinduced cleavages of double bonds. The product ions are useful for determining chain lengths and localizing double bonds. UVPD spectra of various wax esters are presented systematically, and the effect of activation time is discussed. The applicability of tandem mass spectrometry with UVPD is demonstrated for wax esters from natural sources. The UHPLC analysis of jojoba oil proves the compatibility of MS2 UVPD with the chromatography time scale, and a direct infusion is used to analyze wax esters from vernix caseosa. Data shows the potential of UVPD and its combination with CID or HCD in advancing our understanding of wax ester structures.Graphical

Two protocols for the detection of oleaginous bacteria using Oil Red O

The selection of oleaginous bacteria, potentially applicable to biotechnological approaches, is usually carried out by different expensive and time-consuming techniques. In this study, we used Oil Red O (ORO) as an useful dye for staining of neutral lipids (triacylglycerols and wax esters) on thin-layer chromatography plates. ORO could detect minimal quantities of both compounds (detection limit, 0.0025 mg of tripalmitin or 0.005 mg of cetylpalmitate). In addition, we developed a specific, rapid, and inexpensive screening methodology to detect triacylglycerol-accumulating microorganisms grown on the agar plate. This staining methodology detected 9/13 strains with a triacylglycerol content higher than 20% by cellular dry weight. ORO did not stain polyhydroxyalkanoates-producing bacteria. The four oleaginous strains not detected by this screening methodology exhibited a mucoid morphology of their colonies. Apparently, an extracellular polymeric substance produced by these strains hampered the entry of the lipophilic dye into cells. The utilization of the developed screening methodology would allow selecting of oleaginous bacteria in a simpler and faster way than techniques usually used nowadays, based on unspecific staining protocols and spectrophotometric or chromatographic methods. Furthermore, the use of ORO as a staining reagent would easily characterize the neutral lipids accumulated by microorganisms as reserve compounds.Key points• Oil Red O staining is specific for triacylglycerols• Oil Red O staining is useful to detect oleaginous bacteria• Fast and inexpensive staining to isolate oleaginous bacteria from the environment

Lipid analyses of oil-bearing biomass using a thermally induced derivatization method

This study assessed the efficacy of a commercial fatty acid (FA) methylation kit for analyzing insect/microalgal biomass lipids by comparing it to a thermally induced derivatization method, which is known for its high conversion yield and impurity resistance and serves as a benchmark for evaluating kit performance, as a comparative standard. In insect lipid analysis, kit-based derivatization demonstrated a lower yield of FA methyl esters (FAMEs) than that of the thermally induced method, which is attributable to inherent technical limitations (vulnerability to impurities). This yield discrepancy was further highlighted in microalgal lipid analyses. The efficiency of FA derivatization using the kit-based method was contingent on the lipid structure, as evidenced by the diminished conversion yields for wax esters in Euglena gracilis. Furthermore, for Chlamydomonas reinhardtii, the lower FAME yield observed with kit-based derivatization suggested potential inadequacies in lipid extraction, particularly in the presence of rigid cellular structures such as cell walls. This reduced interaction between lipids and methanol could lead to diminished FAME conversion yields. These findings highlight the notable influence of the biomass type and lipid structure on the efficiency of kit-based derivatization, potentially yielding suboptimal results in lipid analysis.

Enhancing Erucic Acid and Wax Ester Production in Brassica carinata through Metabolic Engineering for Industrial Applications

Metabolic engineering enables oilseed crops to be more competitive by having more attractive properties for oleochemical industrial applications. The aim of this study was to increase the erucic acid level and to produce wax ester (WE) in seed oil by genetic transformation to enhance the industrial applications of B. carinata. Six transgenic lines for high erucic acid and fifteen transgenic lines for wax esters were obtained. The integration of the target genes for high erucic acid (BnFAE1 and LdPLAAT) and for WEs (ScWS and ScFAR) in the genome of B. carinata cv. ‘Derash’ was confirmed by PCR analysis. The qRT-PCR results showed overexpression of BnFAE1 and LdPLAAT and downregulation of RNAi-BcFAD2 in the seeds of the transgenic lines. The fatty acid profile and WE content and profile in the seed oil of the transgenic lines and wild type grown in biotron were analyzed using gas chromatography and nanoelectrospray coupled with tandem mass spectrometry. A significant increase in erucic acid was observed in some transgenic lines ranging from 19% to 29% in relation to the wild type, with a level of erucic acid reaching up to 52.7%. Likewise, the transgenic lines harboring ScFAR and ScWS genes produced up to 25% WE content, and the most abundant WE species were 22:1/20:1 and 22:1/22:1. This study demonstrated that metabolic engineering is an effective biotechnological approach for developing B. carinata into an industrial crop.