A simple cleanup method for the removal of humic substances from soil protein extracts using aluminum coagulation
A simple cleanup method for the removal of humic substances from soil protein extracts using aluminum coagulation
复制标题
DOI：
10.1007/s11356-018-2434-z
复制
发表时间：
2018-08-01
影响因子：
5.8
通讯作者：
Stamatakis, Aristeidis
中科院分区：
文献类型：
--
作者：
Mandalakis, Manolis;Panikov, Nicolai S.;Stamatakis, Aristeidis
土壤蛋白质组学是对土壤中整个蛋白质补体的大规模表征，为破译陆地生态系统中微生物功能的作用提供了一种有前途的方法。然而，提取足够数量和足够纯度的土壤蛋白质仍然是一项具有挑战性的任务，主要是由于干扰腐殖质物质的共同提取。到目前为止，用液体苯酚处理土壤提取物一直是减少腐殖质的“黄金标准”，而NoviPure清理工具包最近推出了一种无毒的方法。本研究描述了一种基于三价铝离子（Al3+）腐殖质凝固的递送高纯度蛋白质的替代方法。为了最大限度地提高蛋白质产量，减少共提取腐殖质，对各实验参数进行了单独优化。将优化后的方法应用于一组具有不同理化特性的土壤样品，并与其他两种方法进行比较。Al3+处理的腐殖物残留量比苯酚处理和NoviPure Kit处理的腐殖物残留量分别高出26%和35%，但差异具有边际统计学意义。对于提取的蛋白质，三种方法的平均产量具有可比性，没有统计学上的显著差异。总的来说，Al3+腐殖质混凝在蛋白质产量和纯度方面提供了相当的清理性能，但它比酚分配方法毒性更小，更简单，而它比NoviPure Kit便宜得多。这项新技术有望促进土壤蛋白质组学研究的实施。
Soil proteomics, the large-scale characterization of the entire protein complement in soils, provides a promising approach for deciphering the role of microbial functioning in terrestrial ecosystems. However, the extraction of soil proteins in sufficient quantities and of adequate purity remains a challenging task mainly due to the co-extraction of interfering humic substances. Up to now, the treatment of soil extracts with liquid phenol has been the "gold standard" for reducing humics, while the NoviPure cleanup kit was recently launched as a non-toxic approach. The present study describes an alternative method for delivering high-purity proteins based on humic coagulation with trivalent aluminum ions (Al3+). Various experimental parameters were optimized individually in order to maximize protein yield and diminish co-extracted humics. The optimized method was applied on a set of soil samples with diverse physicochemical characteristics and a comparison with the other two techniques was conducted. The amount of residual humics resulting from Al3+-based method was 26 and 35% higher than that from phenol treatment and NoviPure Kit, respectively, but these differences were of marginal statistical significance. With regard to extracted proteins, the average yields of the three methods were comparable, without showing any statistically significant differences. Overall, humic coagulation with Al3+ offers comparable cleanup performance in terms of protein yield and purity, but it is less toxic and less complex than the phenol-partitioning method, whereas it is far less expensive than the NoviPure Kit. The new technique is expected to facilitate the implementation of proteomic studies in soils.