摘要
通过对米曲氨基酰化酶硫酸铵分级沉淀、疏水层析分离、SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)实验获得较高纯化倍数和较高活力的酶,并对酶保存稳定性进行研究。经分离纯化实验后,米曲氨基酰化酶的比活力由3.818 U/mg提高到106.738 U/mg,纯化倍数达28倍,活力回收率为22%。保存该酶的最适pH值为6.5～7.0,Co2+对酶有轻微的激活作用,Fe2+和CH3COO-对酶有明显地抑制作用,蛋白水解酶抑制剂1,4-二硫苏糖醇对酶的保存稳定性有较高提升。自制丙酮冻干粉保存稳定性良好,保存半年其酶活力仍能维持原酶活力的80%以上。
Aminoacylase （EC3.5.1.14） from Asperigillus oryzae No.3042 was isolated and purified by the combination of ammonium sulfate fractionation and hydrophobic chromatography. The optimum techniques of separation and conservancy stability were studied, in order to further research enzyme characteristic and technique of resolution. The specific activity of aminoacylase was 106.738 U/mg, the purification ratio and recovery was 28 and 22%. The optimal pH of aminoacylase was 6.5-7.0. The ions in buffer lowered the activity of aminoacylase, but the Co^2＋ in low concentration activated the aminoacylase. The stability of aminoacylase was improved by adding 1,4-Dithiothreitol （DTT）. When freeze-drying powder of aminoacylase was laid up for half year, the activity of aminoacylase was 80% of original one.
出处
《食品科技》
CAS
北大核心
2008年第4期12-15,共4页
Food Science and Technology
基金
西南交通大学青年教师科研启动项目
