NMR methods for in situ biofilm metabolism studies.
Novel procedures and instrumentation are described for nuclear magnetic resonance (NMR) spectroscopy and imaging studies of live, in situ microbial films. A perfused NMR/optical microscope sample chamber containing a planar biofilm support was integrated into a recirculation/dilution flow loop growth reactor system and used to grow in situ Shewanella oneidensis strain MR-1 biofilms. Localized NMR techniques were developed and used to non-invasively monitor time-resolved metabolite concentrations and to image the biomass volume and distribution. As a first illustration of the feasibility of the methodology an initial 13C-labeled lactate metabolic pathway study was performed, yielding results consistent with existing genomic data for MR-1. These results represent progress toward our ultimate goal of correlating time- and depth-resolved metabolism and mass transport with gene expression in live in situ biofilms using combined NMR/optical microscopy techniques.
Majors
Paul D
PD
Pacific Northwest National Laboratory, MSIN: K8-98, Richland, WA 99352, USA.
McLean
Jeffrey S
JS
Pinchuk
Grigoriy E
GE
Fredrickson
James K
JK
Gorby
Yuri A
YA
Minard
Kevin R
KR
Wind
Robert A
RA
D018441:Biofilms / Q000254:growth & development; D019149:Bioreactors; D002247:Carbon Isotopes; D003470:Culture Media; D005650:Fumarates / Q000378:metabolism; D019344:Lactic Acid / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy / Q000295:instrumentation / Q000379:methods*; D008828:Microbiological Techniques / Q000295:instrumentation; D020592:Shewanella / Q000378:metabolism
D002247:Carbon Isotopes; D003470:Culture Media; D005650:Fumarates; D019344:Lactic Acid; C032005:fumaric acid