米曲霉腺苷酸脱氨酶的产酶条件

Conditions of Preparing AMP Deaminase from Aspergillus oryzae

通过实验获得了米曲霉 3.80 0固态发酵产腺苷酸 (AMP)脱氨酶的适宜培养基 ,麸皮为主原料 ,成分质量分数为蔗糖 2 % ,鱼粉 2 % ,(NH4 ) 2 SO4 0 .1% ,吐温 80 0 .1% ,含水量 5 0 % .最佳的培养条件为 :2 5 0mL的三角瓶装 2 0 g培养基 ,在 2 8～ 30℃培养 6 0h .发酵结束称取一定量的麸曲加 10倍质量的蒸馏水 ,调节 pH 6 .0 ,在 35℃条件振荡水浴 2h .酶的浸提液经盐析、透析、DEAE柱层析分离后 ,可收集到较单一的活性峰酶 .

Aspergillus oryzae 3.800 was selected for producing AMP deaminase, and optimal composition of culture medium for enzyme secretion was: wheat bran as main material, sucrose 2%, fish meal 2%, (NH 4) 2SO 4 0.1%, polysorbatum 80 0.1%, water hold 50%. The best cultivating conditions were as follows: 20 g medium in 250 mL flask, the fungus was incubated at 28～30 ℃ for 60 h. The enzyme was extracted by adding water to moldy bran after fermentation to a ratio of 1∶10, pH 6.0, and shaking for 2 hours under 35 ℃. AMP deaminase was preliminary purified through precipitation by (NH 4) 2SO 4, dialysis and DEAE Spherical Agarose chromatography, the single enzyme active peak was acquired.