Laccase–luminol chemiluminescence system: an investigation of substrate inhibition
Por:
Sánchez-Trasviña C., Galindo-Estrada J.D., Tinoco-Valencia R., Serrano-Carreón L., Rito-Palomares M., Willson R.C., Mayolo-Deloisa K.
Publicada:
1 ene 2023
Ahead of Print:
1 feb 2023
Resumen:
Chemiluminescence (CL) reactions are widely used for the detection and quantification of many types of analytes. Laccase has previously been proposed in CL reactions; however, its light emission behaviour has not been characterized. This study was conducted to characterize the laccase–luminol system, determine its kinetic parameters, and analyze the effects of protein and OH¯ concentration on the CL signal. Laccase from Coriolopsis gallica was combined with different concentrations of luminol (125 nM to 4 mM), and the enzyme kinetics were evaluated using diverse kinetic models. The laccase–luminol system was able to produce CL without an intermediate molecule, but it exhibited substrate-inhibition behaviour. A two-site random model was used and suggested that when the first luminol molecule was bound to the active site, laccase affinity for the second luminol molecule was increased. This inhibition effect could be avoided using a low luminol concentration. At 5 µM luminol concentration, 1 mg/ml (0.13 U) laccase is needed to achieve nearly 90% of the maximum CL signal, suggesting that the available luminol could not bind to all active sites. Furthermore, the concentration of NaOH negatively affected the CL signal. The laccase–luminol system represents an alternative to existing CL systems, with potential uses in molecular detection and quantification. © 2023 John Wiley & Sons Ltd.
Filiaciones:
Sánchez-Trasviña C.:
Tecnologico de Monterrey, The Institute for Obesity Research, Av. Eugenio Garza Sada 2501 Sur, NL, Monterrey, Mexico
Tecnologico de Monterrey, Escuela de Ingeniería y Ciencias, Centro de Biotecnología-FEMSA, Av. Eugenio Garza Sada 2501 Sur, NL, Monterrey, Mexico
Galindo-Estrada J.D.:
Universidad Politécnica del Valle de Toluca, Carretera Toluca-Almoloya de Juárez km. 5.6, Estado de México, Almoloya de Juárez, Mexico
Tinoco-Valencia R.:
Instituto de Biotecnología, Universidad Nacional Autónoma de México, Av. Universidad 2001, Col. Chamilpa, Morelos, Cuernavaca, Mexico
Serrano-Carreón L.:
Instituto de Biotecnología, Universidad Nacional Autónoma de México, Av. Universidad 2001, Col. Chamilpa, Morelos, Cuernavaca, Mexico
Rito-Palomares M.:
Tecnologico de Monterrey, The Institute for Obesity Research, Av. Eugenio Garza Sada 2501 Sur, NL, Monterrey, Mexico
Tecnologico de Monterrey, Escuela de Medicina y Ciencias de la Salud, Av. Morones Prieto 3000 Pte, NL, Monterrey, Mexico
Willson R.C.:
Tecnologico de Monterrey, Escuela de Medicina y Ciencias de la Salud, Av. Morones Prieto 3000 Pte, NL, Monterrey, Mexico
Department of Chemical and Biomolecular Engineering, University of Houston, Houston, TX, United States
Mayolo-Deloisa K.:
Tecnologico de Monterrey, The Institute for Obesity Research, Av. Eugenio Garza Sada 2501 Sur, NL, Monterrey, Mexico
Tecnologico de Monterrey, Escuela de Ingeniería y Ciencias, Centro de Biotecnología-FEMSA, Av. Eugenio Garza Sada 2501 Sur, NL, Monterrey, Mexico
Institute of Materials Science of Barcelona (ICMAB-CSIC), Campus UAB, Bellaterra, Spain
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