: One promising strategy for the in situ bioremediation of radioactive contaminants is to stimulate the activity of dissimilatory metal-reducing microorganisms to reductively precipitate uranium and other soluble toxic metals. While Geobacter bacteria are recognized as important agents for the reductive precipitation of soluble U(VI) to insoluble U(IV) in situ, the underlying mechanism is poorly understood. The reduction of U(VI) and other soluble contaminants by these microorganisms is directly dependent on the reduction of Fe(III) oxides, their natural electron acceptor. The recent discovery that Geobacter bacteria employ a novel mechanism for electron transport to Fe(III) oxides via protein nanowires (conductive pili) prompted us to investigate a potential role for these extracellular appendages in U(VI) reduction. Expression of pilus nanowires in the model organism Geobacter sulfurreducens led to a rapid precipitation of U(VI) to a U(IV) mineral along the nanowire length, demonstrating a previously unrecognized role for Geobacter nanowires in electron transfer to U(VI). Our objective in this proposal is to characterize the mechanism of U(VI) reduction mediated by Geobacter's pilus nanowires at the molecular and nano-scale level. As a complex biological and electronic structure of nanoscale dimensions (3-5 nm in diameter), insights into the structure and electronics of Geobacter's nanowires and their contribution to uranium reduction will require innovative approaches that integrate biological, physical and nanotechnological tools. To accomplish this, we bring together a team of researchers with recognized leadership in microbial metal reduction and nanowires, biosensor design, and environmental spectroscopy. Specifically, we propose the following aims:
Specific Aim #1 : Identify the molecular basis of nanowire-mediated electron transfer. Our working hypothesis, based on strong preliminary data presented in the application, is that the key nanowire components involved in U(VI) reduction can be identified by screening mutants with defects in nanowire functions and studying their effect in U(VI) transformations.
Specific Aim #2 : Develop nanostructured bioelectronic interfaces that integrate electroactive nanowires and lipid membranes with electrodes. Our working hypothesis, also based upon preliminary data, is that nanowire electrochemical processes can be mimicked and studied using nanostructured bioelectronic interfaces that integrate functional components of the nanowire system with electrodes. These studies are innovative in that they integrate microbiological, physical and nanotechnological tools to elucidate the novel mechanism of uranium reduction by microbial nanowires. At the completion of these studies we will have elucidated the basis of nanowire electron transfer to uranium at the molecular and nanoscale level, to enhance our knowledge of the key biological processes involved in the bioremediation of radionuclides and other soluble toxic metals and provide a basis for improved bioremediation processes.

Public Health Relevance

This work will use nanotechnology-enabled tools to develop a fundamental understanding of radionuclide reduction via Geobacter nanowires. These results will enable design of efficient in situ bioremediation strategies for radionuclides, thereby reducing a widespread and serious threat to the Nation's health.

Agency
National Institute of Health (NIH)
Institute
National Institute of Environmental Health Sciences (NIEHS)
Type
Research Project (R01)
Project #
5R01ES017052-02
Application #
7759125
Study Section
Special Emphasis Panel (ZES1-SET-G (R8))
Program Officer
Henry, Heather F
Project Start
2009-02-01
Project End
2011-11-30
Budget Start
2009-12-01
Budget End
2010-11-30
Support Year
2
Fiscal Year
2010
Total Cost
$297,584
Indirect Cost
Name
Michigan State University
Department
Microbiology/Immun/Virology
Type
Schools of Arts and Sciences
DUNS #
193247145
City
East Lansing
State
MI
Country
United States
Zip Code
48824
Speers, Allison M; Schindler, Bryan D; Hwang, Jihwan et al. (2016) Genetic Identification of a PilT Motor in Geobacter sulfurreducens Reveals a Role for Pilus Retraction in Extracellular Electron Transfer. Front Microbiol 7:1578
Cologgi, Dena L; Speers, Allison M; Bullard, Blair A et al. (2014) Enhanced uranium immobilization and reduction by Geobacter sulfurreducens biofilms. Appl Environ Microbiol 80:6638-46
Reguera, Gemma (2012) Electron transfer at the cell-uranium interface in Geobacter spp. Biochem Soc Trans 40:1227-32
Veazey, Joshua P; Reguera, Gemma; Tessmer, Stuart H (2011) Electronic properties of conductive pili of the metal-reducing bacterium Geobacter sulfurreducens probed by scanning tunneling microscopy. Phys Rev E Stat Nonlin Soft Matter Phys 84:060901
Reguera, Gemma (2011) When microbial conversations get physical. Trends Microbiol 19:105-13
Cologgi, Dena L; Lampa-Pastirk, Sanela; Speers, Allison M et al. (2011) Extracellular reduction of uranium via Geobacter conductive pili as a protective cellular mechanism. Proc Natl Acad Sci U S A 108:15248-52