Project description:Carbon monoxide (CO) oxidising microorganisms are present in volcanic deposits throughout succession, with levels of vegetation and soil influencing the communities present. Carboxydovores are a subset of CO oxidisers that use CO as an energy source, which raises questions about the physiological and metabolic features that make them more competitive in harsh volcanic ecosystems. To address these questions, samples were taken from volcanic strata formed by eruptions from Calbuco Volcano (Chile) in 2015 (tephra) and 1917 (soil). Two carboxydovore members of the Burkholderiaceae family were isolated for further study to elucidate the benefits of carboxydovory for the survival of these strains in extreme volcanic ecosystems. The isolates were identified as Paraburkholderia terrae COX (isolated from the 2015 tephra) and Cupriavidus str. CV2 (isolated from the 1917 soil). 16S rRNA gene sequencing showed that within the family Burkholderiacea, the genus Paraburkholderia dominated the 2015 volcanic deposit with an average relative abundance of 73.81%, whereas in the 1917 volcanic deposit, Cupriavidus accounted for 33.64% (average relative abundance). Both strains oxidise CO across a broad range of concentrations (< 100 ppmv - 10,000 ppmv), and genome sequence analysis revealed a candidate form-I carbon monoxide dehydrogenase (CODH), which is likely to catalyse this process. Each strain oxidised CO specifically at stationary phase but the conditions for induction of CODH expression were distinct. Cupriavidus strain CV2 expressed CODH only when CO was added to cultures (100 ppm), while Pb. terrae COX expressed CODH regardless of supplementary CO addition. Based on comparative metabolic and phylogenetic analyses, Cupriavidus strain CV2 is proposed as a novel species within the genus Cupriavidus with the name Cupriavidus ulmosensis sp. nov. for the type strain CV2T (= NCIMB 15506 T, = CECT 30956 T). This study provides valuable insights into the physiology and metabolism of carboxydovores which colonise volcanic ecosystems.

Project description:Diaminopimelic acid (DAP) is a unique component of the cell wall of Gram-negative bacteria. It is also an important component of organic matter and is widely utilized by microbes in the world's oceans. However, neither DAP concentrations nor marine DAP-utilizing microbes have been investigated. Here, DAP concentrations in seawater were measured and the diversity of marine DAP-utilizing bacteria and the mechanisms for their DAP metabolism were investigated. Free DAP concentrations in seawater, from surface to a 5,000 m depth, were found to be between 0.61 μM and 0.96 μM in the western Pacific Ocean. DAP-utilizing bacteria from 20 families in 4 phyla were recovered from the western Pacific seawater and 14 strains were further isolated, in which Pseudomonadota bacteria were dominant. Based on genomic and transcriptomic analyses combined with gene deletion and in vitro activity detection, DAP decarboxylase (LysA), which catalyzes the decarboxylation of DAP to form lysine, was found to be a key and specific enzyme involved in DAP metabolism in the isolated Pseudomonadota strains. Interrogation of the Tara Oceans database found that most LysA-like sequences (92%) are from Pseudomonadota, which are widely distributed in multiple habitats. This study provides an insight into DAP metabolism by marine bacteria in the ocean and contributes to our understanding of the mineralization and recycling of DAP by marine bacteria. IMPORTANCE DAP is a unique component of peptidoglycan in Gram-negative bacterial cell walls. Due to the large number of marine Gram-negative bacteria, DAP is an important component of marine organic matter. However, it remains unclear how DAP is metabolized by marine microbes. This study investigated marine DAP-utilizing bacteria by cultivation and bioinformational analysis and examined the mechanism of DAP metabolism used by marine bacteria. The results demonstrate that Pseudomonadota bacteria are likely to be an important DAP-utilizing group in the ocean and that DAP decarboxylase is a key enzyme involved in DAP metabolism. This study also sheds light on the mineralization and recycling of DAP driven by bacteria.

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Pseudomonadota

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