基于液相色谱-质谱的多肽组学技术用于鉴定紫芝菌丝体胞内外的多肽组成  

Identification of intracellular and extracellular polypeptide composition of Ganoderma sinense mycelium using liquid chromatography-mass spectrometry based peptidomic approaches

作　　者：伍强 王正伟 肖珍艳 WU Qiang;WANG Zhengwei;XIAO Zhenyan(School of Food and Chemical Engineering,Shaoyang University,Shaoyang 422000,China;Hunan Provincial Key Laboratory of Forestry Biotechnology,Central South University of Forestry and Technology,Changsha 410004,China)

机构地区：[1]邵阳学院食品与化学工程学院,湖南邵阳422000 [2]中南林业科技大学林业生物技术湖南省重点实验室,湖南长沙410004

出　　处：《邵阳学院学报(自然科学版)》2025年第2期58-66,共9页Journal of Shaoyang University:Natural Science Edition

基　　金：国家自然科学基金青年项目(32202573);湖南省教育厅科学研究重点项目(22A0538);湖南省研究生科研创新项目(CX20231305)。

摘　　要：为揭示紫芝菌丝体胞内外的多肽组成差异,通过超滤膜与TSR-gel G2500 PWXL柱有效分离出多肽组分,进一步利用液相色谱-质谱组学技术分析紫芝胞内外的多肽组成差异与代谢特征。Venn图分析表明,紫芝胞内特有881种多肽,胞外特有146种,582种为胞内外共有,显示胞内多肽种类更为丰富。主成分分析(principal component andlysis,PCA)分析显示,紫芝胞内外多肽在主成分维度上明显分离,反映了胞内代谢的高度组织化及胞外代谢的多样性。此外,胞内总离子流图(total ion chromatogram,TIC)显示多个高丰度离子峰,表明其化学复杂性高于胞外。火山图和热图分析进一步显示,紫芝胞内多肽的表达水平显著高于胞外,尤其在脂肪酰基和肽代谢途径中表现活跃。紫芝胞内代谢物的KO富集结果集中于蛋白质合成、氧化磷酸化及氨基酸代谢等基础通路,而胞外代谢物则与植物-病原体相互作用及酒精代谢等功能相关。本研究为解析紫芝胞内外的多肽组成与代谢差异并进一步开发其生物活性物质提供了较重要依据。This study focused on the compositional difference of intracellular and extracellular peptides of Ganoderma sinense(G.sinense)mycelium.Using ultrafiltration membrane and TSR-gel G2500 PWXL column,peptide compositions were effectively separated out,followed by liquid chromatography-mass spectrometry(LC-MS)analysis to verify differences in peptide composition and metabolic characteristics between intracellular and extracellular peptides in G.sinense mycelium.Venn diagram analysis identified 881 peptides unique to intracellular fractions,146 to extracellular fractions,and 582 shared,indicating a higher diversity of peptides in intracellular fractions.A clear separation between intracellular and extracellular peptides was observed in principal component analysis(PCA),reflecting the highly organized nature of intracellular metabolism and the diversity of extracellular metabolism.Additionally,intracellular total ion chromatogram(TIC)profiles exhibited numerous high-abundance ion peaks,highlighting greater chemical complexity than extracellular profiles.Volcano and heatmap analysis further revealed significantly higher expression levels of intracellular peptides,particularly those involved in fatty acyl and peptide metabolic pathways.Furthermore,KO enrichment analysis showed that intracellular metabolites were associated with fundamental pathways,such as protein synthesis,oxidative phosphorylation,and amino acid metabolism,while extracellular metabolites were enriched in pathways related to plant-pathogen interactions and alcohol metabolism.This study provided important insights into the compositional difference and metabolic characteristics of G.sinense mycelium and offers a valuable foundation for developing its bioactive compounds.

关 键 词：紫芝 菌丝体 多肽 多肽组学 

分 类 号：Q936[生物学—微生物学]