Environmental Biotechnology Institute, Environmental Science Program, University of Idaho, Moscow, Idaho 83844, USA.
Appl Environ Microbiol. 2011 Feb;77(3):810-20. doi: 10.1128/AEM.01323-10. Epub 2010 Nov 29.
Perchlorate is a known health hazard for humans, fish, and other species. Therefore, it is important to assess the response of an ecosystem exposed to perchlorate contamination. The data reported here show that a liquid chromatography-mass spectrometry-based proteomics approach for the detection of perchlorate-reducing enzymes can be used to measure the ability of microorganisms to degrade perchlorate, including determining the current perchlorate degradation status. Signature peptides derived from chlorite dismutase (CD) and perchlorate reductase can be used as biomarkers of perchlorate presence and biodegradation. Four peptides each derived from CD and perchlorate reductase subunit A (PcrA) and seven peptides derived from perchlorate reductase subunit B (PcrB) were identified as signature biomarkers for perchlorate degradation, as these sequences are conserved in the majority of the pure and mixed perchlorate-degrading microbial cultures examined. However, chlorite dismutase signature biomarker peptides from Dechloromonas agitata CKB were found to be different from those in other cultures used and should also be included with selected CD biomarkers. The combination of these peptides derived from the two enzymes represents a promising perchlorate presence/biodegradation biomarker system. The biomarker peptides were detected at perchlorate concentrations as low as 0.1 mM and at different time points both in pure cultures and within perchlorate-reducing environmental enrichment consortia. The peptide biomarkers were also detected in the simultaneous presence of perchlorate and an alternate electron acceptor, nitrate. We believe that this technique can be useful for monitoring bioremediation processes for other anthropogenic environmental contaminants with known metabolic pathways.
高氯酸盐对人类、鱼类和其他物种是一种已知的健康危害。因此,评估暴露于高氯酸盐污染的生态系统的反应非常重要。这里报告的数据表明,基于液相色谱-质谱的蛋白质组学方法可用于检测高氯酸盐还原酶,以测量微生物降解高氯酸盐的能力,包括确定当前高氯酸盐的降解状态。来源于亚氯酸盐歧化酶(CD)和高氯酸盐还原酶的特征肽可作为高氯酸盐存在和生物降解的生物标志物。从 CD 和高氯酸盐还原酶亚基 A(PcrA)各衍生出 4 个肽,从高氯酸盐还原酶亚基 B(PcrB)衍生出 7 个肽,被鉴定为高氯酸盐降解的特征生物标志物,因为这些序列在大多数经过测试的纯和混合高氯酸盐降解微生物培养物中都保守。然而,Dechloromonas agitata CKB 的亚氯酸盐歧化酶特征生物标志物肽与其他培养物中的肽不同,也应该与选定的 CD 生物标志物一起包含。这两种酶的生物标志物肽的组合代表了一种有前途的高氯酸盐存在/生物降解生物标志物系统。这些肽生物标志物在低至 0.1mM 的高氯酸盐浓度和纯培养物以及高氯酸盐还原环境富集联合体中的不同时间点都可以检测到。在同时存在高氯酸盐和替代电子受体硝酸盐的情况下,也检测到了这些肽生物标志物。我们相信,这项技术对于监测其他具有已知代谢途径的人为环境污染物的生物修复过程非常有用。