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

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)₂AuNPs, PEG(SH)₂Au/F₃O₄NPs, CysAuNPs, and CysAu/F₃O₄NPs 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.

SUBMITTER: Belleti E

PROVIDER: S-EPMC8493054 | biostudies-literature | 2021 Nov

REPOSITORIES: biostudies-literature

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

Belleti Elisângela E Bevilaqua Vanessa R VR Brito Adrianne M M AMM Modesto Diego A DA Lanfredi Alexandre J C AJC Viviani Vadim R VR Nantes-Cardoso Iseli L IL

Photochemical & photobiological sciences : Official journal of the European Photochemistry Association and the European Society for Photobiology 20211006 11

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)2AuNPs, PEG(SH)2Au/F3O4NPs, CysAuNPs, and CysAu/F3 ...[more]

PMID: 34613602

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Project description:The unique features of Metal-Organic Frameworks (MOFs), including structural flexibility, high surface area, and variable pore size, have drawn attention in cancer therapy. However, despite advances in surface functionalization, engineering structural features, and porosity, achieving controlled release, stability, scalability, and toxicity remains a challenge. The current study reports gold nanoparticle (AuNP) encapsulated dual metal-organic frameworks (MOFs) comprising zeolitic imidazolate (ZIF8) and cobalt-imidazole (ZIF67) by a simple precipitation method for dual drug delivery applications. This combination associates the advantages of AuNPs and MOFs, creating a potent platform for cancer theranostics that combines diagnosis and treatment into one unit. The synthesized composite (AuNPs@ZIF-8/ZIF-67) is functionalized with Folic acid (FA) and loaded with the anticancer agents Curcumin (C) and 5-fluorouracil (5-FU) for co-drug delivery The synthesized composites, namely Au/ZIF8, Au/ZIF8/ZIF67/FA, Au/ZIF8/ZIF67/FA/5-FU, and Au/ZIF8/ZIF67/FA/5-FU/C were characterized using diverse analytical techniques such as FESEM, XRD, FTIR, TEM, and BET. The characterization methods showed that the hybrid MOF structure was stable and intact after AuNP encapsulation and drug loading. The dual MOF composite exhibits a better affinity for loading C and 5-FU with 60% and 40% drug loading capacity, respectively. The simultaneous drug release studies suggest that AuNPs@ZIF-8/ZIF-67 are more responsive to the acidic pH and show a higher cumulative drug release of 5FU and C at the lower value of pH 5. For further validation, the release kinetics data were fitted into the Korsmeyer-Peppas model in the current study. The observed value of n which is less than 0.5 suggests the pseudo-Fickian diffusion mechanism for drug release, demonstrating long-term release of 5FU and C from Au/ZIF8/ZIF67/FA/5-FU/C. The targeted drug delivery system is anticipated to display synergistic therapeutic efficacy from the combined effect of the two anticancer agents and the pH-responsive nature of ZIF systems.

Dataset Information

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

Publications

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

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