Co-localization of particulate methane monooxygenase and cd1 nitrite reductase in the denitrifying methanotroph 'Candidatus Methylomirabilis oxyfera'.
Co-localization of particulate methane monooxygenase and cd1 nitrite reductase in the denitrifying methanotroph 'Candidatus Methylomirabilis oxyfera'.
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DOI：
10.1111/j.1574-6968.2012.02615.x
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发表时间：
2012-09
影响因子：
2.1
通讯作者：
Ming L. Wu;T. V. van Alen;E. V. van Donselaar;M. Strous;M. Jetten;L. van Niftrik
中科院分区：
文献类型：
--
作者：
Ming L. Wu;T. V. van Alen;E. V. van Donselaar;M. Strous;M. Jetten;L. van Niftrik
“Methylomirabilis oxyfera”是一种具有独特能力的细菌，通过新发现的好氧内途径将厌氧甲烷氧化与反硝化偶联。最近，通过宏基因组测序将Methylomirabilis oxyfera的全基因组组装成一个2.7-Mb的环状单染色体。分枝杆菌基因组Oxyfera揭示了进行甲烷氧化和通过一氧化氮将亚硝酸盐转化为氧气和二氮气体的全部潜力。在这项研究中，我们通过免疫金定位表明，甲烷和亚硝酸盐转化途径的关键酶确实存在于单个M。尖细胞针对颗粒甲烷单加氧酶（pMMO）和cd（1）亚硝酸盐还原酶（NirS）的抗血清被提出，并用于单标记和双标记实验中的免疫金定位。我们以前的研究表明，M。oxifera不像在经典的变形菌甲烷氧化菌中观察到的那样形成含有pMMO的胞质膜。我们的结果表明，在M.在oxyfera中，pMMO和NirS酶分别定位于细胞质膜和周质。此外，双标记显示pMMO和NirS在单个M中共同出现。尖细胞
'Candidatus Methylomirabilis oxyfera'; is a polygon-shaped bacterium that was shown to have the unique ability to couple anaerobic methane oxidation to denitrification, through a newly discovered intra-aerobic pathway. Recently, the complete genome of Methylomirabilis oxyfera was assembled into a 2.7-Mb circular single chromosome by metagenomic sequencing. The genome of M. oxyfera revealed the full potential to perform both methane oxidation and the conversion of nitrite via nitric oxide into oxygen and dinitrogen gas. In this study, we show by immunogold localization that key enzymes from both methane- and nitrite-converting pathways are indeed present in single M. oxyfera cells. Antisera targeting the particulate methane monooxygenase (pMMO) and the cd(1) nitrite reductase (NirS) were raised and used for immunogold localization in both single- and double-labelling experiments. Our previous studies have shown that M. oxyfera does not develop pMMO-containing intracytoplasmic membranes as is observed in classical proteobacterial methanotrophs. Our results suggest that in M. oxyfera, the pMMO and NirS enzymes localized to the cytoplasmic membrane and periplasm, respectively. Further, double-labelling showed co-occurrence of pMMO and NirS in single M. oxyfera cells.