Evaluation of simplified ester-linked fatty acid analysis (ELFA) for phospholipid fatty acid (PLFA) analysis of bacterial population

IF 2.6 4区生物学 Q2 BIOCHEMICAL RESEARCH METHODS Analytical biochemistry Pub Date : 2024-08-08 DOI:10.1016/j.ab.2024.115638

Jeong Hyeon Hwang , Tae-Rim Choi , Suwon Kim , Yeda Lee , Yuni Shin , Suhye Choi , Jinok Oh , Sang-Hyoun Kim , Jeong-Hoon Park , Shashi Kant Bhatia , Yung-Hun Yang

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Abstract

Phospholipid fatty acid (PLFA) analysis is used for characterizing microbial communities based on their lipid profiles. This method avoids biases from PCR or culture, allowing data collection in a natural state. However, PLFA is labor-intensive due to lipid fractionation. Simplified ester-linked fatty acid analysis (ELFA), which skips lipid fractionation, offers an alternative. It utilizes base-catalyzed methylation to derivatize only lipids, not free fatty acids, and found glycolipid and neutral lipid fractions are scarcely present in most bacteria, allowing lipid fractionation to be skipped. ELFA method showed a high correlation to PLFA data (r = 0.99) and higher sensitivity than the PLFA method by 1.5–2.57-fold, mainly due to the higher recovery of lipids, which was 1.5–1.9 times higher than with PLFA. The theoretical limit of detection (LOD) and limit of quantification (LOQ) for the ELFA method indicated that 1.54-fold less sample was needed for analysis than with the PLFA method. Our analysis of three bacterial cultures and a simulated consortium revealed the effectiveness of the ELFA method by its simple procedure and enhanced sensitivity for detecting strain-specific markers, which were not detected in PLFA analysis. Overall, this method could be easily used for the population analysis of synthetic consortia.

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评估用于细菌群体磷脂脂肪酸 (PLFA) 分析的简化酯联脂肪酸分析法 (ELFA)。

磷脂脂肪酸（PLFA）分析用于根据微生物群落的脂质特征描述其特性。这种方法避免了 PCR 或培养产生的偏差，可在自然状态下收集数据。然而，由于脂质分馏，PLFA 需要大量人力。简化的酯联脂肪酸分析（ELFA）可省去脂质分馏，提供了一种替代方法。它利用碱催化甲基化只衍生脂质，而不衍生游离脂肪酸，发现大多数细菌中几乎不存在糖脂和中性脂质部分，因此可以跳过脂质分馏。ELFA 方法与 PLFA 数据的相关性很高（r=0.99），灵敏度比 PLFA 方法高 1.5-2.57 倍，主要原因是脂质的回收率更高，是 PLFA 方法的 1.5-1.9 倍。ELFA 方法的理论检测限（LOD）和定量限（LOQ）表明，与 PLFA 方法相比，分析所需的样品量减少了 1.54 倍。我们对三种细菌培养物和一个模拟菌群进行的分析表明，ELFA 方法操作简单，对检测菌株特异性标记物的灵敏度更高（PLFA 分析中检测不到这些标记物），因此非常有效。总之，这种方法可轻松用于合成菌群的群体分析。

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来源期刊

Analytical biochemistry 生物-分析化学

CiteScore

5.70

自引率

0.00%

发文量

283

审稿时长

44 days

期刊介绍： The journal''s title Analytical Biochemistry: Methods in the Biological Sciences declares its broad scope: methods for the basic biological sciences that include biochemistry, molecular genetics, cell biology, proteomics, immunology, bioinformatics and wherever the frontiers of research take the field. The emphasis is on methods from the strictly analytical to the more preparative that would include novel approaches to protein purification as well as improvements in cell and organ culture. The actual techniques are equally inclusive ranging from aptamers to zymology. The journal has been particularly active in: -Analytical techniques for biological molecules- Aptamer selection and utilization- Biosensors- Chromatography- Cloning, sequencing and mutagenesis- Electrochemical methods- Electrophoresis- Enzyme characterization methods- Immunological approaches- Mass spectrometry of proteins and nucleic acids- Metabolomics- Nano level techniques- Optical spectroscopy in all its forms. The journal is reluctant to include most drug and strictly clinical studies as there are more suitable publication platforms for these types of papers.