High-resolution imaging of Rhodobacter sphaeroides strains with increased lipid accumulation
EMSL Project ID
48544
Abstract
We are requesting access to quiet wing microscopy, including ETEM and STEM electron microscopes and the helium ion microscope, as well as the super resolution structured illumination fluorescence and multi-photon/FLIM integrated confocal microscopes at EMSL. We propose to utilize these facilities, in collaboration with EMSL experts, to obtain molecular and high-resolution insights into overproduction of potential advanced biofuel precursors in bacteria. Specifically, we will visualize the location and organization of fatty acids in “high lipid” producing mutants that we have isolated in the photosynthetic bacterium Rhodobacter sphaeroides. We are using this model since R. sphaeroides has the somewhat usually ability to naturally increase fatty acid content under anaerobic conditions. Access to the state of the art imaging facilities at EMSL is crucial to unraveling the biological processes leading to lipid accumulation in these strains. Our strategy is to use the insight we gain of fatty acid accumulation in R. sphaeroides to develop approaches to improve the yield of fatty acids or other fuel precursors in this or possibly other microbes being studied in GLBRC. This project is supported by the Office of Biological and Environmental Research through its funding of Great Lakes Bioenergy Research Center (GLBRC) with the goal of generating knowledge needed to produce next generation biofuels or fuel precursors. EMSL supports this mission by providing innovative solutions to energy production. The proposed collaborative project leverages genome-enabled and system biology activities within GLBRC and the innovative imaging capabilities of EMSL to provide knowledge about how biofuel-producing bacteria store lipids. Preliminary data generated in a pilot project demonstrate that there are striking differences in the ultrastructure and organization of lipids in some of our mutants that we are able to visualize using the unique capabilities at EMSL. Access to the EMSL state of the art microscopy facilities is crucial to the success of these studies, as GLBRC has neither the high-resolution facilities nor the expertise to conduct these studies. Use of the EMSL microscopy imaging facilities will allow us to gain critical insight into the cellular state of the increased fatty acid content in our "high lipid" mutants of R. sphaeroides. This knowledge will be integral to ongoing studies that seek to elucidate the genetic and biochemical mechanisms that regulate the flow of strategic precursors into fatty acids in bacteria. These studies have potential applications in increasing yield of advanced biofuels in algae as well as other photosynthetic or non-photosynthetic microbes that are targeted for investigation by DOE.
Project Details
Project type
Special Science
Start Date
2014-07-24
End Date
2015-09-30
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members
Related Publications
Lemmer KC, AC Dohnalkova, DR Noguera, and TJ Donohue. 2015. "Oxygen-dependent regulation of bacterial lipid production." Journal of Bacteriology:JB.02510-02514. Accepted 2 March 2015. doi:10.1128/jb.02510-14