Abstract: The experimental study the optimal culture conditions for laccase production by Melanoporia castanea in shake-flask cultivation And use Minitab15 software, Plackett-Burman PB experimental design and response surface analysis optimize Melanoporia castanea (Imazeki) T. Hatt. & Ryvarden laccase fermentation conditions.The composition of fermentation medium :Corn stalks 9.976g/L, Soybean Meal 9g / L, ZnSO4•7H2O 0.3μmol / L, p-phenylenediamine 0.5mmol / L. Optimal culture conditions: temperature 28 ℃, speed 180r/min, PH 5.913, Liquid volume 60ml/250ml, inoculum 10%. Under the above cultured conditions, laccase activity can be achieved 431U / ml.