蓝细菌光敏色素及藻红蛋白的生物合成分析-biosynthetic analysis of cyanobacteria photosensitive pigments and phycoerythrin.docxVIP

AbstractCyanobacteria are prokaryotes with the capability of oxygenic photosynthesis. Under the appropriate aquatic environment, a large amount of algae can grow which can induce water bloom. The process of photosynthesis is initiated by the absorption of light energy through phycobiliproteins. Phycobiliproteins are multi-subunit complexes that covalently bind a variety of linear tetrapyrrole pigments called bilins, enabling them to harvest light in the visible region of the spectrum. Lyases CpcE and CpcF specific for attaching PCB to Cys-84 of CpcA have been identified in several cyanobacteria. PecE and PecF catalyze both the attachment of PCB to PecA and its isomerization. More recently, the lyase CpcS has been identified as one of the lyases attaching PCB to the β-subunits of CPC and possibly allophycocyanin (ApcA, ApcB, ApcA2, ApcD, ApcF). The lyase CpeT1 can catalyze PCB s covalent attachment of the β subunit Cys-155 of CpcB and PecB.In this study, the entire pathway of the biosynthesis of holo-α and β-subunits of CPEfrom Gloeobacter violaceus PCC 7421 was reconstituted in Escherichia coli (E. coli). In PCC 7421, we found 20 genes which are homologous to the active lyase.The genes for the biosynthesis of apo-proteins (apo-CpeA or apo-CpeB), the genes ho1 and pebA and pebB encoding the enzymes for PEB production and the probable lyase genes for the attachment of PEB to apo-proteins were co-expressed in E. coli by a dual vector system. With the lyase CpeY, holo-αsubunits of CPE were acquired which showed spectroscopic properties similar to those of the same protein produced in cyanobacteria. The lyase CpeS2 can partially catalyze the attachment of PEB to CpeB.Zhao and his colleagues found that the lyase from Anabaena sp. PCC7120 cancatalyze PCB’s covalent attachment of α subunit Cys-82 and the β subunit Cys-80 of CPE from Calothrix sp PCC 7601. CPE in Gloeobacter violaceus PCC 7421 and Calothrix sp PCC 7601 showed high homology. We attempt to find the