Progress toward clonable inorganic nanoparticles.
Pseudomonas moraviensis stanleyae was recently isolated from the roots of the selenium (Se) hyperaccumulator plant Stanleya pinnata. This bacterium tolerates normally lethal concentrations of SeO3(2-) in liquid culture, where it also produces Se nanoparticles. Structure and cellular ultrastructure of the Se nanoparticles as determined by cellular electron tomography shows the nanoparticles as intracellular, of narrow dispersity, symmetrically irregular and without any observable membrane or structured protein shell. Protein mass spectrometry of a fractionated soluble cytosolic material with selenite reducing capability identified nitrite reductase and glutathione reductase homologues as NADPH dependent candidate enzymes for the reduction of selenite to zerovalent Se nanoparticles. In vitro experiments with commercially sourced glutathione reductase revealed that the enzyme can reduce SeO3(2-) (selenite) to Se nanoparticles in an NADPH-dependent process. The disappearance of the enzyme as determined by protein assay during nanoparticle formation suggests that glutathione reductase is associated with or possibly entombed in the nanoparticles whose formation it catalyzes. Chemically dissolving the nanoparticles releases the enzyme. The size of the nanoparticles varies with SeO3(2-) concentration, varying in size form 5 nm diameter when formed at 1.0 μM [SeO3(2-)] to 50 nm maximum diameter when formed at 100 μM [SeO3(2-)]. In aggregate, we suggest that glutathione reductase possesses the key attributes of a clonable nanoparticle system: ion reduction, nanoparticle retention and size control of the nanoparticle at the enzyme site.
Department of Chemistry, Colorado State University, Fort Collins, CO 80523, USA. ackerson@colostate.edu.
Thomas W
TW
Ni
Lucian C
LC
Staicu
Richard S
RS
Nemeth
Cindi L
CL
Schwartz
David
D
Crawford
Jeffrey D
JD
Seligman
William J
WJ
Hunter
Elizabeth A H
EA
Pilon-Smits
Christopher J
CJ
Ackerson
D053758:Nanoparticles / Q000737:chemistry*; D010316:Particle Size; D011549:Pseudomonas / Q000378:metabolism*; D020887:Selenious Acid / Q000378:metabolism*; D012643:Selenium / Q000737:chemistry* / Q000378:metabolism
D020887:Selenious Acid; D012643:Selenium