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Volume 149, Issue 9

Important role of fungal intracellular laccase for melanin synthesis: purification and characterization of an intracellular laccase from fruit bodies Free

Abstract

A laccase (EC 1.10.3.2) was isolated from the fully browned gills of fruit bodies. The enzyme was purified to a homogeneous preparation using hydrophobic, cation-exchange and size-exclusion chromatography. SDS-PAGE analysis showed the purified laccase, Lcc 2, to be a monomeric protein of 58·0 kDa. The enzyme had an isoelectric point of around pH 6·9. The optimum pH for enzyme activity was around 3·0 against 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)diammonium salt (ABTS), and it was most active at 40 °C and stable up to 50 °C. The enzyme contained 8·6 % carbohydrate and some copper atoms. The enzyme oxidized ABTS, -phenylenediamine, pyrogallol, guaiacol, 2,6-dimethoxyphenol, catechol and ferulic acid, but not veratryl alcohol and tyrosine. -(3,4-Dihydroxyphenyl)alanine (-DOPA), which was not oxidized by a laccase previously reported from the culture filtrate of , was also oxidized by Lcc 2, and the oxidative product of -dopa was identified as -DOPA quinone by HPLC analysis. Lcc 2 was able to oxidize phenolic compounds extracted from fresh gills to brown-coloured products, suggesting a role for laccase in melanin synthesis in this strain.

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Keyword(s): ABTS, 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)diammonium salt , DHN, dihydroxynaphthalene , GDHB, γ-glutaminyl-3,4-dihydroxybenzene , GHB, γ-glutaminyl-4-hydroxybenzene , l-DOPA, β-(3,4-dihydroxyphenyl)alanine , Lcc, laccase , PB, 10 mM sodium phosphate buffer and Tyr, tyrosinase
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2003-09-01
2024-04-25
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Important role of fungal intracellular laccase for melanin synthesis: purification and characterization of an intracellular laccase from Lentinula edodes fruit bodies
Microbiology 149, 2455 (2003); https://doi.org/10.1099/mic.0.26414-0
/content/journal/micro/10.1099/mic.0.26414-0
/content/journal/micro/10.1099/mic.0.26414-0
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