New monitoring approach for metabolic dynamics in microbial ecosystems using stable-isotope-labeling technologies.
We have developed a new approach for monitoring the metabolic dynamics in microbial ecosystems using a combination of DNA fingerprinting and metabolome analysis based on stable-isotope-labeling technologies. Stable-isotope probing of DNA (DNA-SIP) has been used previously for the evaluation of cross-feeding in microbial communities. For the development and validation of our monitoring approach, fecal microbiota were analyzed with stable-isotope-labeled glucose used as the sole carbon source. In order to link the metabolic information and the microbial variability, we performed metabolic-microbial correlation analysis based on nuclear magnetic resonance (NMR) profiles and denaturing gradient gel electrophoresis (DGGE) fingerprints, which successfully identified the glucose-utilizing bacteria and their related extracellular metabolites. Moreover, our approach revealed information regarding the carbon flux, in that the "first" wave of extracellular metabolites secreted by the glucose-utilizing bacteria were incorporated into the "secondary" group of substrate-utilizing bacteria, and that this "secondary" group further produced their own secondary metabolized substrates. Thus, this approach is a powerful tool for monitoring the metabolic dynamics in microbial ecosystems and allows for the tracking of the carbon flux within a microbial community.
Date
Yasuhiro
Y
Department of Life Science and Medical Bioscience, Waseda University, 2-2 Wakamatsu-cho, Shinjuku-ku, Tokyo, 162-8480, Japan.
Nakanishi
Yumiko
Y
Fukuda
Shinji
S
Kato
Tamotsu
T
Tsuneda
Satoshi
S
Ohno
Hiroshi
H
Kikuchi
Jun
J
D000818:Animals; D001419:Bacteria / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D002244:Carbon / Q000378:metabolism; D002247:Carbon Isotopes / Q000378:metabolism; D016172:DNA Fingerprinting; D017753:Ecosystem; D005947:Glucose / Q000378:metabolism; D007553:Isotope Labeling / Q000379:methods*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010802:Phylogeny; D013997:Time Factors
D002247:Carbon Isotopes; D002244:Carbon; D005947:Glucose