Henriques Ana C, De Marco Paolo
Instituto de Investigação e Formação Avançada em Ciências e Tecnologias da Saúde, CESPU, Rua Central de Gandra 1317, 4585-116 Paredes, Portugal.
PLoS One. 2015 May 15;10(5):e0125735. doi: 10.1371/journal.pone.0125735. eCollection 2015.
Quantitatively, methanesulfonate (MSA) is a very relevant compound in the global biogeochemical sulfur cycle. Its utilization by bacteria as a source of carbon and energy has been described and a specific enzyme, methanesulfonate monooxygenase (MSAMO), has been found to perform the first catabolic step of its oxidation. Other proteins seemingly involved in the import of MSA into bacterial cells have been reported. In this study, we obtained novel sequences of genes msmA and msmE from marine, estuary and soil MSA-degraders (encoding the large subunit of the MSAMO enzyme and the periplasmic component of the import system, respectively). We also obtained whole-genome sequences of two novel marine Filomicrobium strains, Y and W, and annotated two full msm operons in these genomes. Furthermore, msmA and msmE sequences were amplified from North Atlantic seawater and analyzed. Good conservation of the MsmA deduced protein sequence was observed in both cultured strains and metagenomic clones. A long spacer sequence in the Rieske-type [2Fe-2S] cluster-binding motif within MsmA was found to be conserved in all instances, supporting the hypothesis that this feature is specific to the large (α) subunit of the MSAMO enzyme. The msmE gene was more difficult to amplify, from both cultivated isolates and marine metagenomic DNA. However, 3 novel msmE sequences were obtained from isolated strains and one directly from seawater. With both genes, our results combined with previous metagenomic analyses seem to imply that moderate to high-GC strains are somehow favored during enrichment and isolation of MSA-utilizing bacteria, while the majority of msm genes obtained by cultivation-independent methods have low levels of GC%, which is a clear example of the misrepresentation of natural populations that culturing, more often than not, entails. Nevertheless, the data obtained in this work show that MSA-degrading bacteria are abundant in surface seawater, which suggests ecological relevance for this metabolic group of bacteria.
从数量上看,甲磺酸盐(MSA)是全球生物地球化学硫循环中一种非常重要的化合物。细菌将其作为碳源和能源利用的情况已有描述,并且已发现一种特定的酶,即甲磺酸盐单加氧酶(MSAMO),可执行其氧化的第一步分解代谢。还报道了其他似乎参与MSA导入细菌细胞的蛋白质。在本研究中,我们从海洋、河口和土壤中的MSA降解菌中获得了msmA和msmE基因的新序列(分别编码MSAMO酶的大亚基和导入系统的周质成分)。我们还获得了两种新型海洋丝状菌菌株Y和W的全基因组序列,并在这些基因组中注释了两个完整的msm操纵子。此外,从北大西洋海水中扩增并分析了msmA和msmE序列。在培养菌株和宏基因组克隆中均观察到推导的MsmA蛋白序列具有良好的保守性。发现在所有情况下,MsmA中 Rieske 型[2Fe-2S]簇结合基序中的一个长间隔序列都是保守的,这支持了该特征是MSAMO酶大亚基(α)特有的这一假设。从培养的分离株和海洋宏基因组DNA中扩增msmE基因都更困难。然而,从分离菌株中获得了3个新的msmE序列,其中一个直接从海水中获得。对于这两个基因,我们的结果与先前的宏基因组分析相结合似乎意味着,在富集和分离利用MSA的细菌过程中,中等至高GC含量的菌株在某种程度上更受青睐,而通过非培养方法获得的大多数msm基因的GC%水平较低,这是培养往往会导致自然种群代表性失真的一个明显例子。尽管如此,这项工作获得的数据表明,MSA降解细菌在表层海水中大量存在,这表明该代谢菌群具有生态相关性。