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Chemolithotrophs are microorganisms that obtain energy by oxidizing inorganic molecules such as hydrogen gas (H₂), ammonia (NH₃), reduced sulfur compounds (H₂S, S²⁻), and ferrous iron (Fe²⁺). Unlike heterotrophic organisms that rely on organic carbon, chemolithotrophs transfer electrons from these inorganic donors to the electron transport chain (ETC), generating a proton motive force (PMF) that drives ATP synthesis through oxidative phosphorylation.
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Nitrogen is an essential element in biological systems, forming a crucial component of proteins, nucleic acids, and other cellular constituents. Many bacteria and archaea acquire nitrogen in the form of nitrate (NO₃⁻) or ammonia (NH₃), which are then assimilated into biomolecules through specific enzymatic pathways.Assimilatory Nitrate ReductionWhen nitrate enters the cell, it undergoes a two-step reduction process known as assimilatory nitrate reduction. Initially, the enzyme...
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Microorganisms rely on proteins as an essential carbon and energy source, particularly in environments with limited polysaccharides or lipids. However, proteins are too large to cross the plasma membrane unaided, necessitating enzymatic degradation. Microbes secrete extracellular proteases and peptidases that hydrolyze proteins into peptides, which can then be transported across the membrane. Once inside the cell, intracellular proteases degrade these peptides into free amino acids, which...
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Anoxygenic phototrophic bacteria are a diverse group of microorganisms that perform photosynthesis without producing oxygen. They primarily include purple sulfur bacteria, purple nonsulfur bacteria, green sulfur bacteria, and green nonsulfur bacteria. These bacteria are classified into the Gammaproteobacteria, Alphaproteobacteria, Betaproteobacteria, Chlorobi, and Chloroflexi lineages, each with distinct physiological and ecological adaptations.Purple sulfur bacteria belong to the...
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Organisms exhibit remarkable metabolic diversity, categorized based on how they acquire energy and carbon. These strategies enable survival in various ecological niches and are essential for maintaining energy flow and nutrient cycling within ecosystems.Energy and Carbon SourcesOrganisms are classified as phototrophs or chemotrophs based on energy acquisition. Phototrophs use light as their energy source, while chemotrophs rely on oxidizing chemical compounds. Further differentiation arises...
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Universal activity-based labeling method for ammonia- and alkane-oxidizing bacteria.

Dimitra Sakoula1,2, Garrett J Smith3, Jeroen Frank3,4

  • 1Department of Microbiology, RIBES, Radboud University, Heyendaalseweg 135, 6525 AJ, Nijmegen, the Netherlands. dimitra.sakoula@univie.ac.at.

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This summary is machine-generated.

A new method uses a chemical probe to tag active ammonia- and alkane-oxidizing bacteria in environmental samples. This technique allows for cultivation-independent detection, identification, and enrichment of these important microbes.

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Area of Science:

  • Microbiology
  • Biogeochemistry
  • Molecular Biology

Background:

  • Microbial roles in nitrogen and carbon cycles are incompletely understood.
  • Detecting low-abundance ammonia- and alkane-oxidizing bacteria in situ is challenging.

Purpose of the Study:

  • To develop a cultivation-independent method for detecting and identifying ammonia- and alkane-oxidizing bacteria.
  • To enable enrichment of these bacteria from complex environmental samples.

Main Methods:

  • Activity-based protein profiling using 1,7-octadiyne as a bifunctional enzyme probe.
  • Alkyne-azide cycloaddition for fluorescent or biotin labeling of active monooxygenases.
  • Combined techniques including immunogold labeling, fluorescence in situ hybridization, and fluorescence-activated cell sorting.

Main Results:

  • Successful labeling of active ammonia and alkane monooxygenases.
  • Subcellular localization of tagged enzymes confirmed in model strains.
  • Direct link established between functional lifestyles and phylogenetic identification.
  • Enrichment of nitrifiers and alkane-oxidizing bacteria from environmental samples, enabling metagenome-assembled genome recovery.

Conclusions:

  • A novel functional tagging technique reliably detects, identifies, and enriches ammonia- and alkane-oxidizing bacteria.
  • This method advances the study of microbial biodiversity in biogeochemical cycles.
  • The protocol facilitates the recovery of high-quality metagenome-assembled genomes from unculturable bacteria.