Generally, enzyme immobilization on nanoparticles leads to nano-conjugates presenting partially preserved, or even absent, biological properties. Notwithstanding, recent research demonstrated that the coupling to nanomaterials can improve the activity of immobilized enzymes. Herein, xanthine oxidase (XO) was immobilized by self-assembly on peculiar naked iron oxide nanoparticles (surface active maghemite nanoparticles, SAMNs). The catalytic activity of the nanostructured conjugate (SAMN@XO) was assessed by optical spectroscopy and compared to the parent enzyme. SAMN@XO revealed improved catalytic features with respect to the parent enzyme and was applied for the electrochemical studies of xanthine. The present example supports the nascent knowledge concerning protein conjugation to nanoparticle as a means for the modulation of biological activity.

Department of Chemistry and Biochemistry Institute of Bioscience Universidade Estadual Paulista Botucatu SP 18618 000 Brazil

Department of Comparative Biomedicine and Food Science University of Padua Agripolis Viale dell'Università 16 35020 Legnaro Italy

Department of Geoscience University of Padua via G Gradenigo 6 35131 Padua Italy

Regional Centre of Advanced Technologies and Materials Palacky University in Olomouc Slechtitelu 27 783 71 Olomouc Czech Republic

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Rana S., Yeh Y.C., Rotello V.M. Engineering the nanoparticle-protein interface: Applications and possibilities. Curr. Opin. Chem. Biol. 2010;14:828–834. doi: 10.1016/j.cbpa.2010.10.001. PubMed DOI PMC

Misson M., Zhang H., Jin B. Nanobiocatalyst advancements and bioprocessing applications. J. R. Soc. Interface. 2015;12:20140891. doi: 10.1098/rsif.2014.0891. PubMed DOI PMC

Mahmoudi M., Bertrand N., Zope H., Farokzad O.C. Emerging understanding of the protein corona at the nano-bio interfaces. Nano Today. 2016;11:817–832. doi: 10.1016/j.nantod.2016.10.005. DOI

Mahmoudi M., Lynch I., Ejtehadi M.R., Monopoli M.P., BaldelliBombelli F., Laurent S. Protein−nanoparticle interactions: Opportunities and challenges. Chem. Rev. 2011;111:5610–5637. doi: 10.1021/cr100440g. PubMed DOI

Burda C., Chen X., Narayanan R., El-Sayed M.A. Chemistry and properties of nanocrystals of different shapes. Chem. Rev. 2005;105:1025–1102. doi: 10.1021/cr030063a. PubMed DOI

Garcia J., Zhang Y., Taylor H., Cespedes O., Webb M.E., Zhou D. Multilayer enzyme-coupled magnetic nanoparticles as efficient, reusable biocatalysts and biosensors. Nanoscale. 2011;3:3721–3730. doi: 10.1039/c1nr10411j. PubMed DOI

Niemirowicz K., Markiewicz K.H., Wilczewska A.Z., Car H. Magnetic nanoparticles as new diagnostic tools in medicine. Adv. Med. Sci. 2012;57:196–207. doi: 10.2478/v10039-012-0031-9. PubMed DOI

Johnson B.J., Algar W.R., Malanoski A.P., Ancona M.G., Medintz I.L. Understanding enzymatic acceleration at nanoparticle interfaces: Approaches and challenges. Nano Today. 2014;9:102–131. doi: 10.1016/j.nantod.2014.02.005. DOI

Ding S., Cargill A.A., Medintz I.L., Claussen J.C. Increasing the activity of immobilized enzymes with nanoparticle conjugation. Curr. Opin. Biotechnol. 2015;34:242–250. doi: 10.1016/j.copbio.2015.04.005. PubMed DOI

Netto C.G.C.M., Toma H.E., Andrade L.H. Superparamagnetic nanoparticles as versatile carriers and supporting materials for enzymes. J. Mol. Catal. B Enzym. 2013;85–86:71–92. doi: 10.1016/j.molcatb.2012.08.010. DOI

Zhang H.L., Lai G.S., Han D.Y., Yu A.M. An amperometric hydrogen peroxide biosensor based on immobilization of horseradish peroxidase on an electrode modified with magnetic dextran microspheres. Anal. Bioanal. Chem. 2008;390:971–977. doi: 10.1007/s00216-007-1748-3. PubMed DOI

Magro M., Baratella D., Miotto G., Frömmel J., Šebela M., Kopečná M., Agostinelli E., Vianello F. Enzyme self-assembly on naked iron oxide nanoparticles for aminoaldehyde biosensing. Amino Acids. 2019;51:679–690. doi: 10.1007/s00726-019-02704-7. PubMed DOI

Magro M., Baratella D., Bonaiuto E., de Almeida Roger J., Vianello F. New Perspectives on Biomedical Applications of Iron Oxide Nanoparticles. Curr. Med. Chem. 2018;25:540–555. doi: 10.2174/0929867324666170616102922. PubMed DOI

Magro M., Sinigaglia G., Nodari L., Tucek J., Polakova K., Zdenĕk M., Cardillo S., Salviulo G., Russo U., Zboril R., et al. Charge binding of rhodamine derivative to OH- stabilized nanomaghemite: Universal nanocarrier for construction of magneto fluorescent biosensors. Acta Biomater. 2012;8:2068–2076. doi: 10.1016/j.actbio.2012.02.005. PubMed DOI

Pundir C.S., Devi R. Biosensing methods for xanthine determination: A review. Enzym. Microb. Technol. 2014;57:55–62. doi: 10.1016/j.enzmictec.2013.12.006. PubMed DOI

Shintani H. Determination of xanthine oxidase. Pharm. Anal. Acta S. 2013;7:004. doi: 10.4172/2153-2435.S7-004. DOI

Fersht A. Enzyme Structure and Mechanism. 1st ed. W.H. Freeman & Co.; New York, NY, USA: 1984.

Bradford M.M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 1976;72:248–252. doi: 10.1016/0003-2697(76)90527-3. PubMed DOI

Venerando R., Miotto G., Magro M., Dallan M., Baratella D., Bonaiuto E., Zboril R., Vianello F. Magnetic nanoparticles with covalently bound self-assembled protein corona for advanced biomedical applications. J. Phys. Chem. C. 2013;117:20320–20331. doi: 10.1021/jp4068137. DOI

Magro M., Faralli A., Baratella D., Bertipaglia I., Giannetti S., Salviulo G., Zboril R., Vianello F. Avidin functionalized maghemite nanoparticles and their application for recombinant human biotinyl-SERCA purification. Langmuir. 2012;28:15392–15401. doi: 10.1021/la303148u. PubMed DOI

Enroth C., Eger B.T., Okamoto K., Nishino T., Nishino T., Pai E.F. Crystal structures of bovine milk xanthine dehydrogenase and xanthine oxidase: Structure-based mechanism of conversion. Proc. Natl. Acad. Sci. USA. 2000;97:10723–10728. doi: 10.1073/pnas.97.20.10723. PubMed DOI PMC

Battelli M.G., Lorenzoni E., Stripe F. Milk xanthine oxidase type D (dehydrogenase) and type O (oxidase). Purification, interconversion and some properties. Biochem. J. 1973;131:191–198. doi: 10.1042/bj1310191. PubMed DOI PMC

Stein B.W., Kirk M.L. Electronic structure contributions to reactivity in xanthine oxidase family enzymes. J. Biol. Inorg. Chem. 2015;20:183–194. doi: 10.1007/s00775-014-1212-8. PubMed DOI PMC

Cerqueira N.M., Pakhira B., Sarkar S. Theoretical studies on mechanisms of some Mo enzymes. J. Biol. Inorg. Chem. 2015;20:323–335. doi: 10.1007/s00775-015-1237-7. PubMed DOI

Bielski B.H.J., Allen A.O. Mechanism of the disproportionation of superoxide radicals. J. Phys. Chem. 1977;81:1048–1050. doi: 10.1021/j100526a005. DOI

Johnson K.A. Role of induced fit in enzyme specificity: A molecular forward/reverse switch. J. Biol. Chem. 2008;283:26297–26301. doi: 10.1074/jbc.R800034200. PubMed DOI PMC

Chong A.S.M., Zhao X.S. Design of large-pore mesoporous materials for immobilization of penicillin G acylase biocatalyst. Catal. Today. 2004;93-95:293–299. doi: 10.1016/j.cattod.2004.06.064. DOI

Pan C., Hu B., Li W., Sun Y., Ye H., Zeng X. Novel and efficient method for immobilization and stabilization of β-D-galactosidase by covalent attachment onto magnetic Fe3O4–chitosan nanoparticles. J. Mol. Catal. B Enzym. 2009;61:208–215. doi: 10.1016/j.molcatb.2009.07.003. DOI

Lynch I., Dawson K.A. Protein-nanoparticle interactions. Nano Today. 2008;3:40–47. doi: 10.1016/S1748-0132(08)70014-8. DOI

Magro M., Zaccarin M., Miotto G., Da Dalt L., Baratella D., Fariselli P., Gabai G., Vianello F. Analysis of hard protein corona composition on selective iron oxide nanoparticles by MALDI-TOF mass spectrometry: Identification and amplification of a hidden mastitis biomarker in milk proteome. Anal. Bioanal. Chem. 2018;410:2949–2959. doi: 10.1007/s00216-018-0976-z. PubMed DOI

Miller J.N., Miller J.C. Statistics and Chemometrics for Analytical Chemistry. 6th ed. Pearson/Prentice Hall; Harlow, UK: 2010.

Nakatani H.S., Santos L.V.D., Pelegrine C.P., Gomes M., Matsushita M., Souza N.E.D., Visentainer J.V. Biosensor based on xanthine oxidase for monitoring hypoxanthine in fish meat. Am. J. Biochem. Biotechnol. 2005;1:85–89. doi: 10.3844/ajbbsp.2005.85.89. DOI

Cubukcu M., Timur S., Anik U. Examination of performance of glassy carbon paste electrode modified with gold nanoparticle and xanthine oxidase for xanthine and hypoxanthine detection. Talanta. 2007;74:434–439. doi: 10.1016/j.talanta.2007.07.039. PubMed DOI

Devi R., Thakur M., Pundir C.S. Construction and application of an amperometric xanthine biosensor based on zinc oxide nanoparticles–polypyrrole composite film. Biosens. Bioelectron. 2011;26:3420–3423. doi: 10.1016/j.bios.2011.01.014. PubMed DOI

Anik U., Çevik S. Double-walled carbon nanotube based carbon pastel electrode as xanthine biosensor. Microchim. Acta. 2009;166:209–221. doi: 10.1007/s00604-009-0190-y. DOI

Dervisevic M., Dervisevic E., Azak H., Çevik E., Şenel M., Yildiz H.B. Novel amperometric xanthine biosensor based on xanthine oxidase immobilized on electrochemically polymerized 10-[4H-dithieno(3,2-b:2′,3′-d) pyrrole-4-yl]decane-1-amine film. Sens. Actuators B Chem. 2016;225:181–187. doi: 10.1016/j.snb.2015.11.043. DOI

Dalkiran B., Erden P.E., Kiliç E. Amperometric biosensors based on carboxylated multi-walled carbon nanotubes-metal oxide nanoparticles-7,7,8,8-tetracyanoquinodimethane composite for the determination of xanthine. Talanta. 2017;167:286–295. doi: 10.1016/j.talanta.2017.02.021. PubMed DOI

Pei J., Li X.Y. Xanthine and hypoxanthine sensors based on xanthine oxidase immobilized on a CuPtCl6 chemically modified electrode and liquid chromatography electrochemical detection. Anal. Chem. Acta. 2000;414:205–213. doi: 10.1016/S0003-2670(00)00775-3. DOI