Interrogation of Glucose Metabolism by Oral Biofilms Using Combined NMR/Optical Spectroscopy and Stable Isotope Labeling
EMSL Project ID
16090c
Abstract
The overall project goal is to develop a robust technology capable of characterizing overall community metabolic function and the roles of individual species in mixed-community biofilms. This effort is expected to benefit biofilm research in particular and the discipline of functional community analysis in general. PNNL-developed nuclear magnetic resonance (NMR) and emerging secondary isotope probing (SIP) techniques will be applied in combination to study metabolic function in live, multiple-species oral biofilms. In situ, non-invasive 1H NMR will provide time- and depth-resolved metabolite profiles and metabolic pathway information for 13C-labeled substrates (via 1H-detected-13C NMR). SIP will identify the active biofilm subspecies via the incorporation and subsequent detection of 13C label (from substrate) into their mRNA. Thus, synergistic NMR and SIP methods will provide insight into oral biofilm physiology and the contributing species. The specific scientific aim is to characterize organic acid production on enamel surfaces by:
1. Streptococcus mutans biofilms to address fundamental questions about what types of acids are produced in biofilms over time in response to different feeding regimens
2. Dual-species biofilms also containing Lactobacillus plantarum to evaluate the synergistic and antagonistic effects of this strain on acid composition and residence time on enamel surfaces
3. Multi-species biofilms comprised of a 10-member defined biofilm and/or potentially an undefined oral biofilm to further explore acid formation in conditions that better simulate natural oral biofilms.
As the majority of bacteria in natural ecosystems exist in complex assemblages consisting of more than one organism, knowledge gained from developing approaches to characterize oral biofilms can also be applied to DOE mission areas such as those focused on developing a fundamental understanding of microbial systems (the Genomics: GTL program) and applications of this knowledge to developing methods clean up the environment (the Natural and Accelerated Bioremediation Research (NABIR) program). Further, the non-invasive and non-contacting nature of NMR is ideal for the modeling of communities in hostile environments, such as acid mine drainage (AMD) communities.
Project Details
Start Date
2007-10-01
End Date
2008-11-17
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members
Related Publications
Faculty of 1000 Biology: evaluations for McLean JS et al 2008 Feb 2 (2) :121-31
http://www.f1000biology.com/article/id/1104258/evaluation
Faculty of 1000 Biology: evaluations for McLean JS et al ISME J 2008 Feb 2 (2) :121-31 http://www.f1000biology.com/article/id/1104258/evaluation
McLean JS, ON Ona and PD Majors. 2008. "Correlated biofilm imaging, transport and metabolism measurements via combined nuclear magnetic resonance and confocal microscopy." ISME Journal 2(2), 121-131, doi:10.1038/ismej.2007.107.
McLean JS, PD Majors, CL Reardon, CL Bilksis, SB Reed, MF Romine and JK Fredrickson (2008). “Investigations of Metabolism and Structure within Shewanella oneidensis MR-1 Biofilms,” Journal of Microbiological Methods 74(1): 47-56, doi:10.1016/j.mimet.2008.02.015 .