黄花蒿法呢基焦磷酸合酶、紫穗槐二烯合酶双功能酶基因的构建、原核表达与功能鉴定

试验将青蒿素生物合成途径中催化两步连续反应的酶(法呢基焦磷酸合酶和紫穗槐二烯合酶)的基因进行融合,经大肠杆菌表达后鉴定其融合蛋白的功能。结果表明:融合蛋白具有了双功能酶活性。双功能酶基因的构建,为进一步将双功能酶基因转入黄花蒿,提高青蒿素含量奠定了基础。

Construction,E.coli expression and function identification of bifunctional enzymes fused by FPS and ADS from Artemisia annua L.

As the increasing demand of artemisinin has placed tremendous stress on artemisinin supplies worldwide in the treatment of malaria,more and more attention has been paid to increasing the content of artemisinin in Artemisia annua L..Fusion of farnesyl pyrophosphate synthase and amorpha-4,11-diene synthase genes,which catalyze two sequential reactions in biosynthesis pathway of artemisinin,was constructed and expressed in E.coli.Function analysis showed that the fused protein could catalyze the formation of amorpha-4,11-diene from GPP.The construction of bifunctional enzyme gene lays a foundation for further gene transformation into Artemisia annua L.to increase the content of artemisinin in the plant.