Skip to main content
PMC home page
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1992 Sep;58(9):3110–3116. doi: 10.1128/aem.58.9.3110-3116.1992

A new assay for lignin-type peroxidases employing the dye azure B.

F S Archibald 1
PMCID: PMC183056  PMID: 1444425

Abstract

The discovery in 1983 of fungal "ligninases" capable of catalyzing the peroxidation of nonphenolic aromatic lignin components has been seen as a major advance in understanding how certain basidiomycete fungi can completely degrade lignin. The ability of these lignin-type peroxidases to covert millimolar concentrations of veratryl alcohol to veratraldehyde, indicated by a change in the A310 of veratraldehyde, has become the standard assay for routine quantitation of LP activity. A new assay based on the oxidation of micromolar concentrations of the dye Azure B is presented. Although it is as simple and rapid as the veratryl alcohol assay, it appears to overcome some of the shortcomings of that assay. In particular, interference from UV- and short-wavelength visible-light-absorbing materials is greatly reduced and assay specificity is improved.

Full text

PDF
3114

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Anliker R. Ecotoxicology of dyestuffs-a joint effort by industry. Ecotoxicol Environ Saf. 1979 Mar;3(1):59–74. doi: 10.1016/0147-6513(79)90060-5. [DOI] [PubMed] [Google Scholar]
  2. Archibald F. S. Lignin Peroxidase Activity Is Not Important in Biological Bleaching and Delignification of Unbleached Kraft Pulp by Trametes versicolor. Appl Environ Microbiol. 1992 Sep;58(9):3101–3109. doi: 10.1128/aem.58.9.3101-3109.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Beauchamp C., Fridovich I. A mechanism for the production of ethylene from methional. The generation of the hydroxyl radical by xanthine oxidase. J Biol Chem. 1970 Sep 25;245(18):4641–4646. [PubMed] [Google Scholar]
  4. Bourbonnais R., Paice M. G. Veratryl alcohol oxidases from the lignin-degrading basidiomycete Pleurotus sajor-caju. Biochem J. 1988 Oct 15;255(2):445–450. doi: 10.1042/bj2550445. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cheton P. L., Archibald F. S. Manganese complexes and the generation and scavenging of hydroxyl free radicals. Free Radic Biol Med. 1988;5(5-6):325–333. doi: 10.1016/0891-5849(88)90104-9. [DOI] [PubMed] [Google Scholar]
  6. Cohen G., Cederbaum A. I. Microsomal metabolism of hydroxyl radical scavenging agents: relationship to the microsomal oxidation of alcohols. Arch Biochem Biophys. 1980 Feb;199(2):438–447. doi: 10.1016/0003-9861(80)90300-8. [DOI] [PubMed] [Google Scholar]
  7. Cripps C., Bumpus J. A., Aust S. D. Biodegradation of azo and heterocyclic dyes by Phanerochaete chrysosporium. Appl Environ Microbiol. 1990 Apr;56(4):1114–1118. doi: 10.1128/aem.56.4.1114-1118.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Glenn J. K., Gold M. H. Decolorization of Several Polymeric Dyes by the Lignin-Degrading Basidiomycete Phanerochaete chrysosporium. Appl Environ Microbiol. 1983 Jun;45(6):1741–1747. doi: 10.1128/aem.45.6.1741-1747.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Glenn J. K., Morgan M. A., Mayfield M. B., Kuwahara M., Gold M. H. An extracellular H2O2-requiring enzyme preparation involved in lignin biodegradation by the white rot basidiomycete Phanerochaete chrysosporium. Biochem Biophys Res Commun. 1983 Aug 12;114(3):1077–1083. doi: 10.1016/0006-291x(83)90672-1. [DOI] [PubMed] [Google Scholar]
  10. Gold M. H., Kuwahara M., Chiu A. A., Glenn J. K. Purification and characterization of an extracellular H2O2-requiring diarylpropane oxygenase from the white rot basidiomycete, Phanerochaete chrysosporium. Arch Biochem Biophys. 1984 Nov 1;234(2):353–362. doi: 10.1016/0003-9861(84)90280-7. [DOI] [PubMed] [Google Scholar]
  11. Kenten R. H., Mann P. J. The oxidation of manganese by peroxidase systems. Biochem J. 1950 Jan;46(1):67–73. doi: 10.1042/bj0460067. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kersten P. J., Kalyanaraman B., Hammel K. E., Reinhammar B., Kirk T. K. Comparison of lignin peroxidase, horseradish peroxidase and laccase in the oxidation of methoxybenzenes. Biochem J. 1990 Jun 1;268(2):475–480. doi: 10.1042/bj2680475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kersten P. J., Tien M., Kalyanaraman B., Kirk T. K. The ligninase of Phanerochaete chrysosporium generates cation radicals from methoxybenzenes. J Biol Chem. 1985 Mar 10;260(5):2609–2612. [PubMed] [Google Scholar]
  14. Lawrence G. D., Cohen G. In vivo production of ethylene from 2-keto-4-methylthiobutyrate in mice. Biochem Pharmacol. 1985 Sep 15;34(18):3231–3236. doi: 10.1016/0006-2952(85)90339-9. [DOI] [PubMed] [Google Scholar]
  15. Perez J., Jeffries T. W. Mineralization of C-Ring-Labeled Synthetic Lignin Correlates with the Production of Lignin Peroxidase, not of Manganese Peroxidase or Laccase. Appl Environ Microbiol. 1990 Jun;56(6):1806–1812. doi: 10.1128/aem.56.6.1806-1812.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Ramachandra M., Crawford D. L., Hertel G. Characterization of an extracellular lignin peroxidase of the lignocellulolytic actinomycete Streptomyces viridosporus. Appl Environ Microbiol. 1988 Dec;54(12):3057–3063. doi: 10.1128/aem.54.12.3057-3063.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Sariaslani F. S., Beale J. M., Jr, Rosazza J. P. Oxidation of rotenone by Polyporus anceps laccase. J Nat Prod. 1984 Jul-Aug;47(4):692–697. doi: 10.1021/np50034a021. [DOI] [PubMed] [Google Scholar]
  18. Tien M., Kirk T. K., Bull C., Fee J. A. Steady-state and transient-state kinetic studies on the oxidation of 3,4-dimethoxybenzyl alcohol catalyzed by the ligninase of Phanerocheate chrysosporium Burds. J Biol Chem. 1986 Feb 5;261(4):1687–1693. [PubMed] [Google Scholar]
  19. Tien M., Kirk T. K. Lignin-Degrading Enzyme from the Hymenomycete Phanerochaete chrysosporium Burds. Science. 1983 Aug 12;221(4611):661–663. doi: 10.1126/science.221.4611.661. [DOI] [PubMed] [Google Scholar]
  20. Tien M., Kirk T. K. Lignin-degrading enzyme from Phanerochaete chrysosporium: Purification, characterization, and catalytic properties of a unique H(2)O(2)-requiring oxygenase. Proc Natl Acad Sci U S A. 1984 Apr;81(8):2280–2284. doi: 10.1073/pnas.81.8.2280. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Yang S. F. Further studies on ethylene formation from alpha-keto-gamma-methylthiobutyric acid or beta-methylthiopropionaldehyde by peroxidase in the presence of sulfite and oxygen. J Biol Chem. 1969 Aug 25;244(16):4360–4365. [PubMed] [Google Scholar]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES