Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Purification and Characterization of a Laccase from the Edible Wild Mushroom Tricholoma mongolicum

  • Li, Miao (State Key Laboratory for Agrobiotechnology and Department of Microbiology, China Agricultural University) ;
  • Zhang, Guoqing (State Key Laboratory for Agrobiotechnology and Department of Microbiology, China Agricultural University) ;
  • Wang, Hexiang (State Key Laboratory for Agrobiotechnology and Department of Microbiology, China Agricultural University) ;
  • Ng, Tzibun (School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong)
  • Received : 2009.12.24
  • Accepted : 2010.05.03
  • Published : 2010.07.28
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Abstract

A novel laccase from Tricholoma mongolicum was purified by using a procedure that entailed ion-exchange chromatographies on DEAE-cellulose, CM-cellulose, and Q-Sepharose, and FPLC-gel filtration on Superdex 75. The purified enzyme was obtained with a specific activity of 1,480 U/mg-protein and a final yield of 15%. It was found to be a monomeric protein with a molecular mass of 66 kDa as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Its N-terminal amino acid sequence was GIGPVADLYVGNRIL, similar to some but also different to other mushroom laccases. The optimum pH and temperature for the purified enzyme were pH 2 to pH 3 and $30^{\circ}C$, respectively. It displayed a low $K_m$ toward 2,7-azinobis (3-ethylbenzothiazolone-6-sulfonic acid) diammonium salt (ABTS) and high $k_{cat}/K_m$ values. The purified laccase oxidized a wide range of lignin-related phenols, but exerted maximal activity on ABTS. It was significantly inhibited by $Hg^{2+}$ ions, and remarkably stimulated by $Cu^{2+}$ ions. It inhibited HIV-1 reverse transcriptase and proliferation of hepatoma HepG2 cells and breast cancer MCF7 cells with an $IC_{50}$ of 0.65 ${\mu}M$, 1.4 ${\mu}M$, and 4.2 ${\mu}M$, respectively, indicating that it is also an antipathogenic protein.

Keywords

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