Project description:The thermophilic Aquificales inhabit and play important biogeochemical roles in the geothermal environments globally. Although intensive studies on physiology, microbial ecology, biochemistry, metagenomics and metatranscriptomics of the Aquificales¬ species and Aquificales-containing environmental samples have been conducted, comprehensive understandings about their ecophysiology, especially in the natural niches have been limited. In the present study, an integrated suite of metagenomic, metatranscriptomic and metaproteomic analyses, for the first time, were conducted on a filamentous microbial community from the Apron and Channel Facies (ACF) of CaCO3 (travertine) deposition at Narrow Gauge, Mammoth Hot Springs, Yellowstone National Park.
Project description:The thermophilic Aquificales inhabit and play important biogeochemical roles in the geothermal environments globally. Although intensive studies on physiology, microbial ecology, biochemistry, metagenomics and metatranscriptomics of the Aquificales¬ species and Aquificales-containing environmental samples have been conducted, comprehensive understandings about their ecophysiology, especially in the natural niches have been limited. In the present study, an integrated suite of metagenomic, metatranscriptomic and metaproteomic analyses, for the first time, were conducted on a filamentous microbial community from the Apron and Channel Facies (ACF) of CaCO3 (travertine) deposition at Narrow Gauge, Mammoth Hot Springs, Yellowstone National Park.
Project description:This experiment aims on the identification of serine hydrolases from a complex thermophile community that live in a hot vent in Kamchatka Peninsula based on in vivo labelling with FP-alkyne directly in the hot spring and subsequent analysis using metagenomics/metaproteomics. To this end, sediment samples were collected and treated using the following three conditions. DMSO- treated control FP-alkyne labelled Samples for each condition were prepared in triplicate, resulting a total number of 6 samples per spring. Labelling was performed using 4 µM of the probe FP-alkyne and incubation for 2 h in the hot spring.
Project description:Thermophilic cyanobacteria are not only fundamental models for investigating thermotolerance mechanisms but also a valuable reservoir for the development of novel thermophilic bioresources. Progress in both of these areas, however, is contingent upon the precise characterization of their key subcellular organelles, such as the intracellular polyphosphate-rich granules which we term stabilisomes. The lack of a high-purity, structure-preserving isolation method has been a major technical bottleneck hindering such research. This study describes a high-fidelity, structure-preserving purification strategy, boosting the granule-to-protein yield by over 10,000-fold compared with conventional methods. The high fidelity of this method is confirmed by the retention of complex proteomic (937 proteins) and metabolomic (1,076 metabolites) signatures. Building on this, subsequent quantitative analysis across cyanobacteria at 7 hot spring sampling sites revealed a conserved core chemical composition dominated by polyphosphate (21.1%-36.4%), proteins (9.9%-20.0%), amino acids (1.6%-7.4%), and lipids (12.0%-21.0%), which nevertheless showed significant variabilitiy in the abundance of these components across species. This work establishes a robust methodological framework for isolating prokaryotic stress-responsive organelles, lays the foundation for mechanistic investigations into microbial adaptation, and serves as a prerequisite for exploring the biomedical applications of these organelles.