Harnessing the Materials Chemistry of Mesoporous Silicon Nanoparticles to Prepare “Armor-Clad” Enzymes

Yi Sheng Lu; Ruhan Fan; Willem Vugs; Lieuwe Biewenga; Ziming Zhou; Sanahan Vijayakumar; Maarten Merkx; Michael J. Sailor

doi:10.1021/acs.chemmater.3c02637

Harnessing the Materials Chemistry of Mesoporous Silicon Nanoparticles to Prepare “Armor-Clad” Enzymes

Yi Sheng Lu, Ruhan Fan, Willem Vugs, Lieuwe Biewenga, Ziming Zhou, Sanahan Vijayakumar, Maarten Merkx, Michael J. Sailor (Corresponding author)

Onderzoeksoutput: Bijdrage aan tijdschrift › Tijdschriftartikel › Academic › peer review

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There is a growing interest in nanomaterials that can encapsulate enzymes while retaining their ability to function within the confines of a nanocage. Here porous silicon nanoparticles (pSiNPs) are evaluated as an enzyme cage, utilizing the aqueous chemistry of silicon to dynamically restructure the mesopore structure, immobilizing, and confining the enzyme. The common bioluminescent reporter enzyme nanoluciferase (Nluc) is used to evaluate two different trapping chemistries, and impacts on the stability and catalytic performance of the enzyme are compared with controls involving free enzyme and enzyme electrostatically adsorbed to a pSiNP host without the use of trapping chemistry. The two chemistries exploited in this study are (1) oxidative trapping, where mild aqueous oxidation of the elemental silicon skeleton in the mesoporous silicon host swells and restructures the pore walls, physically trapping the Nluc payload in a porous SiO₂ matrix, and (2) calcium ion-induced condensation, where localized precipitation of calcium silicate entraps the Nluc protein in a porous silicate matrix. The two trapping chemistries form robust nanoscale cages with substantially smaller pores (9.8 ± 0.4 and 8.8 ± 0.3 nm, respectively) compared to the pSiNP starting material (15.3 ± 1.8 nm), such that the enzyme does not leach from the pSiNPs in aqueous buffer or under assay conditions. Enzyme stability is substantially improved using the two trapping chemistries; the caged materials retain 30-45% activity after heating to 80 °C for 30 min or when exposed to organic solvents; either of these denaturing conditions result in complete or near-complete loss of activity for the free enzyme or for enzyme that is electrostatically adsorbed to pSiNPs. Finally, we explore the potential for the use of the Nluc-encapsulated nanocomposite as a cellular probe by demonstrating the luminescent reporting function of the nanoparticles in HeLa human cell cultures.

Originele taal-2	Engels
Pagina's (van-tot)	10247-10257
Aantal pagina's	11
Tijdschrift	Chemistry of Materials
Volume	35
Nummer van het tijdschrift	23
DOI's	https://doi.org/10.1021/acs.chemmater.3c02637
Status	Gepubliceerd - 12 dec. 2023

Financiering

We thank Dr. Simone Wouters for providing initial support of this project. This research was primarily supported by NSF through the UC San Diego Materials Research Science and Engineering Center (UCSD MRSEC), DMR-2011924. Additional support provided from the NIH through grant R01 AI132413-01 and from the Defense Advanced Research Projects Agency (DARPA) and Naval Information Warfare Center Pacific, (NIWC Pacific) under Contract No. N66001-21-C-4010. Any opinions, findings and conclusions or recommendations expressed in this material are those of the author(s) and do not necessarily reflect the views of the DARPA or NIWC Pacific. The authors acknowledge the use of facilities and instrumentation supported by the National Science Foundation through the UC San Diego Materials Research Science and Engineering Center (UCSD MRSEC) DMR-2011924, and by the San Diego Nanotechnology Infrastructure (SDNI) of UCSD, a member of the National Nanotechnology Coordinated Infrastructure, which is supported by the National Science Foundation (Grant ECCS-2025752).

Financiers	Financiernummer
National Science Foundation(NSF)	ECCS-2025752
National Institutes of Health, NIH	R01 AI132413-01
Defense Advanced Research Projects Agency
Naval Information Warfare Center Pacific	N66001-21-C-4010
University of California at San Diego	DMR-2011924

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abstract = "There is a growing interest in nanomaterials that can encapsulate enzymes while retaining their ability to function within the confines of a nanocage. Here porous silicon nanoparticles (pSiNPs) are evaluated as an enzyme cage, utilizing the aqueous chemistry of silicon to dynamically restructure the mesopore structure, immobilizing, and confining the enzyme. The common bioluminescent reporter enzyme nanoluciferase (Nluc) is used to evaluate two different trapping chemistries, and impacts on the stability and catalytic performance of the enzyme are compared with controls involving free enzyme and enzyme electrostatically adsorbed to a pSiNP host without the use of trapping chemistry. The two chemistries exploited in this study are (1) oxidative trapping, where mild aqueous oxidation of the elemental silicon skeleton in the mesoporous silicon host swells and restructures the pore walls, physically trapping the Nluc payload in a porous SiO2 matrix, and (2) calcium ion-induced condensation, where localized precipitation of calcium silicate entraps the Nluc protein in a porous silicate matrix. The two trapping chemistries form robust nanoscale cages with substantially smaller pores (9.8 ± 0.4 and 8.8 ± 0.3 nm, respectively) compared to the pSiNP starting material (15.3 ± 1.8 nm), such that the enzyme does not leach from the pSiNPs in aqueous buffer or under assay conditions. Enzyme stability is substantially improved using the two trapping chemistries; the caged materials retain 30-45% activity after heating to 80 °C for 30 min or when exposed to organic solvents; either of these denaturing conditions result in complete or near-complete loss of activity for the free enzyme or for enzyme that is electrostatically adsorbed to pSiNPs. Finally, we explore the potential for the use of the Nluc-encapsulated nanocomposite as a cellular probe by demonstrating the luminescent reporting function of the nanoparticles in HeLa human cell cultures.",

author = "Lu, {Yi Sheng} and Ruhan Fan and Willem Vugs and Lieuwe Biewenga and Ziming Zhou and Sanahan Vijayakumar and Maarten Merkx and Sailor, {Michael J.}",

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Harnessing the Materials Chemistry of Mesoporous Silicon Nanoparticles to Prepare “Armor-Clad” Enzymes

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