Skip to main content
PMC home page
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1995 Jun;61(6):2408–2413. doi: 10.1128/aem.61.6.2408-2413.1995

Cloning and sequencing of a laccase gene from the lignin-degrading basidiomycete Pleurotus ostreatus.

P Giardina 1, R Cannio 1, L Martirani 1, L Marzullo 1, G Palmieri 1, G Sannia 1
PMCID: PMC167512  PMID: 7793961

Abstract

The gene (pox1) encoding a phenol oxidase from Pleurotus ostreatus, a lignin-degrading basidiomycete, was cloned and sequenced, and the corresponding pox1 cDNA was also synthesized and sequenced. The isolated gene consists of 2,592 bp, with the coding sequence being interrupted by 19 introns and flanked by an upstream region in which putative CAAT and TATA consensus sequences could be identified at positions -174 and -84, respectively. The isolation of a second cDNA (pox2 cDNA), showing 84% similarity, and of the corresponding truncated genomic clones demonstrated the existence of a multigene family coding for isoforms of laccase in P. ostreatus. PCR amplifications of specific regions on the DNA of isolated monokaryons proved that the two genes are not allelic forms. The POX1 amino acid sequence deduced was compared with those of other known laccases from different fungi.

Full Text

The Full Text of this article is available as a PDF (459.2 KB).

Selected References

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

  1. 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]
  2. Bao W., O'malley D. M., Whetten R., Sederoff R. R. A laccase associated with lignification in loblolly pine xylem. Science. 1993 Apr 30;260(5108):672–674. doi: 10.1126/science.260.5108.672. [DOI] [PubMed] [Google Scholar]
  3. Bollag J. M., Shuttleworth K. L., Anderson D. H. Laccase-mediated detoxification of phenolic compounds. Appl Environ Microbiol. 1988 Dec;54(12):3086–3091. doi: 10.1128/aem.54.12.3086-3091.1988. [DOI] [PMC free article] [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. Cenis J. L. Rapid extraction of fungal DNA for PCR amplification. Nucleic Acids Res. 1992 May 11;20(9):2380–2380. doi: 10.1093/nar/20.9.2380. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Choi G. H., Larson T. G., Nuss D. L. Molecular analysis of the laccase gene from the chestnut blight fungus and selective suppression of its expression in an isogenic hypovirulent strain. Mol Plant Microbe Interact. 1992 Mar-Apr;5(2):119–128. doi: 10.1094/mpmi-5-119. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Frohman M. A., Dush M. K., Martin G. R. Rapid production of full-length cDNAs from rare transcripts: amplification using a single gene-specific oligonucleotide primer. Proc Natl Acad Sci U S A. 1988 Dec;85(23):8998–9002. doi: 10.1073/pnas.85.23.8998. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gavel Y., von Heijne G. Sequence differences between glycosylated and non-glycosylated Asn-X-Thr/Ser acceptor sites: implications for protein engineering. Protein Eng. 1990 Apr;3(5):433–442. doi: 10.1093/protein/3.5.433. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Germann U. A., Lerch K. Isolation and partial nucleotide sequence of the laccase gene from Neurospora crassa: amino acid sequence homology of the protein to human ceruloplasmin. Proc Natl Acad Sci U S A. 1986 Dec;83(23):8854–8858. doi: 10.1073/pnas.83.23.8854. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Hattori M., Sakaki Y. Dideoxy sequencing method using denatured plasmid templates. Anal Biochem. 1986 Feb 1;152(2):232–238. doi: 10.1016/0003-2697(86)90403-3. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. 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]
  15. Lucas M. C., Jacobson J. W., Giles N. H. Characterization and in vitro translation of polyadenylated messenger ribonucleic acid from Neurospora crassa. J Bacteriol. 1977 Jun;130(3):1192–1198. doi: 10.1128/jb.130.3.1192-1198.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Marzullo L., Cannio R., Giardina P., Santini M. T., Sannia G. Veratryl alcohol oxidase from Pleurotus ostreatus participates in lignin biodegradation and prevents polymerization of laccase-oxidized substrates. J Biol Chem. 1995 Feb 24;270(8):3823–3827. doi: 10.1074/jbc.270.8.3823. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Nicole M., Chamberland H., Geiger J. P., Lecours N., Valero J., Rio B., Ouellette G. B. Immunocytochemical localization of laccase L1 in wood decayed by Rigidoporus lignosus. Appl Environ Microbiol. 1992 May;58(5):1727–1739. doi: 10.1128/aem.58.5.1727-1739.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Palmieri G., Giardina P., Marzullo L., Desiderio B., Nitti G., Cannio R., Sannia G. Stability and activity of a phenol oxidase from the ligninolytic fungus Pleurotus ostreatus. Appl Microbiol Biotechnol. 1993 Jul;39(4-5):632–636. doi: 10.1007/BF00205066. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. 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]
  22. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Sannia G., Limongi P., Cocca E., Buonocore F., Nitti G., Giardina P. Purification and characterization of a veratryl alcohol oxidase enzyme from the lignin degrading basidiomycete Pleurotus ostreatus. Biochim Biophys Acta. 1991 Jan 23;1073(1):114–119. doi: 10.1016/0304-4165(91)90190-r. [DOI] [PubMed] [Google Scholar]
  24. Sterjiades R., Dean J. F., Eriksson K. E. Laccase from Sycamore Maple (Acer pseudoplatanus) Polymerizes Monolignols. Plant Physiol. 1992 Jul;99(3):1162–1168. doi: 10.1104/pp.99.3.1162. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. von Heijne G. A new method for predicting signal sequence cleavage sites. Nucleic Acids Res. 1986 Jun 11;14(11):4683–4690. doi: 10.1093/nar/14.11.4683. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES