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

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388971%3A_____%2F20%3A00532445" target="_blank" >RIV/61388971:_____/20:00532445 - isvavai.cz</a>

Výsledek na webu

<a href="https://www.sciencedirect.com/science/article/pii/S1874391920301597" target="_blank" >https://www.sciencedirect.com/science/article/pii/S1874391920301597</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1016/j.jprot.2020.103791" target="_blank" >10.1016/j.jprot.2020.103791</a>

Jazyk výsledku

angličtina

Název v původním jazyce

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

Popis výsledku v původním jazyce

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.

Název v anglickém jazyce

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

Popis výsledku anglicky

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.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10606 - Microbiology

Projekt

<a href="/cs/project/GJ20-02022Y" target="_blank" >GJ20-02022Y: Úsvit mrtvých: Chemické složení a obrat mrtvé mikrobiální biomasy a její role v potravním řetězci v půdě</a><br>

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2020

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Journal of Proteomics

ISSN

1874-3919

e-ISSN

—

Svazek periodika

222

Číslo periodika v rámci svazku

JUN 30

Stát vydavatele periodika

NL - Nizozemsko

Počet stran výsledku

10

Strana od-do

103791

Kód UT WoS článku

000538943100015

EID výsledku v databázi Scopus

2-s2.0-85083879609