Abstract
The preservation of enzymatic activity is a fundamental requirement for exploiting hybrid nano-bio-conjugates, and the control over protein–nanoparticle interactions, leading to stable and catalytically active hybrids, represents the key for designing new biosensing platforms. In this scenario, surface active maghemite nanoparticles (SAMNs) represent a new class of naked magnetic nanoparticles, displaying peculiar electrocatalytic features and the ability to selectively bind proteins. Recombinant aminoaldehyde dehydrogenase from tomato (SlAMADH1) was used as a model protein, and successfully immobilized by self-assembly on the surface of naked SAMNs, where its enzymatic activity resulted preserved for more than 6 months. The hybrid nanomaterial (SAMN@SlAMADH1) was characterized by UV–Vis spectroscopy, mass spectrometry, and TEM microscopy, and applied for the development of a biosensor for the determination of aminoaldehydes in alcoholic beverages. Measurements were carried out in a low volume electrochemical flow cell comprising a SAMN modified carbon paste electrode for the coulometric determination of the NADH produced during the enzymatic catalysis. The present findings, besides representing the first example of an electrochemical biosensor for aminoaldehydes in an alcoholic matrix, open the door to the use of immobilized enzymes on naked metal oxides nanomaterials for biosensing.
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Abbreviations
- APAL:
-
3-Aminopropionaldehyde diethylacetal
- CP:
-
Carbon paste
- CPE:
-
Carbon paste electrode
- CV:
-
Cyclic voltammetry
- FDR:
-
False discovery rates
- FTIR:
-
Fourier-transform infrared
- His-tag:
-
Histidine-tag
- IDA:
-
Iminodiacetate
- IPTG:
-
Isopropyl β-d-1-thiogalactopyranoside
- LB:
-
Luria–Bertani
- LC:
-
Liquid chromatography
- LOD:
-
Limit of detection
- MGF:
-
Mascot generic format
- MS:
-
Mass spectrometry
- NADH:
-
Nicotinamide adenine dinucleotide
- Q-TOF:
-
Quadrupole time of flight
- RP:
-
Reverse-phase
- RSD:
-
Relative standard deviation
- SAMNs:
-
Surface active maghemite nanoparticles
- SlAMADH1:
-
(Solanum lycopersium) aminoaldehyde dehydrogenase 1
- SCE:
-
Saturated calomel electrode
- SD:
-
Standard deviation
- S/N:
-
Signal/noise
- TEM:
-
Transmission electron microscopy
- TRIS:
-
Tris(hydroxymethyl)aminomethane
- UV–Vis:
-
Ultraviolet–visible
- XRD:
-
X-ray diffraction
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Acknowledgements
The present experimental work was partially funded by Italian Institutional Ministry Grants Cod. 60A06-7411 and 60A06-8055. The authors gratefully acknowledge the University of Padua (Italy), Grant PRAT 2015 (progetti di Ateneo) n. CPDA159850 and the CARIPARO Foundation for the support. The team members from the Czech Republic were supported by Grant No. LO1204 from the Ministry of Education, Youth and Sports. The authors also thank ‘La Sapienza’ University of Rome and Italian MIUR (Ministero dell’Istruzione, dell’Università e della Ricerca). Our gratitude is also due to the “International Polyamine Foundation-ONLUS” for the availability to look up in the Polyamines documentation.
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Magro, M., Baratella, D., Miotto, G. et al. Enzyme self-assembly on naked iron oxide nanoparticles for aminoaldehyde biosensing. Amino Acids 51, 679–690 (2019). https://doi.org/10.1007/s00726-019-02704-7
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DOI: https://doi.org/10.1007/s00726-019-02704-7