通过单次单因子试验和正交试验,对Cp-S316菌株抗真菌活性物质的发酵培养基进行了优化,获得适合菌株生长和抗真菌活性物质产生的最佳培养基配方为:土豆100g、乳糖40g、蛋白胨15g、硝酸钠0.5g、硫酸镁2g、水1L。经摇瓶发酵试验,发酵液效价比基础培养基配方提高了325.24%。Cp-S316菌株产生的抗真菌活性物质粗提物对烟草赤星病菌菌丝具有强烈的抑制作用,采用抑制菌丝生长速率法测定了其对烟草赤星病菌的抑制中浓度为EC50=0.248mg·mL^-1;显微镜下观察发现,活性物质能够引起菌丝原生质凝集。
The fermentation medium of Cp-S316 that could produce antifungal active substance was optimized by one-factor-at-a-time and orthogonal design method. The medium components, including different carbon sources, nitrogen sources, C/N ratio and inorganic salt, were considered, and the obtained optimum medium good for the growth of Cp-S316 and production of active substance contained potato 100 g, lactose 40 g, peptone15 g, NaNO3 0.5 g, MgSO4 2 g and H20 1 L. Production of antifungal active substance under optimum medium was 325.24% higher than that under basal medium in shake flask fermentation. The crude antifungal active substance was obtained by the methods of macroporeus resin X-5 adsorption and desorption, evaporation for concentration and lyophilization, and had strong iubibition against Alternaria alternate, with EC50 value being 0.248 mg·mL^-1 determined by means of mycelium growth rate. Observed by microscope, the mycelium protoplast agglomerated under the treatment of antifungal active substance.