Proteomic analysis of stachyose contribution to the growth of Lactobacillus acidophilus CICC22162
Proteomic analysis of stachyose contribution to the growth of Lactobacillus acidophilus CICC22162
复制标题
水苏糖对嗜酸乳杆菌 CICC22162 生长的贡献的蛋白质组学分析
DOI：
10.1039/c8fo00528a
复制
发表时间：
2018
期刊：
影响因子：
6.1
通讯作者：
Huang Wei zhi
中科院分区：
文献类型：
--
作者：
Zhong Xian feng;Zhang Yu bo;Huang Gui dong;Ouyang Yong zhong;Liao Dong jiang;Peng Jia wei;Huang Wei zhi
水苏糖是一种功能性低聚糖，是一种潜在的结肠发酵益生元。为了了解水苏糖促进益生菌生长的机制，我们分析了在水苏糖和葡萄糖条件下生长的嗜酸乳杆菌蛋白质组的差异。通过双向电泳和质谱分析相结合，我们在这两种条件下观察到16种蛋白质的差异丰度，并鉴定出9个蛋白质斑点。当水苏糖作为唯一碳源时，其中6种蛋白质含量很高。它们包括磷酸转移酶系统、能量偶联因子（ECF）转运体和甘露糖-6-磷酸异构酶，它们参与嗜酸乳杆菌CICC22162中水苏糖的摄取和分解代谢。这些观察结果被定量RT-PCR分析和酶活性测定所证实。蛋白质含量与其mRNA水平呈正相关。此外，我们通过MD模拟和自由能分析探索了水苏糖与新鉴定的ECF转运体结合的识别机制。综上所述，这些结果为水苏糖促进益生菌生长的机制提供了新的见解。
Stachyose is a functional oligosaccharide, acting as a potential prebiotic for colonic fermentation. To understand the mechanism of how stachyose promotes the growth of probiotic bacterium, we analyzed the differences of the proteome of Lactobacillus acidophilus grown on stachyose or glucose. By a combination of two-dimensional electrophoresis and mass spectrometry analysis, we observed 16 proteins differentially abundant under these two conditions and identified 9 protein spots. Six of these proteins were highly abundant when stachyose was used as the sole carbon source. They included the phosphotransferase system, the energy coupling factor (ECF) transporter and the mannose-6-phosphate isomerase, involved in the uptake and catabolism of stachyose in Lactobacillus acidophilus CICC22162. Supportively, these observations were validated by quantitative RT-PCR analysis and enzymatic activity determination. Positive correlation was found between the content of the proteins and their mRNA levels. Additionally, we explored the recognition mechanism for stachyose binding to the newly identified ECF transporter by MD simulations and free energy analysis. Taken together, these results provide new insights into the mechanism of stachyose in promoting the growth of probiotic bacterium.