藻蓝蛋白α亚基裂合酶的分子克隆和酶动力学研究

摘要

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.%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.