摘要
文章采用海藻酸钠与明胶为载体,对磁性Fe 3 O 4进行包埋,使其形成磁性复合载体凝胶微球,经戊二醛交联后,固定生姜蛋白酶,得到磁性复合载体固定生姜蛋白酶;在预实验的基础上,确定影响制备固定化生姜蛋白酶的主要因素;并研究在最优条件下,固定化生姜蛋白酶与其游离酶的酶学性质。结果表明:磁性复合载体固定化生姜蛋白酶的酸碱稳定性范围为pH 3.0~7.0,在20~60℃内具有较好的抗热性,其最适pH为4.0,温度为50℃,表观米氏常数Kmapp为3.162 mg/mL;在此条件下固定60 min,制备的固定化生姜蛋白酶剩余酶活力可达78.69%。重复使用10次后,酶活力剩余53%;在4℃下储存10 d,酶活力仍保留50.66%。说明在最优工艺条件下所制备的固定化酶机械强度大,弹性好,操作稳定性强,酶活力回收率高。
In this paper,sodium alginate and gelatin are used as the carriers to embed magnetic Fe 3 O 4 to form magnetic composite carrier gel microspheres.After cross-linking with glutaraldehyde,ginger protease is immobilized to obtain magnetic composite carrier immobilized ginger protease.On the basis of preliminary experiments,the main factors affecting the preparation of immobilized ginger protease are determined.The enzymatic properties of immobilized ginger protease and its free enzyme are studied under the optimal conditions.The results show that the acid-base stability of immobilized ginger protease with magnetic composite carrier ranges from pH 3.0 to 7.0,and it has good heat resistance at 20~60℃.The optimum pH is 4.0 and the temperature is 50℃.The Kmapp constant is 3.162 mg/mL;under these conditions for 60 min,the residual enzyme activity of the prepared immobilized ginger protease can reach 78.69%.After repeated use for 10 times,the enzyme activity remains 53%;when stored at 4℃for 10 days,the enzyme activity remains 50.66%.The results show that the immobilized enzyme prepared under the optimal process conditions has high mechanical strength,good elasticity,strong operation stability and high recovery rate of enzyme activity.
作者
罗璇
陈常缘
王秀原
杜婧加
刘婧雨
吴蒙
LUO Xuan;CHEN Chang-yuan;WANG Xiu-yuan;DU Jing-jia;LIU Jing-yu;WU Meng(School of Food and Biotechnology,Wuhan Institute of Design and Sciences,Wuhan 430205,China)
出处
《中国调味品》
CAS
北大核心
2020年第3期57-61,共5页
China Condiment
基金
2019年武汉设计工程学院科研项目(K201911)
2018年国家级大学生科技创新项目(201814035006)
关键词
磁性复合载体
酶学性质
固定化
生姜蛋白酶
magnetic composite carrier
enzymatic property
immobilization
ginger protease
