Figure 3
Expression, purification and zymogram analysis of purified laccase. (A) PAGE separation of proteins from culture supernatant of P. pastoris pPICZαB lcc-5 and confirmation of laccase secretion by zymogram analysis using guaiacol as substrate. Lane 1, Pre-stained molecular weight markers. Lanes 3,4: 10, 20 μl respectively of the culture filtrate. Lane 5, 20 μl of the culture filtrate in loading buffer not containing SDS. Lane 6: 20 μl of the culture filtrate in loading buffer not containing β–mercaptoethanol. (B) SDS-PAGE analysis of the purified laccase. Lane 1, molecular weight markers. Lane 2, purified nLac (~5 μg). Lane 3, purified rLac from P. pastoris (~7 μg). (C) PAGE-Zymogram analysis using ABTS as a substrate. Lane 1, Pre-stained molecular weight markers. Lane 2, purified nLac. Lane 3, purified rLac.