米曲霉氨基酰化酶纯化方法的研究

Study on purification method of aminoacylase of Aspergillus oryzae

比较了盐析法和有机溶剂沉淀法对于米曲霉氨基酰化酶的纯化效果。分别使用硫酸铵、无水乙醇和丙酮对米曲霉氨基酰化酶进行纯化,结果发现:有机溶剂沉淀法的纯化效果优于盐析法。采用丙酮二次沉淀法纯化的氨基酰化酶比酶活为93.20U/mg,纯化倍数为3.26;通过乙醇二次沉淀,得到的酶液比酶活为93.22U/mg,纯化倍数为3.10,而采用盐析法纯化的酶液的比酶活为56.01U/mg,纯化倍数为2.06。从酶的纯化效果方面考虑,丙酮沉淀法最佳,但从工业生产的可行性方面考虑,乙醇沉淀法较好。

The purification of aminoacylase of Aspergillus oryzae was stud- ied using salting out and organic solvent. （NH4）2SO4, ethanol and acetone were used. The result showed that purification by organic solvent was better than that of salting out. The specific activity of aminoacylase purified by acetone was 93.20 U/mg and purification ratio 3.26. The specific activity of aminoacylase purified by ethanol was 93.22 U/mg and purification ratio 3. 10. The specific activity of aminoacylase purified by （ NH4 ） 2SO4 fractionation 56.01 U/mg and purification ratio 2.06. Considered from the purification effect aspect, the acetone fractionation method was the best, but considered from the industrial production feasibility aspect, the ethanol fractionation method was better.