Colorimetric assays for biodegradation of polycyclic aromatic hydrocarbons by fungal laccases

J Biomol Screen. 2002 Dec;7(6):547-53. doi: 10.1177/1087057102238629.

Abstract

Polycyclic aromatic hydrocarbons (PAHs) are highly toxic organic pollutants widely distributed in terrestrial and aquatic environments. In the present work, 2 colorimetric assays for laccase-catalyzed degradation of PAHs were developed based on studies of the oxidation of 12 aromatic hydrocarbons by fungal laccases from Trametes versicolor and Myceliophthora thermophila. Using a sodium borohydride water-soluble solution, the authors could reduce the single product of laccase-catalyzed anthracene biooxidation into the orange-colored 9,10-anthrahydroquinone, which is quantifiable spectrophotometrically. An assay using polymeric dye (Poly R-478) as a surrogate substrate for lignin degradation by laccase in the presence of mediator is also presented. The decolorization of Poly R-478 was correlated to the oxidation of PAHs mediated by laccases. This demonstrates that a ligninolytic indicator such as Poly R-478 can be used to screen for PAH-degrading laccases; it will also be useful in screening mutant libraries in directed evolution experiments. Poly R-478 is stable and readily soluble. It has a high extinction coefficient and low toxicity toward white rot fungi, yeast, and bacteria, which allow its application in a solid-phase assay format.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Anthracenes / chemistry
  • Anthracenes / metabolism
  • Anthraquinones / chemistry
  • Anthraquinones / metabolism
  • Biodegradation, Environmental
  • Colorimetry / methods*
  • Coloring Agents / chemistry
  • Fungal Proteins / metabolism*
  • Laccase / metabolism*
  • Oxidation-Reduction
  • Polycyclic Aromatic Hydrocarbons / metabolism*
  • Polymers / chemistry

Substances

  • 9,10-anthrahydroquinone
  • Anthracenes
  • Anthraquinones
  • Coloring Agents
  • Fungal Proteins
  • Polycyclic Aromatic Hydrocarbons
  • Polymers
  • 9,10-anthraquinone
  • poly R-478
  • Laccase
  • anthracene