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Isolation and Proteomic Analysis of Extracellular Vesicles from Lactobacillus salivarius SNK-6

文献类型： 外文期刊

作者： Huang, Jiwen 1 ; Zhao, Ayong 1 ; He, Daqian 2 ; Wu, Xiao 3 ; Yan, Huaxiang 2 ; Zhu, Lihui 2 ;

作者机构： 1.Zhejiang Agr & Forestry Univ, Coll Anim Sci & Technol, Hangzhou 311300, Peoples R China

2.Shanghai Acad Agr Sci, Inst Anim Husb & Vet Sci, Shanghai 201106, Peoples R China

3.Shanghai Acad Agr Sci, Biotechnol Res Inst, Key Lab Agr Genet & Breeding, Shanghai 201106, Peoples R China

关键词： Extracellular vesicles; Lactobacillus salivarius; protein profile

期刊名称：JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY （ 影响因子：2.8； 五年影响因子：3.0 ）

ISSN： 1017-7825

年卷期： 2024 年 34 卷 1 期

页码：

收录情况： SCI

摘要： The proteins carried by the extracellular vesicles of Lactobacillus salivarius SNK-6 (LsEVs) were identified to provide a foundation for further explorations of the probiotic activities of L. salivarius SNK-6. LsEVs were isolated from the culture media of L. salivarius SNK-6 and morphological analysis was conducted by scanning electron microscopy. Subsequent transmission electron microscopy and nanoparticle tracking analysis were performed to assess the morphology and particle size of the LsEVs. In addition, the protein composition of LsEVs was analyzed using silver staining and protein mass spectrometry. Finally, internalization of the identified LsEVs was confirmed using a confocal microscope, and enzyme-linked immunosorbent assay was employed to analyze the levels of inflammatory cytokines in LPS-challenged RAW264.7 cells. The results revealed that the membraneenclosed LsEVs were spherical, with diameters ranging from 100-250 nm. The LsEVs with diameters of 111-256 nm contained the greatest amount of cargo. In total, 320 proteins (10-38 kD) were identified in the LsEVs and included anti-inflammatory molecules, such as PrtP proteinase, cochaperones, and elongation factor Tu, as well as some proteins involved in glycolysis/gluconeogenesis, such as fructose-1,6-bisphosphate aldolase. Enrichment analysis showed these proteins to be related to the terms "metabolic pathway," "ribosome," "glycolysis/gluconeogenesis," "carbohydrate metabolism," and "amino acid metabolism." Furthermore, the LsEVs were internalized by host liver cells and can regulate inflammation. These findings confirm that LsEVs contain various functional proteins that play important roles in energy metabolism, signal transduction, and biosynthesis.

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