以碱性蛋白酶生产菌克劳氏芽孢杆菌(Bacillus clausii)基因组DNA为模板PCR扩增获得尿酸氧化酶基因(BcU),插入原核表达载体pET28α中,构建表达载体pET-BcU,并转化大肠杆菌BL21(DE3)获得重组大肠杆菌BL21(DE3)/pET-BcU.经IPTG诱导,重组菌BL21(DE3)/pET-BcU表达出有活性的尿酸氧化酶,含空质粒的重组菌在同样条件下没有酶活.酶学性质分析显示,重组酶最适pH值为9.0,在pH值9.0~11范围内酶活几乎不变,是一种高碱性尿酸氧化酶.%The BcU gene encoding urate oxidase was amplified by PCR with genome DNA of Bacillus clausii as template. The gene BcU was cloned into pET28α resulting in the recombinant plasmid pET-BcU. The recombiant plasmid was transformed into Escherichia coli BL21(DE3). Induced with IPTG, the recombinant strain BL21 ( DE3 )/pET-BcU expressed active urate oxidase, while the control BL21 ( DE3 )/pET did not. The optimum pH for recombinant enzyme was 9. 0 and the enzymatic activity showed almost no difference between pH 9. 0 ~ 11. The recombinant enzyme is a kind of high alkaline urate oxidase.
展开▼
