Skip to main content
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1995 Mar;61(3):907–912. doi: 10.1128/aem.61.3.907-912.1995

Production and Characterization of Laccase from Botrytis cinerea 61-34

D Slomczynski, J P Nakas, S W Tanenbaum
PMCID: PMC1388373  PMID: 16534974

Abstract

An isolate of Botrytis cinerea (strain 61-34) constitutively expresses substantial amounts of extracellular laccase on a defined growth medium. The enzyme has been purified to homogeneity by a facile operational sequence, the last stage of which involves hydrophobic interaction chromatography. By these means, over 80 mg of laccase liter(sup-1) can be obtained from aerated fermentor reaction broths. The enzyme, with an estimated M(infr) of 74,000 and pI of 4.0, is a monomeric glycoprotein containing 49% carbohydrate predominantly as hexose. With 2,6-dimethoxyphenol, it exhibits a pH optimum of 3.5 and a temperature optimum of 60(deg)C, and its K(infm) is 100 (mu)M. The purified enzyme with this substrate has a specific activity of 9.1 mkat mg of protein(sup-1). Taken together with a broad substrate range and its stability in 4% sodium dodecyl sulfate or 2 M urea solutions, several biotechnology transfers are suggested.

Full Text

The Full Text of this article is available as a PDF (233.8 KB).

Selected References

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

  1. Bollag J. M., Leonowicz A. Comparative studies of extracellular fungal laccases. Appl Environ Microbiol. 1984 Oct;48(4):849–854. doi: 10.1128/aem.48.4.849-854.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Canters G. W., Gilardi G. Engineering type 1 copper sites in proteins. FEBS Lett. 1993 Jun 28;325(1-2):39–48. doi: 10.1016/0014-5793(93)81410-2. [DOI] [PubMed] [Google Scholar]
  3. Coll P. M., Fernández-Abalos J. M., Villanueva J. R., Santamaría R., Pérez P. Purification and characterization of a phenoloxidase (laccase) from the lignin-degrading basidiomycete PM1 (CECT 2971). Appl Environ Microbiol. 1993 Aug;59(8):2607–2613. doi: 10.1128/aem.59.8.2607-2613.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  5. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  6. Leach B. S., Collawn J. F., Jr, Fish W. W. Behavior of glycopolypeptides with empirical molecular weight estimation methods. 1. In sodium dodecyl sulfate. Biochemistry. 1980 Dec 9;19(25):5734–5741. doi: 10.1021/bi00566a011. [DOI] [PubMed] [Google Scholar]
  7. Messerschmidt A. Structural studies on copper-containing plant oxidases. Biochem Soc Trans. 1992 May;20(2):364–368. doi: 10.1042/bst0200364. [DOI] [PubMed] [Google Scholar]
  8. Patel T., Bruce J., Merry A., Bigge C., Wormald M., Jaques A., Parekh R. Use of hydrazine to release in intact and unreduced form both N- and O-linked oligosaccharides from glycoproteins. Biochemistry. 1993 Jan 19;32(2):679–693. doi: 10.1021/bi00053a037. [DOI] [PubMed] [Google Scholar]
  9. Rigling D., Van Alfen N. K. Extra- and Intracellular Laccases of the Chestnut Blight Fungus, Cryphonectria parasitica. Appl Environ Microbiol. 1993 Nov;59(11):3634–3639. doi: 10.1128/aem.59.11.3634-3639.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Sariaslani F. S. Microbial enzymes for oxidation of organic molecules. Crit Rev Biotechnol. 1989;9(3):171–257. doi: 10.3109/07388558909036736. [DOI] [PubMed] [Google Scholar]
  12. Stahmann K. P., Pielken P., Schimz K. L., Sahm H. Degradation of Extracellular beta-(1,3)(1,6)-d-Glucan by Botrytis cinerea. Appl Environ Microbiol. 1992 Oct;58(10):3347–3354. doi: 10.1128/aem.58.10.3347-3354.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Tamaru H., Inoue H. Isolation and characterization of a laccase-derepressed mutant of Neurospora crassa. J Bacteriol. 1989 Nov;171(11):6288–6293. doi: 10.1128/jb.171.11.6288-6293.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES