Towards nanoscale biomedical devices in medicine: biofunctional and spectroscopic characterization of superparamagnetic nanoparticles.
Medical interest in nanotechnology originates from a belief that nanoscale therapeutic devices can be constructed and directed towards its target inside the human body. Such nanodevices can be engineered by coupling superparamagnetic nanoparticle to biomedically active proteins. We hereby report the immobilization of a PhEst, a S-formylglutathione hydrolase from the psychrophilic P. haloplanktis TAC125 onto the gold coated surface of modified superparamagnetic core-shell nanoparticles (Fe(3)O(4)@Au). The synthesis of the nanoparticles is also reported. S-formylglutathione hydrolases constitute a family of ubiquitous enzymes which play a key role in formaldehyde detoxification both in prokaryotes and eukaryotes. PhEst was originally annotated as a putative feruloyl esterase, an enzyme that releases ferulic acid (an antioxidant reactive towards free radicals such as reactive oxygen species) from polysaccharides esters. Dynamic light scattering, scanning electron microscopy with energy dispersive X-ray spectroscopy, UV-visible absorption spectroscopy, fluorescence spectroscopy, magnetic separation technique and enzyme catalytic assay confirmed the chemical composition of the gold covered superparamagnetic nanoparticles, the binding and activity of the enzyme onto the nanoparticles. Activity data in U/ml confirmed that the immobilized enzyme is approximately 2 times more active than the free enzyme in solution. Such particles can be directed with external magnetic fields for bio-separation and focused towards a medical target for therapeutical as well as bio-sensor applications.
NanoBiotechnology Group, Department of Physics and Nanotechnology, Aalborg University, Skjernvej 4A, Aalborg, Denmark.
Antonietta
A
Parracino
Gnana Prakash
GP
Gajula
Ane Kold
AK
di Gennaro
Maria Teresa
MT
Neves-Petersen
Jens
J
Rafaelsen
Steffen B
SB
Petersen
D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism; D004864:Equipment and Supplies; D006046:Gold / Q000737:chemistry; D008280:Magnetics; D058185:Magnetite Nanoparticles / Q000737:chemistry*; D008958:Models, Molecular; D036103:Nanotechnology / Q000295:instrumentation*; D011487:Protein Conformation; D011549:Pseudomonas / Q000201:enzymology; D013057:Spectrum Analysis; D013499:Surface Properties; D013869:Thiolester Hydrolases / Q000737:chemistry / Q000378:metabolism
D004800:Enzymes, Immobilized; D058185:Magnetite Nanoparticles; D006046:Gold; D013869:Thiolester Hydrolases; C020149:s-formylglutathione hydrolase