Separation And Purification Of R-phycoerythrin Haitanensis Study Of The Physicochemical Properties And Biological Activity

R-PE,a major type of PE, is supramolecular protein-pigment complex,as light-harvesting pigment of red algae and cyanobacteria. PE,PC and APC are assembled into an organized rod-like cellular structure, the phycobilisome, attached to the stromal side of the thylakoid membrane,which is made up of light-harvesting system of marine alga.Optical energy is transferred successively from PE to PC then to APC,and finally to chlorophy II with an overall quantum efficiency of more than 95%,changed into chemcal energy at the photosystem.The crystal structure of several phycobiliproteins has been fefined to high resolution as well as R-PE, which can help explain the relation between struction and function of them.It's essential to study energy transfer and photosynthesis mechanism.R-PE is water-soluble and intensely orange-red fluorescent,used as fluorescent probe and a promising new photosentitizer,the recently hot area of photodynamic therapy because of the photodynamic characteristicsThis thesis optimized purification technics of R-PE from red algae, Porphyra haitanensis R-Phycoerythrin was isolated and purified from red alga, Porphyra haitanensis. After successive column chromatography on hydroxylapatite (HA) and DEAE-Sepharose Fast Flow ion-exchange column chromatography separations,the purity reach to reagent grade(A565nm/A280nm>5.3) This purification procedures are fast , simply and cost-effectively. Meanwhile, more effective measures were adopted to identify the purity of R-PE such as spectra index and agrose gel electrophoresis. Absorption spectra of purified R-PE were determined. The relative molecular weights of subunits consisting in R-PE were detected by SDS-PAGE .And then explore the stability of R-PE treated by various pH and temperature ,and its long-time storage.This job has real meaning for R-PE storage as goods transportation.Many phycobiliproteins have photodynamic bioactivity as photosentitizer to kill tumor cells. According to photodynamic difference between these three phycobiliproteins ( R-PE,R-PC and C-PC),The comparisons of the photodynamic effective of them on tumor cell Bcap-37 are showed firstly in the research. R-PC was also at first used as photosentitizer in photodynamic therapy(PDT). Since mammary cancer is one of women's popular disease,the Human mammary cancer line Bcap-37 is chosen specially to attempt to abtain some useful results for mammary cancer PDT. The studyings may be some theoretically and practicably valuable.