The identification of new strategies to improve the stability of proteins is of utmost importance for a number of applications, from biosensing to biocatalysis. Metal-organic frameworks (MOFs) have been shown as a versatile host platform for the immobilization of proteins, with the potential to protect proteins in harsh conditions. In this work, a new thermostable luciferase mutant has been selected as a bioluminescent protein model to investigate the suitability of MOFs to improve its stability and prompt its applications in real-world applications, for example, ATP detection in portable systems. The luciferase has been immobilized onto zeolitic imidazolate framework-8 (ZIF-8) to obtain a bioluminescent biocomposite with enhanced performance. The biocomposite ZIF-8@luc has been characterized in harsh conditions (e.g., high temperature, non-native pH, etc.). Bioluminescence properties confirmed that MOF enhanced the luciferase stability at acidic pH, in the presence of organic solvents, and at -20 degrees C. To assess the feasibility of this approach, the recyclability, storage stability, precision, and Michaelis-Menten constants (Km) for ATP and D-luciferin have been also evaluated. As a proof of principle, the suitability for ATP detection was investigated and the biocomposite outperformed the free enzyme in the same experimental conditions, achieving a limit of detection for ATP down to 0.2 fmol.

Martínez-Pérez-Cejuela, H., Gregucci, D., Calabretta, M.M., Simó-Alfonso, E.F., Herrero-Martínez, J.M., Michelini, E. (2023). Novel Nanozeolitic Imidazolate Framework (ZIF-8)–Luciferase Biocomposite for Nanosensing Applications. ANALYTICAL CHEMISTRY, 95(4), 2540-2547 [10.1021/acs.analchem.2c05001].

Novel Nanozeolitic Imidazolate Framework (ZIF-8)–Luciferase Biocomposite for Nanosensing Applications

Gregucci, Denise;Calabretta, Maria Maddalena;Michelini, Elisa
Ultimo
2023

Abstract

The identification of new strategies to improve the stability of proteins is of utmost importance for a number of applications, from biosensing to biocatalysis. Metal-organic frameworks (MOFs) have been shown as a versatile host platform for the immobilization of proteins, with the potential to protect proteins in harsh conditions. In this work, a new thermostable luciferase mutant has been selected as a bioluminescent protein model to investigate the suitability of MOFs to improve its stability and prompt its applications in real-world applications, for example, ATP detection in portable systems. The luciferase has been immobilized onto zeolitic imidazolate framework-8 (ZIF-8) to obtain a bioluminescent biocomposite with enhanced performance. The biocomposite ZIF-8@luc has been characterized in harsh conditions (e.g., high temperature, non-native pH, etc.). Bioluminescence properties confirmed that MOF enhanced the luciferase stability at acidic pH, in the presence of organic solvents, and at -20 degrees C. To assess the feasibility of this approach, the recyclability, storage stability, precision, and Michaelis-Menten constants (Km) for ATP and D-luciferin have been also evaluated. As a proof of principle, the suitability for ATP detection was investigated and the biocomposite outperformed the free enzyme in the same experimental conditions, achieving a limit of detection for ATP down to 0.2 fmol.
2023
Martínez-Pérez-Cejuela, H., Gregucci, D., Calabretta, M.M., Simó-Alfonso, E.F., Herrero-Martínez, J.M., Michelini, E. (2023). Novel Nanozeolitic Imidazolate Framework (ZIF-8)–Luciferase Biocomposite for Nanosensing Applications. ANALYTICAL CHEMISTRY, 95(4), 2540-2547 [10.1021/acs.analchem.2c05001].
Martínez-Pérez-Cejuela, Héctor; Gregucci, Denise; Calabretta, Maria Maddalena; Simó-Alfonso, Ernesto Francisco; Herrero-Martínez, José Manuel; Micheli...espandi
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11585/959207
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