Fluorescence detection of laccases activity by the photoinduced electron transfer (PET) process

J Biol Inorg Chem. 2020 Feb;25(1):151-159. doi: 10.1007/s00775-019-01748-0. Epub 2019 Dec 12.

Abstract

Laccases play a vital role in some physiological processes, for example in morphogenesis, carbon cycle, and defense against parasitism. So, designing a high-sensitivity accurate method is essential for researchers. In this study, a simple fluorescence method based on the function of carbon nitride (g-C3N4) by dopamine is synthesized. For the design of this sensor, carbon nitride (g-C3N4) is initially synthesis by using a simple method, which is carried out by heating melamine at 550 °C for 3 h and modifying it with dopamine by a linker such as glutaraldehyde. However, the g-C3N4-Dopa produced by this method, with an excitation wavelength of 330 nm, has a fluorescence emission at 466 nm. When laccase and g-C3N4-Dopa were mixed, dopamine with redox property was oxidized to dopaquinone; this causes the phenomenon of photoinduced electron transfer (PET) process between g-C3N4 and the dopaquinone. Hence, fluorescence quenching occurs due to this phenomenon. As a result of these discussions, a sensor for the laccase activity was designed based on the fluorescence quenching degree, supporting a linear range of 0.0-400.0 U L-1 with the detection limit of 2.0 U L-1. Using this sensor, the activity of the laccase enzyme in the human serum samples is measured. Dopamine-functionalized carbon nitride was prepared and utilized for the highly sensitive detection of laccases activity.

Keywords: Dopamine; Fluorescence; Graphitic carbon nitride; Laccases activity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Dopamine / chemistry
  • Electron Transport
  • Laccase / metabolism*
  • Light
  • Limit of Detection
  • Polyporaceae / enzymology
  • Spectrum Analysis / methods

Substances

  • Laccase
  • Dopamine

Supplementary concepts

  • Trametes versicolor