Optimization of Protein Quantification From Soil Samples

IF 3.8 2区 农林科学 Q2 SOIL SCIENCE
M. Waibel, M. Tuohy, E. Paterson, B. Thornton, F. Brennan, F. Abram
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引用次数: 0

Abstract

Protein profiling of soil samples has the potential to enhance our understanding of soil ecosystems and guide the development of sustainable soil management practices. In that context, there is a need to develop robust proteomic workflows, starting with reliable protein quantification. Total protein quantification with the Lowry assay is a relatively easy, rapid, and cheap method, but requires interference corrections due to reactivity with a wide range of compounds that occur in soil, plant, and other biological matrices. Here, we propose sample-specific corrections for soil protein extracts. We benchmarked our approach against other protein quantification methods, including other Lowry corrections, total hydrolysable amino acids, and Qubit total protein assay. Our sample-specific Lowry corrections did not overestimate or underestimate protein content when compared to the other methods tested. As a practical contribution, this work provides a calibration method for the Lowry assay using protein reference values in a multivariate regression approach to enable simple and high-throughput total protein quantification of soil samples.

Abstract Image

土壤样品中蛋白质定量的优化
土壤样品的蛋白质分析有可能增强我们对土壤生态系统的理解,并指导可持续土壤管理实践的发展。在这种情况下,需要从可靠的蛋白质定量开始,开发强大的蛋白质组学工作流程。用Lowry法定量总蛋白是一种相对简单、快速和廉价的方法,但由于与土壤、植物和其他生物基质中出现的各种化合物的反应性,需要进行干扰校正。在这里,我们提出了土壤蛋白质提取物的样品特异性校正。我们将我们的方法与其他蛋白质定量方法进行了基准比较,包括其他Lowry校正,总水解氨基酸和Qubit总蛋白测定。与其他测试方法相比,我们的样品特异性Lowry校正没有高估或低估蛋白质含量。作为一个实际的贡献,这项工作提供了一种校正方法,在多元回归方法中使用蛋白质参考值,以实现土壤样品的简单和高通量总蛋白质定量。
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来源期刊
European Journal of Soil Science
European Journal of Soil Science 农林科学-土壤科学
CiteScore
8.20
自引率
4.80%
发文量
117
审稿时长
5 months
期刊介绍: The EJSS is an international journal that publishes outstanding papers in soil science that advance the theoretical and mechanistic understanding of physical, chemical and biological processes and their interactions in soils acting from molecular to continental scales in natural and managed environments.
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