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

Abstract

Graphic abstract Bioluminescent gold nanoparticles (AuNPs) were synthesized in situ using dithiol-terminated polyethylene glycol (PEG(SH) 2 ) as reducer and stabilizing agents. Hybrid Au/F 3 O 4 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. Supplementary Information The online version contains supplementary material available at 10.1007/s43630-021-00111-0.

Calculation of the Localized Surface Plasmon Resonances of Au Nanoparticles Embedded in NiO

“…The scope of this work is to controversially show the appearance of LSPRs generated with the aid of software analysis. This study focuses on the noble metal Au, a promising material according to recent studies, due to the numerous technological applications; it participates, characteristically mentioning the diode and biochemical devices, the bioluminescence and biosensing applications, the electromechanical sensors, the photocatalytic activity, the Metal-Organic Framework, the photocatalytic mechanism for the solar, and photovoltaic production [11][12][13][14][15][16][17][18].…”

Label-Free and Bioluminescence-Based Nano-Biosensor for ATP Detection

“…In another study, Ebrahimi et al (2015) showed that P. pyralis luciferase could be electrostatically immobilized on functionalized magnetic nanoparticles (MNPs) of His-tag with Ni 2+ /Cu 2+ ion immobilization, which decreased the affinity for ATP while increasing the affinity for luciferin without affecting thermal stability or longevity [ 22 ]. The effect of gold nanoparticles on the purified recombinant green-emitting click beetle luciferase and red-emitting railroad worm luciferase has also been reported [ 23 ]. The fluorescent metal nanoclusters have tens of atoms and behave like molecules without a featured surface plasmon resonance peak in the visible region.…”