重组甘蔗ACC氧化酶cDNA原核表达的纯化复性研究

Purification and renaturation of recombinant sugarcane ACC oxidase expressed in Escherichia coli

重组甘蔗ACC氧化酶基因在大肠杆菌中表达,其产物以不溶性包涵体存在。用N i2+-NTA亲和层析柱对其进行纯化,结果显示,在层析柱上直接复性及纯化的方法比在变性条件下N i2+-NTA纯化然后稀释透析进行复性的方法简捷,效果好,获得的目的蛋白质纯度大于98%,活性为132.58 nm o l C2H4/(m g.h)。

Recombinant sugarcane ACC oxidase gene was expressed in Escherichia coli BL21 （DE3） plysS, and the product was insoluble inclusion bodies. The recombinant proteins were purified with Ni^2＋-NTA column affinity chromatography. The results showed that the approach for one-step renaturation and purification via Ni^2＋-NTA column was simpler but better than that for renaturation via dilution and dialysis the denatured protein that had been purified with Ni^2＋-NTA. The purity of the target protein obtained from one-step renaturation and purification via Ni^2＋-NTA column was higher than 98% with the specific activity 132.58 nmol C2H4/ （mg·h）.