藻蓝蛋白α亚基裂合酶的分子克隆和酶动力学研究

Study on Molecular Cloning and Enzyme Kinetics of Anabaena α-Phycocyanin Lyase

为了比较研究不同藻种中藻蓝蛋白裂合酶CpcE/F的结构与功能的差异,对Anabaena sp.PCC 7120中的CpcE/F进行克隆,并进行大量表达,将表达的裂合酶CpcE/F用于藻蓝胆素(PCB)与Mastigocladus laminosus PCC 7603藻蓝蛋白α-亚基(α-PC)脱辅基蛋白(CpcA)的体外重组,得到天然活性的α-PC,从而表明CpcE/F所编码的蛋白质是α-PC生物合成的裂合酶,并对CpcE/F的酶动力学进行了初步研究。

To comparatively study the differences on structures and functions of phycocyanin lyase CpcE/F from different cyanobacteria,CpcE/F from Anabaena sp.PCC 7120 were cloned and expressed greatly.The reconstitution of PCB and CpcA from Mastigocladus laminosus PCC 7603 in vitro using the overexpressed lyase Anabaena sp.PCC 7120 CpcE/F showed that CpcE/F from Anabaena sp.PCC 7120 is the specific enzyme for biosynthesis of α-PC,furthermore,the enzyme kinetics of PcE/F lyase were studied preliminarily.