Tracing incorporation of heavy water into proteins for species-specific metabolic activity in complex communities

0532445 - MBÚ 2021 RIV NL eng J - Článek v odborném periodiku
Starke, Robert - Oliphant, K. - Jehmlich, N. - Schaepe, S. S. - Sachsenberg, T. - Kohlbacher, O. - Allen-Vercoe, E. - von Bergen, M.
Tracing incorporation of heavy water into proteins for species-specific metabolic activity in complex communities.
Journal of Proteomics. Roč. 222, JUN 30 (2020), č. článku 103791. ISSN 1874-3919. E-ISSN 1876-7737
Grant CEP: GA ČR(CZ) GJ20-02022Y
Institucionální podpora: RVO:61388971
Klíčová slova: gut microbiota * mass-spectrometry * sip * degradation * nitrogen
Obor OECD: Microbiology
Impakt faktor: 4.044, rok: 2020
Způsob publikování: Omezený přístup
https://www.sciencedirect.com/science/article/pii/S1874391920301597

Stable isotope probing (SIP) approaches are a suitable tool to identify active organisms in bacterial communities, but adding isotopically labeled substrate can alter both the structure and the functionality of the community. Here, we validated and demonstrated a substrate-independent protein-SIP protocol using isotopically labeled water that captures the entire microbial activity of a community. We found that O-18 yielded a higher incorporation rate into peptides and thus comprised a higher sensitivity. We then applied the method to an in vitro model of a human distal gut microbial ecosystem grown in two medium formulations, to evaluate changes in microbial activity between a high-fiber and high-protein diet. We showed that only little changes are seen in the community structure but the functionality varied between the diets. In conclusion, our approach can detect species-specific metabolic activity in complex bacterial communities and more specifically to quantify the amount of amino acid synthesis. Heavy water makes possible to analyze the activity of bacterial communities for which adding an isotopically labeled energy and nutrient sources is not easily feasible.
Trvalý link: http://hdl.handle.net/11104/0310934