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Purification and characterization of the extracellular laccase produced by Trametes polyzona WR710–1 under solid‐state fermentation
Author(s) -
Chairin Thanunchanok,
Nitherat Thitinard,
Watanabe Akira,
Asada Yasuhiko,
Khagnuch Chartchai,
Lumyong Saisamorn
Publication year - 2014
Publication title -
journal of basic microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.58
H-Index - 54
eISSN - 1521-4028
pISSN - 0233-111X
DOI - 10.1002/jobm.201200456
Subject(s) - laccase , abts , guaiacol , catechol , solid state fermentation , chemistry , molecular mass , fermentation , nuclear chemistry , orange (colour) , enzyme , chromatography , substrate (aquarium) , biochemistry , food science , dpph , biology , antioxidant , ecology
Laccase from Trametes polyzona WR710–1 was produced under solid‐state fermentation using the peel from the Tangerine orange ( Citrus reticulata Blanco) as substrate, and purified to homogeneity. This laccase was found to be a monomeric protein with a molecular mass of about 71 kDa estimated by SDS–PAGE. The optimum pH was 2.0 for ABTS, 4.0 for L ‐DOPA, guaiacol, and catechol, and 5.0 for 2,6‐DMP. The K m value of the enzyme for the substrate ABTS was 0.15 mM, its corresponding V max value was 1.84 mM min −1 , and the k cat / K m value was about 3960 s −1 mM −1 . The enzyme activity was stable between pH 6.0 and 8.0, at temperatures of up to 40 °C. The laccase was inhibited by more than 50% in the presence of 20 mM NaCl, by 95% at 5 mM of Fe 2+ , and it was completely inhibited by 0.1 mM NaN 3 . The N‐terminal amino acid sequence of this laccase is AVTPVADLQISNAGISPDTF, which is highly similar to those of laccases from other white‐rot basidiomycetes.