Skip to main content
PMC home page
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1996 Oct;62(10):3739–3744. doi: 10.1128/aem.62.10.3739-3744.1996

Isolation of laccase gene-specific sequences from white rot and brown rot fungi by PCR.

T M D'Souza 1, K Boominathan 1, C A Reddy 1
PMCID: PMC168181  PMID: 8837429

Abstract

Degenerate primers corresponding to the consensus sequences of the copper-binding regions in the N-terminal domains of known basidiomycete laccases were used to isolate laccase gene-specific sequences from strains representing nine genera of wood rot fungi. All except three gave the expected PCR product of about 200 bp. Computer searches of the databases identified the sequence of each of the PCR products analyzed as a laccase gene sequence, suggesting the specificity of the primers. PCR products of the white rot fungi Ganoderma lucidum, Phlebia brevispora, and Trametes versicolor showed 65 to 74% nucleotide sequence similarity to each other; the similarity in deduced amino acid sequences was 83 to 91%. The PCR products of Lentinula edodes and Lentinus tigrinus, on the other hand, showed relatively low nucleotide and amino acid similarities (58 to 64 and 62 to 81%, respectively); however, these similarities were still much higher than when compared with the corresponding regions in the laccases of the ascomycete fungi Aspergillus nidulans and Neurospora crassa. A few of the white rot fungi, as well as Gloeophyllum trabeum, a brown rot fungus, gave a 144-bp PCR fragment which had a nucleotide sequence similarity of 60 to 71%. Demonstration of laccase activity in G. trabeum and several other brown rot fungi was of particular interest because these organisms were not previously shown to produce laccases.

Full Text

The Full Text of this article is available as a PDF (719.3 KB).

Selected References

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

  1. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
  2. Aramayo R., Timberlake W. E. Sequence and molecular structure of the Aspergillus nidulans yA (laccase I) gene. Nucleic Acids Res. 1990 Jun 11;18(11):3415–3415. doi: 10.1093/nar/18.11.3415. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Ballance D. J. Sequences important for gene expression in filamentous fungi. Yeast. 1986 Dec;2(4):229–236. doi: 10.1002/yea.320020404. [DOI] [PubMed] [Google Scholar]
  4. Bourbonnais R., Paice M. G. Oxidation of non-phenolic substrates. An expanded role for laccase in lignin biodegradation. FEBS Lett. 1990 Jul 2;267(1):99–102. doi: 10.1016/0014-5793(90)80298-w. [DOI] [PubMed] [Google Scholar]
  5. Coll P. M., Tabernero C., Santamaría R., Pérez P. Characterization and structural analysis of the laccase I gene from the newly isolated ligninolytic basidiomycete PM1 (CECT 2971). Appl Environ Microbiol. 1993 Dec;59(12):4129–4135. doi: 10.1128/aem.59.12.4129-4135.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dass S. B., Reddy C. A. Characterization of extracellular peroxidases produced by acetate-buffered cultures of the lignin-degrading basidiomycete Phanerochaete chrysosporium. FEMS Microbiol Lett. 1990 Jun 1;57(3):221–224. doi: 10.1111/j.1574-6968.1990.tb04233.x. [DOI] [PubMed] [Google Scholar]
  7. Fukushima Y., Kirk T. K. Laccase component of the Ceriporiopsis subvermispora lignin-degrading system. Appl Environ Microbiol. 1995 Mar;61(3):872–876. doi: 10.1128/aem.61.3.872-876.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Germann U. A., Müller G., Hunziker P. E., Lerch K. Characterization of two allelic forms of Neurospora crassa laccase. Amino- and carboxyl-terminal processing of a precursor. J Biol Chem. 1988 Jan 15;263(2):885–896. [PubMed] [Google Scholar]
  9. Giardina P., Cannio R., Martirani L., Marzullo L., Palmieri G., Sannia G. Cloning and sequencing of a laccase gene from the lignin-degrading basidiomycete Pleurotus ostreatus. Appl Environ Microbiol. 1995 Jun;61(6):2408–2413. doi: 10.1128/aem.61.6.2408-2413.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kirk T. K., Farrell R. L. Enzymatic "combustion": the microbial degradation of lignin. Annu Rev Microbiol. 1987;41:465–505. doi: 10.1146/annurev.mi.41.100187.002341. [DOI] [PubMed] [Google Scholar]
  11. Kojima Y., Tsukuda Y., Kawai Y., Tsukamoto A., Sugiura J., Sakaino M., Kita Y. Cloning, sequence analysis, and expression of ligninolytic phenoloxidase genes of the white-rot basidiomycete Coriolus hirsutus. J Biol Chem. 1990 Sep 5;265(25):15224–15230. [PubMed] [Google Scholar]
  12. Marchuk D., Drumm M., Saulino A., Collins F. S. Construction of T-vectors, a rapid and general system for direct cloning of unmodified PCR products. Nucleic Acids Res. 1991 Mar 11;19(5):1154–1154. doi: 10.1093/nar/19.5.1154. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Messerschmidt A., Huber R. The blue oxidases, ascorbate oxidase, laccase and ceruloplasmin. Modelling and structural relationships. Eur J Biochem. 1990 Jan 26;187(2):341–352. doi: 10.1111/j.1432-1033.1990.tb15311.x. [DOI] [PubMed] [Google Scholar]
  14. Niku-Paavola M. L., Karhunen E., Salola P., Raunio V. Ligninolytic enzymes of the white-rot fungus Phlebia radiata. Biochem J. 1988 Sep 15;254(3):877–883. doi: 10.1042/bj2540877. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Orth A. B., Royse D. J., Tien M. Ubiquity of lignin-degrading peroxidases among various wood-degrading fungi. Appl Environ Microbiol. 1993 Dec;59(12):4017–4023. doi: 10.1128/aem.59.12.4017-4023.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Perry C. R., Smith M., Britnell C. H., Wood D. A., Thurston C. F. Identification of two laccase genes in the cultivated mushroom Agaricus bisporus. J Gen Microbiol. 1993 Jun;139(Pt 6):1209–1218. doi: 10.1099/00221287-139-6-1209. [DOI] [PubMed] [Google Scholar]
  17. Rao T. R., Reddy C. A. DNA sequences from a ligninolytic filamentous fungus Phanerochaete chrysosporium capable of autonomous replication in yeast. Biochem Biophys Res Commun. 1984 Feb 14;118(3):821–827. doi: 10.1016/0006-291x(84)91468-2. [DOI] [PubMed] [Google Scholar]
  18. Reddy C. A., D'Souza T. M. Physiology and molecular biology of the lignin peroxidases of Phanerochaete chrysosporium. FEMS Microbiol Rev. 1994 Mar;13(2-3):137–152. doi: 10.1111/j.1574-6976.1994.tb00040.x. [DOI] [PubMed] [Google Scholar]
  19. Saloheimo M., Niku-Paavola M. L., Knowles J. K. Isolation and structural analysis of the laccase gene from the lignin-degrading fungus Phlebia radiata. J Gen Microbiol. 1991 Jul;137(7):1537–1544. doi: 10.1099/00221287-137-7-1537. [DOI] [PubMed] [Google Scholar]
  20. Yaver D. S., Xu F., Golightly E. J., Brown K. M., Brown S. H., Rey M. W., Schneider P., Halkier T., Mondorf K., Dalboge H. Purification, characterization, molecular cloning, and expression of two laccase genes from the white rot basidiomycete Trametes villosa. Appl Environ Microbiol. 1996 Mar;62(3):834–841. doi: 10.1128/aem.62.3.834-841.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES