Synthesis of bioluminescent gold nanoparticle-luciferase hybrid systems for technological applications

Author(s)

Belleti, Elisangela ^[1] ; Bevilaqua, Vanessa R. ^[2] ; Brito, Adrianne M. M. ^[1] ; Modesto, Diego A. ^[1] ; Lanfredi, Alexandre J. C. ^[1] ; Viviani, Vadim R. ^{[3, 1]} ; Nantes-Cardoso, Iseli L. ^[1]

Affiliation

^[1] Univ Fed ABC UFABC, Av Estados 501, BR-09210580 Santo Andre, SP - Brazil

^[2] Grad Sch Evolut Genet & Mol Biol UFSCar, Sao Carlos, SP - Brazil

^[3] Grad Sch Biotechnol & Environm Monitoring UFSCar, Sorocaba, SP - Brazil

• Document type: Journal article
• Source: PHOTOCHEMICAL & PHOTOBIOLOGICAL SCIENCES; v. 20, n. 11 OCT 2021.
• Web of Science Citations: 0

Abstract

Bioluminescent gold nanoparticles (AuNPs) were synthesized in situ using dithiol-terminated polyethylene glycol (PEG(SH)(2)) as reducer and stabilizing agents. Hybrid Au/F3O4 nanoparticles were also produced in a variation of synthesis, and both types of nanostructures had the polymer capping replaced by l-cysteine (Cys). The four types of nanoparticles, PEG(SH)(2)AuNPs, PEG(SH)(2)Au/F(3)O(4)NPs, CysAuNPs, and CysAu/F(3)O(4)NPs were associated with purified recombinant Pyrearinus termitilluminans green emitting click beetle luciferase (PyLuc) and Phrixotrix hirtus (RELuc) red-emitting railroad worm luciferase. Enzyme association with PEG(SH)(2) was also investigated as a control. Luciferases were chosen because they catalyze bioluminescent reactions used in a wide range of bioanalytical applications, including ATP assays, gene reporting, high-throughput screening, bioluminescence imaging, biosensors and other bioluminescence-based assays. The immobilization of PyLuc and RELuc promoted partial suppression of the enzyme luminescence activity in a functionalization-dependent way. Association of PyLuc and RELuc with AuNPs increased the enzyme operational stability in relation to the free enzyme, as evidenced by the luminescence intensity from 0 to 7 h after substrate addition. The stability of the immobilized enzymes was also functionalization-dependent and the association with CysAuNPs was the condition that combined more sustained luminescent activity with a low degree of luminescence quenching. The higher enzymatic stability and sustained luminescence of luciferases associated with nanoparticles may improve the applicability of bioluminescence for bioimaging and biosensing purposes. Graphic abstract (AU)

• FAPESP's process: 10/05426-8 - Arthropod bioluminescence: biological diversity in Brazilian biomes, biochemical origin, structural/functional evolution of luciferases, molecular differentiation of lanterns, biotechnological, environmental and educational applications
• Grantee: Vadim Viviani
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 17/02317-2 - Interfaces in materials: electronic, magnetic, structural and transport properties
• Grantee: Adalberto Fazzio
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 15/17688-0 - Metallic Nanostructures Associated to Porphyrins, Cytochrome c and Methylene Blue: Fundamental Studies for Applications for Biological Systems and Energy
• Grantee: Iseli Lourenço Nantes Cardoso
• Support Opportunities: Regular Research Grants