Lipid Modification Research Articles

Cold plasma bubbling: Impact on safety, physicochemical properties, and nutritional quality of sesame milk

The decline in the consumption of animal milk raises the possibility of using different plant materials in producing vegan milk. Sesame is an oilseed, rich in nutrients, making it a suitable animal milk substitute. However, microbial stability, quality, and anti-nutritional factors must be considered while processing vegan milk. In this study, plasma bubbling (PB) was used for processing sesame milk (at various exposure times-10, 20 and 30 min). PB effectively reduced the microbial load (1.82 and 0.64 log reduction of coliforms and bacterial load), yet it increased oxidation of lipids and reduction in whiteness index. However, the reduction in phytates (1.48–0.58 mg/g), oxalates (39.13–19.96 μg/g) and lipoxygenase activity (18.3 from 100%) makes it an efficient alternative technique in the processing of sesame milk. Further, studying the structural modifications of protein and lipids would be beneficial in understanding plasma influence in the processing of sesame milk.

Protein interactomes for plant lipid biosynthesis and their biotechnological applications.

Plant lipids have essential biological roles in plant development and stress responses through their functions in cell membrane formation, energy storage and signalling. Vegetable oil, which is composed mainly of the storage lipid triacylglycerol, also has important applications in food, biofuel and oleochemical industries. Lipid biosynthesis occurs in multiple subcellular compartments and involves the coordinated action of various pathways. Although biochemical and molecular biology research over the last few decades has identified many proteins associated with lipid metabolism, our current understanding of the dynamic protein interactomes involved in lipid biosynthesis, modification and channelling is limited. This review examines advances in the identification and characterization of protein interactomes involved in plant lipid biosynthesis, with a focus on protein complexes consisting of different subunits for sequential reactions such as those in fatty acid biosynthesis and modification, as well as transient or dynamic interactomes formed from enzymes in cooperative pathways such as assemblies of membrane-bound enzymes for triacylglycerol biosynthesis. We also showcase a selection of representative protein interactome structures predicted using AlphaFold2, and discuss current and prospective strategies involving the use of interactome knowledge in plant lipid biotechnology. Finally, unresolved questions in this research area and possible approaches to address them are also discussed.

RNA sequencing analysis of the longissimus dorsi to identify candidate genes underlying the intramuscular fat content in Anqing Six-end-white pigs.

Intramuscular fat (IMF) is a significant marker for pork quality. The Anqing Six-end-white pig has the characteristics of high meat quality and IMF content. Owing to the influence of European commercial pigs and a late start in resource conservation, the IMF content within local populations varies between individuals. This study analyzed the longissimus dorsi transcriptome of purebred Anqing Six-end-white pigs with varying IMF content to recognize differentially expressed genes. We identified 1528 differentially expressed genes between the pigs with high (H) and low (L) IMF content. Based on these data, 1775 Gene Ontology terms were significantly enriched, including lipid metabolism, modification and storage, and regulation of lipid biosynthesis. Pathway analysis revealed 79 significantly enriched pathways, including the Peroxisome proliferator-activated receptor and mitogen-activated protein kinase signaling pathways. Moreover, gene set enrichment analysis indicated that the L group had increased the expression of genes related to ribosome function. Additionally, the protein-protein interaction network analyses revealed that VEGFA, KDR, LEP, IRS1, IGF1R, FLT1 and FLT4 were promising candidate genes associated with the IMF content. Our study identified the candidate genes and pathways involved in IMF deposition and lipid metabolism and provides data for developing local pig germplasm resources.

Copper and Nickel Induce Changes in the Lipid and Fatty Acid Composition of Anodonta cygnea.

The effect of copper and nickel ions on the lipid composition of freshwater mussels Anodonta cygnea was investigated using an aquarium experiment. The contents of the main lipid classes were determined using thin layer chromatography and spectrophotometry, and the fatty acid composition was analysed using gas-liquid chromatography. The results indicated that copper and nickel had different effects on the mussels' lipid composition, with copper producing less effect on the composition of lipids and fatty acids than nickel. On the first experiment day, excessive copper content in the organism caused oxidative stress and modifications in membrane lipids, which returned to their initial level by the end of the experiment. Nickel accumulated predominantly in gills; however, significant modifications in lipids and fatty acids were seen also in the digestive gland from the first day of the experiment. This indicated the activation of nickel-induced lipid peroxidation processes. Moreover, this study revealed a dose-dependent effect of nickel on lipid composition, which was likely related to the development of compensatory biochemical mechanisms in response to nickel-induced oxidative stress. A comparative study of the lipid composition alteration in mussels in response to copper and nickel action revealed the consequences of the toxic impact of metal ions and the defensive mechanisms that organisms employ to detoxify and remove xenobiotics.

Engineered liposomes to deliver nucleic acid mimics in Escherichia coli.

Antisense oligonucleotides (ASOs) composed of nucleic acid mimics (NAMs) monomers are considered as potential novel therapeutic drugs against bacterial infections. However, bacterial envelopes are generally impermeable to naked oligonucleotides. Herein, liposomes loaded with NAMs-modified oligonucleotides (LipoNAMs) were evaluated to deliver ASOs in Escherichia coli. Specifically, we tested several surface modifications that included methoxyPEG conjugated to different lipid anchors or modification of the PEG distal ends with maleimide groups and antibodies. MethoxyPEG coated LipoNAMs showed low delivery efficiency for most bacteria, but maleimide-functionalized PEG LipoNAMs were able to deliver ASOs to nearly half of the bacterial population. Conjugation of antibodies to maleimide-functionalized PEG LipoNAMs increased 1.3-fold the delivery efficiency, enhancing the selectivity towards E. coli and biocompatibility. This work demonstrated for the first time that the coupling of antibodies to PEGylated liposomes can significantly improve the delivery of ASOs in E. coli, which might bring alternative routes for the treatment of bacterial infections in the future.

US public investment in development of mRNA covid-19 vaccines: retrospective cohort study

ObjectiveTo estimate US public investment in the development of mRNA covid-19 vaccines.DesignRetrospective cohort study.SettingPublicly funded science from January 1985 to March 2022.Data sourcesNational Institutes of Health (NIH) Report Portfolio Online...

Post-translational palmitoylation of metabolic proteins.

Numerous cellular proteins are post-translationally modified by addition of a lipid group to their structure, which dynamically influences the proteome by increasing hydrophobicity of proteins often impacting protein conformation, localization, stability, and binding affinity. These lipid modifications include myristoylation and palmitoylation. Palmitoylation involves a 16-carbon saturated fatty acyl chain being covalently linked to a cysteine thiol through a thioester bond. Palmitoylation is unique within this group of modifications, as the addition of the palmitoyl group is reversible and enzyme driven, rapidly affecting protein targeting, stability and subcellular trafficking. The palmitoylation reaction is catalyzed by a large family of Asp-His-His-Cys (DHHCs) motif-containing palmitoyl acyltransferases, while the reverse reaction is catalyzed by acyl-protein thioesterases (APTs), that remove the acyl chain. Palmitoyl-CoA serves an important dual purpose as it is not only a key metabolite fueling energy metabolism, but is also a substrate for this PTM. In this review, we discuss protein palmitoylation in regulating substrate metabolism, focusing on membrane transport proteins and kinases that participate in substrate uptake into the cell. We then explore the palmitoylation of mitochondrial proteins and the palmitoylation regulatory enzymes, a less explored field for potential lipid metabolic regulation.

A multicenter, prospective, randomized controlled trial of intracranial hemorrhage risk of intensive statin therapy in patients with acute ischemic stroke combined with cerebral microbleeds (CHRISTMAS): Study protocol.

Low serum levels of major lipid markers have been proved to be significantly associated with increased risks of hemorrhagic stroke (HS) and cerebral microbleeds (CMBs). However, there is no lipid modification guideline telling us how to maintain a balance between the prevention of ischemic stroke recurrence and the prevention of hemorrhagic events, especially in patients with acute ischemic stroke (AIS) and CMBs. The Intracranial Hemorrhage Risk of Intensive Statin Therapy in Patients with Acute Ischemic Stroke combined with Cerebral Microbleeds (CHRISTMAS) trial evaluates the risk of intracranial hemorrhage (i.e., HS and CMBs) of high-dose statin therapy in patients with AIS combined with CMBs. This is an investigator-initiated, multicenter, prospective, randomized controlled clinical trial design. Up to 344 eligible patients will be consecutively randomized to receive high-dose or low-dose atorvastatin in 1:1 ratio in 5 stroke centers in China. CHRISTMAS trial has co-primary outcomes, namely, hemorrhage risk: the incidence of HS and the changes in degree of CMBs until the end of 36-month follow-up. The primary hypothesis of this study is that an excessive reduction in serum lipid levels by an intensive statin therapy in AIS patients with CMBs can increase the risk of intracranial hemorrhage. This study will shed light on new clinical decisions regarding the long-term serum lipid management in these patients with dilemma in clinical practice. Clinicaltrials.gov, identifier: NCT05589454.

The structural basis of lipid A modification with aminoarabinose revealed by cryoEM structures of S. enterica ArnT.

Lipid A is the major lipidic component of the bacterial lipopolysaccharide (LPS) decorating the outer membrane of Gram-negative (GN) bacteria. Modification of Lipid A by addition of diverse chemical moieties leads to altered host recognition, evasion of host defenses, and resistance to antimicrobial agents. In particular, modification of the phosphates of Lipid A with the aminoarabinose moiety 4-amino-4-deoxy-L-arabinose (L-Ara4N) leads to charge modification of the outer bacterial membrane, resistance against natural cationic antimicrobial peptides (CAMPs), and is the major contributor for development of polymyxin resistance in Escherichia coli and Salmonella enterica. Polymyxins are currently used as last resort antibiotics against multidrug resistant (MDR) GN bacteria, either as monotherapies or in combination with other antibiotics. The aminoarabinose transferase ArnT, the final enzyme in the aminoarabinose biosynthetic pathway responsible for the biosynthesis of L-Ara4N, catalyzes the transfer of the aminoarabinose sugar from a donor lipid to the phosphates of Lipid A. Using single particle cryo-electron microscopy (cryoEM) we have determined the structure of ArnT from Salmonella enterica in three states: i) bound to both the acceptor Lipid A and the donor undecaprenyl phosphate (UndP), ii) bound to the acceptor Lipid A alone, after mutating one of the residues coordinating UndP, and iii) bound to both the acceptor Lipid A and the full sugar-bound donor, by expressing a catalytically-dead mutant of ArnT. These structures are the first to capture the Lipid A-bound state(s) of ArnT. They allow us to fully characterize substrate binding, and to accurately localize the active site of the enzyme. By comparing these structures to existing structures of ArnT from C. metallidurans, we provide further insights towards understanding the structural basis of catalysis and the substrate binding cycle of the aminoarabinose transferase ArnT.

FacePalm: Probing dynamic S-palmitoylation of CaV channel complexes using engineered depalmitoylases.

S-palmitoylation is a powerful post-translational modification of ion channels that tunes its function and localization. Unlike other lipid modifications, palmitoylation is a reversible process orchestrated by zDHHC family of protein palmitoyl-transferases and acyl-protein thioesterases that attach and excise palmitic groups, respectively. Even so, dissecting spatial and temporal dynamics of palmitoylation of large channel complexes remains challenging, owing to limitations in molecular approaches to acutely manipulate palmitoylation of specific targets. Here, we develop a new approach to rapidly and inducible depalmitoylate target proteins by engineering the ABHD17 (α/β-hydrolase domain containing protein 17) family of protein depalmitoylases with an FKBP/FRBP chemically inducible dimerization system. This approach is termed FKBP/FRB-activated chemogenetic excision of Palmitoylation or FacePalm. Application of FacePalm revealed multifaced regulation of CaV1/2 channel complexes by palmitoylation. First, time-lapse confocal imaging revealed dynamic redistribution of the CaVβ2A subunit from the plasma membrane to the cytoplasm. Second, whole-cell and single-channel electrophysiology show differences in both inactivation and activation of CaV2 channels depending on palmitoylation of distinct sites on either the β2A or the pore-forming α-subunit. Third, targeting FacePalm to distinct subcellular compartments revealed intrinsic differences in the rate of depalmitoylation of CaVβ2A subunit in different organelles. To further demonstrate generality, we applied FacePalm to various cellular proteins and found substrate-specific differences in palmitoylation dynamics. In all, these studies provide critical insights into the exquisite molecular and cellular mechanisms regulating CaV channel function. Furthermore, FacePalm opens new frontiers to dissect dynamic palmitoylation of ion channel complexes in distinct physiological settings.

Oxidative Stress Contributes to Inflammatory and Cellular Damage in Systemic Lupus Erythematosus: Cellular Markers and Molecular Mechanism.

Systemic lupus erythematosus (SLE) is a multifactorial autoimmune disease with complex pathogenesis, the treatment of which relies exclusively on the use of immunosuppressants. Increased oxidative stress is involved in causing inflammatory and cellular defects in the pathogenesis of SLE. Various inflammatory and cellular markers including oxidative modifications of proteins, lipids, and DNA contribute to immune system dysregulation and trigger an aggressive autoimmune attack through molecular mechanisms like enhanced NETosis, mTOR pathway activation, and imbalanced T-cell differentiation. Accordingly, the detection of inflammatory and cellular markers is important for providing an accurate assessment of the extent of oxidative stress. Oxidative stress also reduces DNA methylation, thus allowing the increased expression of affected genes. As a result, pharmacological approaches targeting oxidative stress yield promising results in treating patients with SLE. The purpose of this review is to examine the involvement of oxidative stress in the pathogenesis and management of SLE.

Elucidating the phase behavior of ternary model membrane mixtures containing halogenated cholesterol analogues.

Cellular plasma membranes exhibit nanoscale lateral lipid heterogeneity, a feature that is thought to be central to their functionality. However, only a few biophysical methods are capable of detecting domains at sub-micron length scales. We recently showed that cryogenic electron microscopy (cryo-EM) can directly image phase separation in extruded liposomes due to its ability to resolve the intrinsic thickness and electron density differences of ordered and disordered phases. However, the contrast between these phases is poor compared to conventional fluorescence microscopy and is thus both a limiting factor and a focal point for optimization. Because the fundamental source of contrast is the spatial variation in electron density within the bilayer, lipid modifications aimed at selectively increasing the electron density of one phase might enhance the ability to resolve coexisting phases. To this end, we make use of a model membrane mixture of DPPC/DOPC/cholesterol, in which one hydrogen of the methyl group on the steroid ring of cholesterol is replaced by an electron-rich halogen atom. We use a variety of biophysical techniques including confocal fluorescence microscopy, FRET, and cryo-EM to examine phase behavior as a function of composition and temperature. We find that cryo-EM image contrast is increased in liposomes containing halogenated cholesterol as expected. However, the phase behavior of these mixtures is altered compared to mixtures containing native cholesterol providing a cautionary note that halogenated cholesterol has biophysical effects beyond increasing electron density.

Quantification of Dolichyl Phosphates Using Phosphate Methylation and Reverse-Phase Liquid Chromatography-High Resolution Mass Spectrometry.

Dolichyl monophosphates (DolPs) are essential lipids in glycosylation pathways that are highly conserved across almost all domains of life. The availability of DolP is critical for all glycosylation processes, as these lipids serve as membrane-anchored building blocks used by various types of glycosyltransferases to generate complex post-translational modifications of proteins and lipids. The analysis of DolP species by reverse-phase liquid chromatography-mass spectrometry (RPLC-MS) remains a challenge due to their very low abundance and wide range of lipophilicities. Until now, a method for the simultaneous qualitative and quantitative assessment of DolP species from biological membranes has been lacking. Here, we describe a novel approach based on simple sample preparation, rapid and efficient trimethylsilyl diazomethane-dependent phosphate methylation, and RPLC-MS analysis for quantification of DolP species with different isoprene chain lengths. We used this workflow to selectively quantify DolP species from lipid extracts derived of Saccharomyces cerevisiae, HeLa, and human skin fibroblasts from steroid 5-α-reductase 3- congenital disorders of glycosylation (SRD5A3-CDG) patients and healthy controls. Integration of this workflow with global lipidomics analyses will be a powerful tool to expand our understanding of the role of DolPs in pathophysiological alterations of metabolic pathways downstream of HMG-CoA reductase, associated with CDGs, hypercholesterolemia, neurodegeneration, and cancer.

CellPalmSeq: A curated RNAseq database of palmitoylating and de-palmitoylating enzyme expression in human cell types and laboratory cell lines.

The reversible lipid modification protein S-palmitoylation can dynamically modify the localization, diffusion, function, conformation and physical interactions of substrate proteins. Dysregulated S-palmitoylation is associated with a multitude of human diseases including brain and metabolic disorders, viral infection and cancer. However, the diverse expression patterns of the genes that regulate palmitoylation in the broad range of human cell types are currently unexplored, and their expression in commonly used cell lines that are the workhorse of basic and preclinical research are often overlooked when studying palmitoylation dependent processes. We therefore created CellPalmSeq (https://cellpalmseq.med.ubc.ca), a curated RNAseq database and interactive webtool for visualization of the expression patterns of the genes that regulate palmitoylation across human single cell types, bulk tissue, cancer cell lines and commonly used laboratory non-human cell lines. This resource will allow exploration of these expression patterns, revealing important insights into cellular physiology and disease, and will aid with cell line selection and the interpretation of results when studying important cellular processes that depend on protein S-palmitoylation.

Dynamics of CLIMP-63 S-acylation control ER morphology

The complex architecture of the endoplasmic reticulum (ER) comprises distinct dynamic features, many at the nanoscale, that enable the coexistence of the nuclear envelope, regions of dense sheets and a branched tubular network that spans the cytoplasm. A key player in the formation of ER sheets is cytoskeleton-linking membrane protein 63 (CLIMP-63). The mechanisms by which CLIMP-63 coordinates ER structure remain elusive. Here, we address the impact of S-acylation, a reversible post-translational lipid modification, on CLIMP-63 cellular distribution and function. Combining native mass-spectrometry, with kinetic analysis of acylation and deacylation, and data-driven mathematical modelling, we obtain in-depth understanding of the CLIMP-63 life cycle. In the ER, it assembles into trimeric units. These occasionally exit the ER to reach the plasma membrane. However, the majority undergoes S-acylation by ZDHHC6 in the ER where they further assemble into highly stable super-complexes. Using super-resolution microscopy and focused ion beam electron microscopy, we show that CLIMP-63 acylation-deacylation controls the abundance and fenestration of ER sheets. Overall, this study uncovers a dynamic lipid post-translational regulation of ER architecture.

Are Phosphatidic Acids Ubiquitous in Mammalian Tissues or Overemphasized in Mass Spectrometry Imaging Applications?

AbstractMass spectrometry imaging (MSI) is an invaluable tool for the spatial visualization of molecules in vivo. However, the question of whether observed annotations are endogenous or artificial (i. e., from in‐source fragmentation) is critical and has been largely unexplored in multimodal MSI. In matrix‐assisted laser desorption/ionization (MALDI)‐MSI datasets from researchers worldwide, PAs were found to represent up to 18 % of annotations in rat brain. Rat brain was additionally imaged here using nanospray desorption electrospray ionization (nano‐DESI), a softer ionization strategy. No PAs observed with MALDI were present in the nano‐DESI dataset. Further investigation strongly indicated lipid fragmentation to PAs for MALDI‐MSI, but not with nano‐DESI‐MSI. We finally extend this observation to the MALDI‐MSI analyses of human tissues, showing that PA annotations comprised up to 16 % of annotations. Therefore, this study shows that MSI annotations should be carefully interrogated, as in‐source fragmentation or modification of lipids may contribute substantially to false annotations and incorrect biological interpretations.

Lipidation of small GTPase Cdc42 as regulator of its physiological and pathophysiological functions.

The protein cell division cycle 42 (Cdc42) is a small GTPase of the Rho family regulating a plethora of physiological functions in a tissue, cell and subcellular-specific manner via participating in multiple signaling pathways. Since the corresponding signaling hubs are mainly organized along the cellular membranes, cytosolic proteins like Cdc42 need to be properly targeted and held at the membrane. Here, lipid modifications come into play: Cdc42 can be associated with membranes by different lipid anchors including prenylation (Cdc42-prenyl) and palmitoylation (Cdc42-palm). While Cdc42-prenyl is ubiquitously expressed, Cdc42-palm splicing variant in mainly expressed in the brain. Mechanisms underlying Cdc42 lipidation as well as its regulation are the main topic of this review. Furthermore, we will discuss the functional importance of Cdc42 lipid modifications with the focus on the role of different lipids in regulating defined Cdc42 functions. Finally, we will provide an overview of the possible implementation of Cdc42 lipidation in pathological conditions and different diseases.

Investigating sex-specific associations of lipid traits with type 2 diabetes, glycemic traits and sex hormones using Mendelian randomization

BackgroundLow-density lipoprotein (LDL)-cholesterol is positively associated with cardiovascular disease (CVD) and inversely associated with type 2 diabetes, which could detract from lipid modification. Here, we examined whether lipid traits potentially relevant to CVD aetiology, i.e. apolipoprotein B (apoB), triglycerides (TG) and lipoprotein(a) [Lp(a)] exhibited the same associations. We investigated sex-specifically, including the role of sex hormones, because sex disparities exist in lipid profile and type 2 diabetes. We also replicated where possible.MethodsWe used Mendelian randomization (MR) to examine sex-specific associations of apoB, TG and Lp(a) with type 2 diabetes, HbA1c, fasting insulin, fasting glucose, testosterone and estradiol in the largest relevant sex-specific genome-wide association studies (GWAS) in people of European ancestry and replicated where possible. We also assessed sex-specific associations of liability to type 2 diabetes with apoB, TG and Lp(a).ResultsGenetically predicted apoB and Lp(a) had little association with type 2 diabetes or glycemic traits in women or men. Genetically predicted higher TG was associated with higher type 2 diabetes risk [odds ratio (OR) 1.44 per standard deviation (SD), 95% confidence interval (CI) 1.26 to 1.65], HbA1c and fasting insulin specifically in women. Higher TG was associated with lower testosterone in women and higher testosterone in men, but with lower estradiol in men and women. Genetic liability to type 2 diabetes was associated with higher TG in women, and possibly with lower apoB in men.ConclusionsLipid traits potentially relevant to CVD aetiology do not exhibit contrasting associations with CVD and type 2 diabetes. However, higher TG is associated with higher type 2 diabetes risk and glycemic traits, which in turn further increases TG specifically in women, possibly driven by sex hormones.

Membrane targeting with palmitoylated lysine added to PP1-disrupting peptide induces PP1-independent signaling.

Protein phosphatase-1 (PP1) is a ubiquitous enzyme involved in multiple processes inside cells. PP1-disrupting peptides (PDPs) are chemical tools that selectively bind to PP1 and release its activity. To restrict the activity of PDPs to a cellular compartment, we developed PDP-Mem, a cell membrane-targeting PDP. The membrane localization was achieved through the introduction of a palmitoylated lysine. PDP-Mem was shown to activate PP1α in vitro and to localize to the membrane of HeLa Kyoto and U2OS cells. However, in cells, the combination of the polybasic sequence for cell penetration and the membrane targeting palmitoylated lysine activates the MAPK signaling pathway and induces cytoplasmic calcium release independently of PP1 activation. Therefore, when targeting peptides to cellular membranes, undesired effects induced by the targeting sequence and lipid modification need to be considered.

Molecular epidemiology and genomic characterization of a plasmid-mediated mcr-10 and blaNDM-1 co-harboring multidrug-resistant Enterobacter asburiae

Colistin is considered as one of the last-resort antimicrobial agents for treating multidrug-resistant bacterial infections. Multidrug-resistant E. asburiae has been increasingly isolated from clinical patients, which posed a great challenge for antibacterial treatment. This study aimed to report a mcr-10 and blaNDM-1 co-carrying E. asburiae clinical isolate 5549 conferred a high-level resistance against colistin. Antibiotic susceptibility testing was performed using the microdilution broth method. Transferability of mcr-10 and blaNDM-1-carrying plasmids were investigated by conjugation experiments. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) was used to identify modifications in lipid A. Whole genome sequencing and phylogenetic analysis between strain 5549 and a total of 301 E. asburiae genomes retrieved from NCBI database were performed. The genetic characteristics of mcr-10 and blaNDM-1-bearing plasmids were also analyzed. Our study indicated that strain 5549 showed extensively antibiotic-resistant trait, including colistin and carbapenem resistance. The mcr-10 and blaNDM-1 were carried by IncFIB/IncFII type p5549_mcr-10 (159417 bp) and IncN type p5549_NDM-1 (63489 bp), respectively. Conjugation assays identified that only the blaNDM-1-carrying plasmid could be successfully transferred to E. coli J53. Interestingly, mcr-10 did not mediate colistin resistance when it was cloned into E. coli DH5α. Mass spectrometry analysis showed the lipid A palmitoylation of the C-lacyl-oxo-acyl chain to the chemical structure of lipid A at m/z 2063 in strain 5549. In summary, this study is the first to report a mcr-10 and blaNDM-1 co-occurrence E. asburiae recovered from China. Our investigation revealed the distribution of different clonal lineage of E. asburiae with epidemiology perspective and the underlying mechanisms of colistin resistance. Active surveillance is necessary to control the further dissemination of multidrug-resistant E. asburiae.