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甲基转移酶:酸性土壤和沉积物中微生物甲基汞形成的酶活性测定

Methyltransferase: an enzyme assay for microbial methylmercury formation in acidic soils and sediments.

作者信息

Siciliano Steven D, Lean David R S

机构信息

Department of Biology, University of Ottawa, Ontario, Canada.

出版信息

Environ Toxicol Chem. 2002 Jun;21(6):1184-90.

PMID:12069301
Abstract

Methylmercury (MeHg) is a highly toxic form of mercury that bioaccumulates in aquatic food chains. However, methods to reliably identify sites of MeHg formation or to quantify MeHg production require the use of isotopic tracers, which limits their use. In this paper, a method is presented to quantify the methylation of mercury by a methyltransferase pathway. This methyltransferase pathway is one of the biochemical pathways responsible for biological mercury methylation. Protein is extracted from environmental samples, and mercury methyltransferase (HgMT) activity of soil extracts is calculated by assessing increases in methyltransferase activity induced by Hg addition. In enzyme extracts from pure cultures or soil samples, HgMT activity correlated with net MeHg production and Hg consumption, suggesting that HgMT activity can be used to estimate MeHg production in field samples. Over the course of a three-month period in a freshwater wetland, HgMT activity correlated with net MeHg concentrations (r2 = 0.55; p < 0.057). Furthermore, HgMT activity predicted (r2 = 0.80; p < 0.01) gross MeHg formation in freshwater wetlands as well as in laboratory microcosms calculated using previously published rate constants. Our results show that a methyltransferase assay, in combination with demethylation estimates, accurately predicts MeHg formation under field and laboratory conditions. This assay does not require the use of mercury added to field samples to estimate activity but rather estimates the biological activity present in the soil by quantifying the amount of enzyme present in the soil. Such an assay is well suited for use in field surveillance programs assessing MeHg formation in a variety of environments.

摘要

甲基汞(MeHg)是汞的一种剧毒形式,会在水生食物链中生物累积。然而,可靠识别MeHg形成位点或量化MeHg生成量的方法需要使用同位素示踪剂,这限制了它们的应用。本文提出了一种通过甲基转移酶途径量化汞甲基化的方法。这种甲基转移酶途径是负责生物汞甲基化的生化途径之一。从环境样品中提取蛋白质,并通过评估添加汞后诱导的甲基转移酶活性增加来计算土壤提取物的汞甲基转移酶(HgMT)活性。在纯培养物或土壤样品的酶提取物中,HgMT活性与净MeHg生成量和汞消耗量相关,这表明HgMT活性可用于估计野外样品中的MeHg生成量。在一个淡水湿地为期三个月的时间里,HgMT活性与净MeHg浓度相关(r2 = 0.55;p < 0.057)。此外,HgMT活性预测了(r2 = 0.80;p < 0.01)淡水湿地以及使用先前公布的速率常数计算的实验室微观世界中的总MeHg形成量。我们的结果表明,甲基转移酶测定法与去甲基化估计相结合,能够准确预测野外和实验室条件下的MeHg形成。该测定法不需要使用添加到野外样品中的汞来估计活性,而是通过量化土壤中存在的酶量来估计土壤中存在的生物活性。这样的测定法非常适合用于评估各种环境中MeHg形成的野外监测计划。

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Methyltransferase: an enzyme assay for microbial methylmercury formation in acidic soils and sediments.甲基转移酶:酸性土壤和沉积物中微生物甲基汞形成的酶活性测定
Environ Toxicol Chem. 2002 Jun;21(6):1184-90.
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