Maixner Frank, Wagner Michael, Lücker Sebastian, Pelletier Eric, Schmitz-Esser Stephan, Hace Karin, Spieck Eva, Konrat Robert, Le Paslier Denis, Daims Holger
Department für Mikrobielle Okologie, Universität Wien, Vienna, Austria.
Environ Microbiol. 2008 Nov;10(11):3043-56. doi: 10.1111/j.1462-2920.2008.01646.x. Epub 2008 May 5.
Nitrite-oxidizing bacteria of the genus Nitrospira are ubiquitous in natural ecosystems and also in wastewater treatment plants. Nitrospira are members of a distinct phylum, not closely related to other nitrifiers, and no genomic sequences from this genus have been available so far. Here we applied an environmental genomics approach to sequence and assemble a 137 kbp-long genome fragment of 'Candidatus Nitrospira defluvii', which had been enriched from activated sludge and belongs to Nitrospira sublineage I without isolated representatives. The annotation of this contig, which carried the 16S rRNA gene of N. defluvii, offered first insight into the genome of Nitrospira. Surprisingly, we found a gene similar to genes encoding chlorite dismutase (CLD), an enzyme degrading chlorite (ClO(2)(-)) to Cl(-) and O(2). To date, CLDs with high catalytic activity have been found only in perchlorate- and chlorate-reducing bacteria but not in nitrifiers. Heterologous expression in E. coli followed by enzymatic tests confirmed that this gene of Nitrospira encodes a highly active CLD, which is also expressed in situ by Nitrospira, indicating that this nitrite oxidizer might be involved in the bioremediation of perchlorate and chlorite. Phylogenetic analyses showed that CLD and related proteins are widely distributed among the Bacteria and Archaea, and indicated that this enzyme family appeared relatively early in evolution, has been subject to functional diversification and might play yet unknown roles in microbial metabolism.
硝化螺菌属的亚硝酸盐氧化细菌在自然生态系统以及污水处理厂中普遍存在。硝化螺菌属于一个独特的门,与其他硝化菌的关系并不密切,目前尚无该属的基因组序列。在此,我们应用环境基因组学方法对“假硝化螺菌 defluvii”的一个137 kbp长的基因组片段进行测序和组装,该菌株从活性污泥中富集得到,属于硝化螺菌亚系I,尚无分离的代表菌株。该重叠群携带了 N. defluvii 的 16S rRNA 基因,对其注释首次揭示了硝化螺菌的基因组情况。令人惊讶的是,我们发现了一个与编码亚氯酸盐歧化酶(CLD)的基因相似的基因,该酶可将亚氯酸盐(ClO₂⁻)降解为 Cl⁻ 和 O₂。迄今为止,仅在高氯酸盐和氯酸盐还原细菌中发现了具有高催化活性的 CLD,而在硝化菌中未发现。在大肠杆菌中进行异源表达并随后进行酶学测试证实,硝化螺菌的这个基因编码一种高活性的 CLD,并且硝化螺菌在原位也表达该基因,这表明这种亚硝酸盐氧化菌可能参与高氯酸盐和亚氯酸盐的生物修复。系统发育分析表明,CLD 及相关蛋白在细菌和古菌中广泛分布,并表明该酶家族在进化过程中出现相对较早,已经经历了功能多样化,可能在微生物代谢中发挥尚未明确的作用。