Patent ID: 8759057

Claim:
A process for purifying laccase from an ectomycorrhizal fruiting body, the process comprising: (A) homogenizing the fruiting body in a buffer of about pH 7 to obtain a slurry; (B) sonicating the slurry to obtain a sonicated slurry; (C) centrifuging the sonicated slurry at a relative centrifugal force of about 15,000 to about 20,000 and collecting a resulting first supernatant; (D) sonicating the first supernatant to obtain a sonicated first supernatant; (E) ultracentrifuging the sonicated first supernatant such that a resulting second supernatant separates into top and bottom supernatant layers, and collecting the bottom supernatant layer; (F) filtering the bottom supernatant layer through a bacteria-retaining filter and collecting the filtrate; (G) subjecting the filtrate to DEAE cellulose chromatography and eluting retained proteins with a first sodium chloride gradient; (H) removing the sodium chloride from the eluted, retained proteins to obtain desalted proteins, subjecting the desalted proteins to ion exchange chromatography, desorbing adsorbed proteins with a second sodium chloride gradient, and collecting the first desorbed peak; and (I) fractionating the first desorbed peak into fractions by gel filtration.