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红树林沉积物中外源化合物生物降解的宏基因组景观。

The metagenomic landscape of xenobiotics biodegradation in mangrove sediments.

机构信息

Center for Chemistry, Biology and Agriculture (CPQBA), University of Campinas (UNICAMP), Campinas, São Paulo, Brazil; Institute of Biology (IB)- University of Campinas (UNICAMP), Campinas, São Paulo, Brazil.

Center for Chemistry, Biology and Agriculture (CPQBA), University of Campinas (UNICAMP), Campinas, São Paulo, Brazil; Institute of Biology (IB)- University of Campinas (UNICAMP), Campinas, São Paulo, Brazil.

出版信息

Ecotoxicol Environ Saf. 2019 Sep 15;179:232-240. doi: 10.1016/j.ecoenv.2019.04.044. Epub 2019 Apr 30.

Abstract

Metagenomics is a powerful approach to study microorganisms present in any given environment and their potential to maintain and improve ecosystem health without the need of cultivating these microorganisms in the laboratory. In this study, we combined a cultivation-independent metagenomics approach with functional assays to identify the detoxification potential of microbial genes evaluating their potential to contribute to xenobiotics resistance in oil-impacted mangrove sediments. A metagenomic fosmid library containing 12,960 clones from highly contaminated mangrove sediment was used in this study. For assessment of metal resistance, clones were grown in culture medium with increasing concentrations of mercury. The analyses metagenomic library sequences revealed the presence of genes related to heavy metals and antibiotics resistance in the oil-impacted mangrove microbiome. The taxonomic profiling of these sequences suggests that at the genus level, Geobacter was the most abundant genus in our dataset. A functional screening assessment of the metagenomic library successfully detected 24 potential heavy metal tolerant clones, six of which were capable of growing with increased concentrations of mercury. The genetic characterization of selected clones allowed the detection of genes related to detoxification processes, such as chromate transport protein ChrA, haloacid dehalogenase-like hydrolase, lipopolysaccharide transport system, and 3-oxoacyl-[acyl-carrier-protein] reductase. Clones were capable of growing in medium containing increased concentrations of metals and antibiotics, but none manifested strong mercury removal from culture medium characteristic of mercuric reductase activity. These results suggest that resistance to xenobiotic stress varies greatly and that additional studies to elucidate the potential of metal biotransformation need to be carried out with the goal of improving bioremediation application.

摘要

宏基因组学是一种强大的方法,可以研究任何给定环境中存在的微生物及其维持和改善生态系统健康的潜力,而无需在实验室中培养这些微生物。在这项研究中,我们将非培养的宏基因组学方法与功能测定相结合,以确定微生物基因的解毒潜力,评估它们对受石油影响的红树林沉积物中异生物抗性的贡献潜力。本研究使用了一个包含 12960 个克隆的高度污染红树林沉积物的宏基因组 fosmid 文库。为了评估金属抗性,将克隆在含有汞浓度逐渐增加的培养基中培养。对宏基因组文库序列的分析揭示了受石油影响的红树林微生物组中存在与重金属和抗生素抗性相关的基因。这些序列的分类分析表明,在属水平上,Geobacter 是我们数据集最丰富的属。对宏基因组文库的功能筛选评估成功检测到 24 个潜在的重金属耐受克隆,其中 6 个能够在汞浓度增加的情况下生长。对选定克隆的遗传特征分析允许检测到与解毒过程相关的基因,如铬酸盐转运蛋白 ChrA、卤代酸脱卤酶样水解酶、脂多糖转运系统和 3-氧酰基-[酰基载体蛋白]还原酶。克隆能够在含有增加浓度的金属和抗生素的培养基中生长,但没有一个表现出从培养基中去除汞的特征,即具有汞还原酶活性。这些结果表明,对外源生物胁迫的抗性差异很大,需要进一步研究以阐明金属生物转化的潜力,目标是改善生物修复应用。

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