Extraction of microbial proteome from soil: potential and limitations assessed through a model study
Extraction of microbial proteome from soil: potential and limitations assessed through a model study
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DOI：
10.1111/j.1365-2389.2010.01322.x
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发表时间：
2011-02-01
影响因子：
4.2
通讯作者：
Renella, G.
中科院分区：
文献类型：
--
作者：
Giagnoni, L.;Magherini, F.;Renella, G.
蛋白质组学是基于蛋白质表达谱分析来研究生物系统的功能和调控的学科，人们普遍认为土壤蛋白质组学可能是更好地进行土壤管理的工具。由于土壤通过各种机制稳定胞外蛋白的能力，土壤蛋白质组学的发展需要评估从各种土壤类型中提取蛋白质的效率。我们评估了由Cupriavidus metallidurans CH 34提取土壤微生物蛋白质组的可能性，它具有已知的蛋白质组，到无菌砂，高岭石，蒙脱石和砂，高岭石，蒙脱石，针铁矿和腐殖酸的混合物。接种后1h，C.通过菌落形成单位法（CFU）测定金属硬聚糖的量，通过Bradford法测定提取的蛋白质的量，并且通过二维凝胶电泳技术（2D-GE）分析细菌蛋白质组。细菌数量为2.5 × 106 CFU g-1的土壤中的所有缩影，而总提取的蛋白质含量从98.1到1268 μ g g-1的各种缩影中的变化，但在接种蒙脱石检测不到。从细菌培养物和接种的缩影的蛋白质斑点的数量变化之间的317和591，与54个变异点之间的纯培养物和缩影。在蒙脱石微观世界的2D-GE中未检测到蛋白质斑点。人工土壤微宇宙的2D-GE显示出与纯培养物以及砂和高岭石微宇宙不同的蛋白质模式。结果证实了粘土比表面积和CEC在蛋白质吸附中的重要性，因为蒙脱石单独具有最大的吸附容量，并且表明所使用的人工土壤也具有对微生物蛋白质的大吸附容量。在全球范围内，研究结果表明，从土壤中提取蛋白质的强烈影响的粘土类型和有机质含量，蛋白质提取效率差，可能会降低土壤蛋白质组学的潜力。
Proteomics is the study of functions and regulation of biological systems based on the analysis of the protein expression profile, and there is a general agreement that soil proteomics may be a tool for better soil management. Because of the ability of soils to stabilize extracellular proteins by various mechanisms, development of soil proteomics needs an assessment of the efficiency of protein extraction from various soil types. We evaluated the possibility of extraction of soil microbial proteome by inoculating Cupriavidus metallidurans CH34, which has a known proteome, into sterile sand, kaolinite, montmorillonite and a mixture of sand, kaolinite, montmorillonite, goethite and humic acids. One hour after inoculation, the viability of C. metallidurans was determined by the colony-forming units method (CFU), the amount of extracted proteins was determined by the Bradford method and the bacterial proteome was analysed by the two-dimensional gel electrophoresis technique (2D-GE). The bacterial number was 2.5 x 106 CFU g-1 of soil in all microcosms, whereas the total extracted protein content varied from 98.1 to 1268 mu g g-1 in the various microcosms, but was undetectable in the inoculated montmorillonite. The number of protein spots from the bacterial culture and the inoculated microcosms varied between 317 and 591, with 54 variable spots among the pure culture and the microcosms. No protein spots were detected in the 2D-GE from the montmorillonite microcosm. The 2D-GE of artificial soil microcosms showed a protein pattern that was different from those of pure culture and sand and kaolinite microcosms. The results confirm the importance of clay-specific surface area and CEC in protein adsorption as montmorillonite alone had the largest sorptive capacity, and show that the artificial soil used also had a large sorptive capacity for microbial proteins. Globally, the results indicate that the extraction of proteins from soils is strongly influenced by the clay type and organic matter content, and that poor protein extraction efficiency may reduce the potential of soil proteomics.