研究了重组表达人源过氧化氢酶酵母菌株G13的发酵工艺和纯化方法.对接种量、生长期培养基、生长时间、诱导时间等因素进行了优化,并确立了10 L发酵罐的发酵条件.结果表明,菌体最高密度达到了0.15 g/mL,比原重组菌株S4提高了1倍,人源过氧化氢酶表达量也由原来的平均600 U/mL提高到了1500 U/mL.同时建立了二步法纯化该酶工艺,获得了电泳纯的人源过氧化氢酶.所得到的重组蛋白经SDS-PAGE和Western-blot鉴定,证实产物为人源过氧化氢酶.%The ferm entation conditios such as inoculum size , growth medjum , growth phase and induced time by a new recom binantyeast strain G 13 were investigated and its new purification method was prsented . Under the optim ized conditions , the ferm entation in 10 L fenn entor was performed . The results showed that the expression level of rhcatalase was increased from 600 U /m L to 1500 U /m L . The purified rhcC atalase was obtained through an ion-exchange :resin and verified by Westem-blot. Its specfic activity was equal to the level of com m erial products .
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