Abstract
A purification scheme involving ion exchange chromatography on diethylaminoethyl (DEAE) cellulose, carboxymethyl (CM) cellulose, and Q-Sepharose, followed by fast protein liquid chromatography–gel filtration on Superdex 75, was utilized to isolate a laccase from the fruiting bodies of the mushroom Pleurotus eryngii. The laccase was adsorbed on DEAE-cellulose and Q-Sepharose and unadsorbed on CM-cellulose. The molecular mass of the laccase in sodium dodecyl sulfate–polyacrylamide gel electrophoresis was 34 kDa. The laccase activity was maximal at 70°C and remained high at 80°C. High activity of the laccase was maintained at ph 3 to 5. The activity underwent an abrupt decline at pH 6 and 7 and was indiscernible at ph 8 and 9. The laccase exhibited an inhibitory activity toward HIV-1 reverse transcriptase with an IC50 of 2.2 μM. Its N-terminal sequence was dissimilar to those of laccase isoenzymes previously isolated from cultured mycelia of the same species.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Cambria M, Cambria A, Ragusa S, Rizzarelli E (2000) Production, purification and properties of an extracellular laccase from Rigidoporus lignosus. Protein Expr Purif 18:141–147
Dedeyan B, Klonowska A, Tagger S, Tron T, Iacazio G, Gil G, Le Petit J (2000) Biochemical and molecular characterization of a laccase from Marasmius guercophilus. Appl Environ Microbiol 66:925–929
Endo K, Hosono K, Beppu T, Udea K (2002) A novel extracytoplasmic phenol oxidase of Streptomyces: its possible involvement in the onset of morphogenesis. Microbiology 148:1767–1776
Endo K, Hayashi Y, Hibi T, Hosono K, Beppu T, Ueda K (2003) Enzymological characterization of EpoA, a laccase-like phenol oxidase produced by Streptomyces griseus. J Biochem 133:671–677
Garzillo AM, Colao MC, Buonocore V, Oliva R, Falcigno L, Saviano M, Santoro AM, Zappala R, Ronomo RP, Bianco C, Giardina P, Palmieri G, Sannia G (2001) Structural and kinetic characterization of native laccases from Pleurotus ostreatus, Rigidoporus lignosus and Trametes trogii. J Protein Chem 20:191–201
Laemmli UK, Favre M (1973) Gel electrophoresis of proteins. J Mol Biol 80:575–599
Machczynski MC, Vjgenboom E, Samyn B, Canters GW (2004) Characterization of SLAC: a small laccase from Streptomyces coelicolor with unprecedented activity. Protein Sci 13:2388–2397
Martirani L, Giardina P, Marzullo L, Sannia G (1996) Reduction of phenol content and toxicity in olive oil mill waste waters with the ligninolytic fungus Pleurotus ostreatus. Water Res 30:1914–1918
Monoz C, Guillen F, Martinozv, Martinez MJ (1997) Laccase isoenzymes of Pleurotus eryngii: characterization, catalytic properties, and participation in activation of molecular oxygen and Mn2+ oxidation. Appl Environ Microbiol 63:2166–2174
Nakamura K, Kawabata T, Yura K, Go N (2003) Novel types of two-domain multi-copper oxidases: possible missing links in the evolution. FEBS Lett 553:239–244
Ng TB, Huang B, Fong WP, Yeung HW (1997) Anti-HIV natural products with special emphasis on HIV-1 reverse transcriptase inhibitors. Life Sci 61:933–949
Nomura N, Inokuchi N, Kobayashi H, Koyama T, Iwama M, Ohgi K, Irie M (1994) Purification and primary structure of a new guanylic acid-specific ribonuclease from Pleurotus ostreatus. J Biochem (Tokyo) 116:26–33
Palmieri G, Giardina P, Desiderio B, Marzullo L, Giamberini M, Sannia G (1993) A new enzyme immobilization procedure using copper alginate gel: application to a fungal phenol oxidase. Enzyme Microb Technol 16:151–158
Perry CR, Matcham SE, Ward DA, Thurston CF (1993) The structure of laccase protein and its synthesis by the commercial mushroom Agaricus bisporus. J Gen Microbiol 139:171–179
Reid ID, Paice MG (1994) Biological bleaching of kraft pulps by white-rot fungi and their enzymes. FEMS Microbiol Rev 13:369–376
She QB, Ng TB, Liu WK (1998) A novel lectin with potent immunomodulatory activity isolated from both fruiting bodies and cultured mycelia of the edible mushroom Volvariella volvacea. Biochem Biophys Res Commun 247:106–111
Shin KS, Lee YJ (2000) Purification and characterization of a new member of the laccase family from the white-rot basidiomycete Coriolus hirsutus. Arch Biochem Biophys 384:109–115
Wang HX, Ng TB (2001a) Pleureryn, a novel protease from fresh fruiting bodies of the edible mushroom Pleurotus eryngii. Biochem Biophys Res Commun 289:750–755
Wang HX, Ng TB (2001b) Examination of lectins, polysaccharopeptide, polysaccharide, alkaloid, coumarin and trypsin inhibitors for inhibitory activity against human immunodeficiency virus transcriptase and glycohydrolase. Planta Med 67:669–672
Wang HX, Ng TB (2004a) A homodimeric laccase with unique characteristics from the yellow mushroom Cantharellus cibarius. Biochem Biophys Res Commun 313:37–41
Wang HX, Ng TB (2004b) Purification of a novel low-molecular-mass laccase with HIV-1 reverse transcriptase inhibitory activity from the mushroom Tricholoma giganteum. Biochem Biophys Res Commun 315:450–454
Wang HX, Ng TB, Liu WK, Ooi VEC, Chang ST (1995) Isolation and characterization of two distinct lectins with antiproliferative activity from the cultured mycelium of the edible mushroom Tricholoma mongolicum. Int J Pept Protein Res 46:508–513
Wang HX, Ng TB, Ooi VEC, Liu WK, Chang ST (1996) A polysaccharide–peptide complex from cultured mycelia of the mushroom Tricholoma mongolicum with immunoenhancing and antitumor activities. Biochem Cell Biol 74:95–100
Wang HX, Ng TB, Ooi VEC (1998) Lectin activity in fruiting bodies of the edible mushroom Tricholoma mongolicum. Biochem Mol Biol Int 44:135–141
Wang HX, Gao J, Ng TB (2000) A new lectin with highly potent antihepatoma and antisarcoma activities from the oyster mushroom Pleurotus ostreatus. Biochem Biophys Res Commun 275:810–816
Yoshioka Y, Tabeta R, Saito H, Uehara N, Fukuoka F (1985) Antitumor polysaccharides from Pleurotus ostreatus (Fr.) Ouel: isolation and structure of a beta-glucan. Carbohydr Res 140:93–100
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Wang, H.X., Ng, T.B. Purification of a laccase from fruiting bodies of the mushroom Pleurotus eryngii . Appl Microbiol Biotechnol 69, 521–525 (2006). https://doi.org/10.1007/s00253-005-0086-7
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00253-005-0086-7