Use of Laccase Enzymes As Bio-Receptors for the Organic Dye Methylene Blue in a Surface Plasmon Resonance Biosensor

Overview

Journal Sensors (Basel)

Publisher MDPI

Specialty Biotechnology

Date 2025 Jan 8

PMID 39771744

Authors

Araceli Sanchez-Alvarez

Gabriela Elizabeth Quintanilla-Villanueva

Osvaldo Rodriguez-Quiroz

Melissa Marlene Rodriguez-Delgado

Juan Francisco Villarreal-Chiu

Analia Sicardi-Segade

Donato Luna-Moreno

Affiliations

Soon will be listed here.

Abstract

Methylene blue is a cationic organic dye commonly found in wastewater, groundwater, and surface water due to industrial discharge into the environment. This emerging pollutant is notably persistent and can pose risks to both human health and the environment. In this study, we developed a Surface Plasmon Resonance Biosensor employing a BK7 prism coated with 3 nm chromium and 50 nm of gold in the Kretschmann configuration, specifically for the detection of methylene blue. For the first time, laccases immobilized on a gold surface were utilized as bio-receptors for this organic dye. The enzyme was immobilized using carbodiimide bonds with EDC/NHS crosslinkers, allowing for the analysis of samples with minimal preparation. The method demonstrated validation with a limit of detection (LOD) of 4.61 mg L and a limit of quantification (LOQ) of 15.37 mg L, a working range of 0-100 mg L, and an R value of 0.9614 during real-time analysis. A rainwater sample spiked with methylene blue yielded a recovery rate of 122.46 ± 4.41%. The biosensor maintained a stable signal over 17 cycles and remained effective for 30 days at room temperature.

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