基于二维凝胶电泳和质谱分析微泡菌ALW1代谢三种海带多糖的蛋白质组

Proteomic Analysis of Three Kelp Polysaccharides Metabolized by Microbulbifer sp. ALW1 Based on Two-Dimensional Gel Electrophoresis and Mass Spectrometry

摘要:

为了阐明海带降解微泡菌ALW1代谢海带多糖的分子机理，本研究进行ALW1菌株代谢海带多糖的蛋白质组研究，利用二维凝胶电泳（2-DE）和质谱（MS）技术鉴定菌株ALW1代谢三种多糖（昆布多糖、岩藻多糖和果胶）的差异蛋白质。二维凝胶电泳显示了202个差异表达的蛋白质斑点，利用质谱最终鉴定了161个蛋白质。聚类分析结果表明，以昆布多糖为碳源培养的细胞内蛋白，与岩藻多糖和果胶培养的细胞内蛋白相比，共鉴定出35种上调蛋白；以岩藻多糖为碳源培养的细胞内蛋白与其他两种蛋白样品相比，共鉴定出88种上调蛋白；以果胶为碳源培养的细胞内蛋白与其他两种蛋白样品相比，共鉴定出68种上调蛋白。GO分析表明，差异表达蛋白质在细胞成分上主要集中在细胞和细胞组成部分，在分子功能上主要集中在催化活性和结合功能上，并且主要参与细胞过程和生物体中的代谢过程。微泡菌ALW1代谢不同海带多糖的蛋白质组研究为该菌株代谢多糖的分子机理，以及利用该菌株生产海带低聚糖和寡糖奠定理论基础。

Abstract:

In order to elucidate the molecular mechanism of kelp polysaccharides metabolism by Microbulbifer sp. ALW1 with kelp-degrading ability, the proteome of kelp polysaccharides metabolism by this strain was studied in this study. Two-dimensional gel electrophoresis (2-DE) and mass spectrometry (MS) were utilized to identify the differential proteins of strain ALW1 metabolizing three kelp polysaccharides (laminarin, fucoidan, and pectin). The results of two-dimensional gel electrophoresis revealed 202 differentially expressed protein spots, and 161 proteins were finally identified using mass spectrometry. Cluster analysis showed that a total of 35 up-regulated proteins were identified in intracellular proteins cultured with laminarin as a carbon source compared to those cultured with fucoidan and pectin. A total of 88 up-regulated proteins were identified in intracellular proteins cultured with fucoidan as a carbon source compared to the other two protein samples. A total of 68 up-regulated proteins were identified in intracellular proteins cultured with pectin as a carbon source compared to the other two protein samples. GO analysis showed that differentially expressed proteins were mainly concentrated in cell and cell part in terms of cellular component, mainly concentrated in catalytic activity and binding function in terms of molecular function, and mainly involved in cellular process and metabolic process in terms of biological process. The proteomic study on the metabolism of different kelp polysaccharides by Microbulbifer sp. ALW1 lays a theoretical foundation for the molecular mechanism of polysaccharides metabolism by this strain and the production of kelp low molecular weight polysaccharides and oligosaccharides using this strain.