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

Autor(es)

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]

Afiliação do(s) autor(es)

^[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

• Tipo de documento: Artigo Científico
• Fonte: PHOTOCHEMICAL & PHOTOBIOLOGICAL SCIENCES; v. 20, n. 11 OCT 2021.
• Citações Web of Science: 0

Resumo

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)

• Processo FAPESP: 10/05426-8 - Bioluminescência de artrópodes: diversidade biológica em biomas brasileiros; origem bioquímica; evolução estrutural/funcional de luciferases; diferenciação molecular das lanternas; aplicações biotecnológicas, ambientais e educacionais
• Beneficiário: Vadim Viviani
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 17/02317-2 - Interfaces em materiais: propriedades eletrônicas, magnéticas, estruturais e de transporte
• Beneficiário: Adalberto Fazzio
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 15/17688-0 - Nanoestruturas Metálicas Associadas a Porfirinas, Citocromo c e Azul de Metileno: Estudos Fundamentais para Aplicações em Sistema Biológicos e Energia
• Beneficiário: Iseli Lourenço Nantes Cardoso
• Modalidade de apoio: Auxílio à Pesquisa - Regular