Biochemical and Enzymatic Properties of Novel Laccase from Fomitopsis pinicola Mycelia

A type of ligninolytic enzyme was produced in the culture supernatant of Fomitopsis pinicola mycelia. The laccase was purified by ion-exchange chromatography on DEAE-Sepharose FF, gel filtration chromatography on Superdex 200 pg, and HIC chromatography on Phenyl FF from the culture supernatant of F. pinicola. The molecular weight of the enzyme was 92 kDa as determined by SDS-PAGE and gel filtration chromatography. The isoelectric point of the enyzme was 3.2 by IEF system. The optimum temperature and pH for the enzyme activity were 80 °C and 3.0, respectively. The enzyme was identified a glycoprotein. As time goes by, the enzyme degraded some dyes, methyl green, remasol brilliant blue R, methylene blue, malachite green, basic red, remasol brilliant violet 5R, janus green, basic blue, bromocresol green and neutral red. The enzyme showed the highest specific activity for 2,2-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid)(ABTS) as a substrate. The Km value of the enzyme for substrate ABTS is 0.28 mM and Vmax value is 4.5 U/ml. The first 15 amino acids of the N-terminal sequence of laccase from F. pinicola DTHKAEIACRFKDLG.