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Lipids are a class of metabolites with diverse chemical structures. Lipids are not only components of cell membranes and energy storage substances, but also perform a variety of important biological functions in life activities. As a branch of metabolomics, lipidomics is a discipline that systematically studies lipids and their interacting molecules in organisms, tissues or cells. Microbial lipidomics can help with phenotypic studies, infectiousness studies, drug resistance mechanism studies, and studies of lipids and their interacting molecules in microorganisms.

Applications of lipidomics in microbiology

  • Biomarker discovery

A variety of stress responses in microorganisms are accompanied by disordered lipid metabolism, so the search for lipid biomarkers is of great importance for microbial studies. For example, microbial fermentation produces ethanol, which is a renewable energy source that can be used to cope with the energy crisis we are currently facing. However, the process is inhibited by furans, phenols and acetic acid produced by Saccharomyces cerevisiae during the fermentation process. Comparative lipidomic studies of parental (SC) and furan-resistant (SCF), phenol-resistant (SCP), and acetic acid-resistant (SCA) bacteria revealed that phosphatidylcholine, phosphatidylinositol, and phosphatidic acid distinguish SC from SCF, SCP, and SCA, respectively, and are biomarkers of these three resistant bacteria.

  • Search for drug targets and new drug development

The study of lipid metabolites and related enzymes as a target to find new drug targets plays an important role in new drug development. For example, the formation of the periplasm of Candida parapsilosis is accompanied by the upregulation of FAs2 gene when the environmental oxygen content is low. In response to this phenomenon, some scholars observed the growth of FAs2 knockout bacteria with exogenous addition of different long-chain FAs, and the results showed that the virulence of the mutant bacteria was significantly reduced, and they could not produce unsaturated FAs, form normal biofilm, and be killed by macrophages. This suggests that FAs may be potential therapeutic targets. Lipidomics can provide information on the changes of microbial lipids and related enzymes, which can help to elucidate microbial-related mechanisms of action and play an important role in microbiology research.

  • Optimization of fermentation conditions

Ethanol production during fermentation is important for the efficient production of fuel alcohol, wine, and other alcoholic beverages. However, increasing ethanol concentration can slow down the conversion of sugars to ethanol. A study was conducted to evaluate the relationship between lipids and fermentation kinetic parameters by performing lipidomic studies on 22 industrial brewer's yeasts with different ethanol tolerance. The results showed that lipid composition was related to ethanol concentration as an indicator of ethanol tolerance of the strains and that the maximum cell concentration affecting ethanol tolerance was also related to lipids. Among them, strains with low ethanol production were associated with higher levels of phosphatidylinositol in the early stages of fermentation, while strains with higher cell concentration and ethanol production were associated with phosphatidylcholine.

Lipidomics in Microbiology

Schematic overview of the lipidomic data process and analysis workflow (Ding et al., 2021).

Lipidomics research methods in microbiology

Improvements in mass spectrometry have greatly contributed to the development of lipidomics, especially soft ionization techniques such as matrix-assisted laser desorption/ionization (MALDI) and electrospray ionization (ESI). MALDI is a widely used ionization method for protein research and has been successfully applied to lipid analysis.

The advantages of MALDI are: (1) the higher the solubility of lipids and matrices in organic solvents, the better the signal-to-noise ratio and reproducibility; and (2) compared with ESI, MALDI is more tolerant to salt and is suitable for the analysis of polar lipids, such as phospholipids.

Triple quadrupole (3Q) generates a large amount of molecular structure information and has become a common mass analyzer in lipidomics. It has four scanning modes: product-ion scan, precursor-ion scan, neutral-loss scan and selected reaction monitoring, but the quality accuracy and resolution are not very high. Other mass analyzers, such as time-of-flight mass spectrometry (TOF) and ion-trap mass spectrometry (Ion-Trap) have high accuracy and resolution and can be used to determine the elemental composition of lipid molecules.

Creative Proteomics has a state-of-the-art chromatography-mass spectrometry detection platform to provide professional lipidomics analysis services to our clients.

Reference:

  1. Ding, S., Bale, N. J., et al. (2021). Lipidomics of environmental microbial communities. II: Characterization using molecular networking and information theory. Frontiers in microbiology, 12, 1920.
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Phospholipid molecules are a class of amphiphilic compounds with phosphate groups, containing both a hydrophobic tail consisting of fatty acid chains and a hydrophilic head consisting of substituted groups linked by phosphate. There are seven major groups of phospholipids in plasma: Phosphatidylcholine (PC)Lyso-phosphatidylcholine (Lyso-PC)Phosphatidylethanolamine (PE)Phosphatidylserine (PS)Phosphatidylinositol (PI)Phosphatidylglycerol (PG), and Sphingomyelin (SM).

Functions of phospholipids

Phospholipids not only maintain the normal morphology of cell membranes, but also participate in a large number of vital activities of living organisms.

Phospholipids play an important role in signaling, substance exchange, apoptosis and proliferation, e.g. glycerophospholipids are the precursors of lipid second messengers.

Phospholipid metabolites also play an important role in fever, inflammatory response and platelet aggregation, such as arachidonic acid, prostaglandins and platelet-activating factor.

Studies have shown that phospholipids are important bioactive substances and storage forms of information molecule precursors in the body. Its hydrolysis and oxidation products can regulate human metabolism by affecting the secretory or nervous system, improve memory, etc. It is also a chemical mediator in the formation of many diseases.

In addition, many diseases have been reported to be associated with abnormal phospholipid metabolism, such as diabetes, Alzheimer's disease, pancreatic cancer, etc. Therefore, in recent years, research targeting the structure, function and metabolism of phospholipids has become a hot topic in biology, medicine and pharmacology.

Each phospholipid is not a simple compound. The composition of phospholipids, depending on the polar head groups, the length of carbon chains, and the number of unsaturated bonds, is highly complex and diverse, so the analysis of phospholipids can be very difficult.

Phospholipids: Functions and Analysis

Phospholipid detection methods

Due to the development of soft ionization techniques (electrospray ionization (ESI) and matrix-assisted laser desorption ionization (MALDI)), mass spectrometry detectors are increasingly used in the analysis of phospholipids. The soft ionization technique, which produces almost no fragment ions, is particularly suitable for the identification of the molecular weight of complex compounds. In contrast, mass spectrometry, represented by high-resolution time-of-flight TOF, has powerful qualitative capabilities and is particularly suitable for the identification of complex mixtures of phospholipids.

Although the linear range of ESI in quantitation is not as wide as UV detector, mass spectrometry detection has the advantage of low detection limit and good specificity. Compared to other chromatographic common detectors, when performing quantitative chromatographic analysis, it is necessary to ensure the singularity of the chromatographic peak, and the chromatographic peak must not contain other interfering components. Mass spectrometry is less demanding in this regard, allowing for multiple reaction monitoring, parent ion, daughter ion scanning and neutral loss scanning, making it more suitable for the identification of low concentrations of phospholipids in complex matrices.

Creative Proteomics uses liquid chromatography-electrospray ionization mass spectrometry (LC-ESI/MS) as the core research tool for phospholipid studies, providing a wide range of services for disease diagnosis and new drug development.