噬夏孢欧文氏菌GGPP合成酶与八氢番茄红素合成酶相互作用的研究

Studies on the Interaction Between Geranylgeranyl Pyrophosphate Synthase and Phytoene Synthase of Erwinia Uredovora

GGPP合成酶与八氢番茄红素合成酶是噬夏孢欧文氏菌类胡萝卜素合成代谢中的两个关键酶。本研究将编码GGPP合成酶与八氢番茄红素合成酶的基因crtE和crtB共转化大肠杆菌或将crtE、crtB及编码八氢番茄红素脱氢酶的crtI共转化大肠杆菌,IPTG诱导其表达,然后利用亲和层析、凝胶过滤层析纯化重组蛋白。Western blotting分析结合N-末端序列分析表明,GGPP合成酶与八氢番茄红素合成酶以两种复合体的形式存在,而八氢番茄红素脱氢酶则不能与这两种酶形成稳定的复合体。

Geranylgeranyl pyrophosphate synthase（GGPP synthase） and phytoene synthase are two key enzymes in synthesis metabolism of carotenoids in Erwinia uredovora.In this study,crtE encoding GGPP synthase and crtB encoding phytoene synthase were co-transformed into E.coliBL21（DE3） to construct an engineering bacterium.Meanwhile,crtE,crtB as well as crtI encoding phytoene desaturase were also transformed into E.coli BL21（DE3） to construct another engineering bacterium.After induced by IPTG,expressed recombinant proteins were purified by affinity chromatography and gel filtration chromatography.GGPP synthase and phytoene synthase were found to have formed two complexes,while phytoene desaturase showed no interaction with these two enzymes.