红藻多管藻(Polysiphonia Urceolata)R-藻蓝蛋白、别藻蓝蛋白吸收系数测定及藻胆体藻胆蛋白组成分析  

Light Absorption Coefficient Determination of R-phycocyanin and Allophycocyanin and Phycobiliptrotein Composition Analysis of Phycobilisomes from Polysiphonia Urceolata

作　　者：台丹丹 赵明日[1] 孙力[1] TAI Dan-dan;ZHAO Ming-ri;SUN Li(School of Life Sciences,Yantai University,Yantai 264005,China)

机构地区：[1]烟台大学生命科学学院,山东烟台264005

出　　处：《烟台大学学报（自然科学与工程版）》2021年第3期272-281,共10页Journal of Yantai University(Natural Science and Engineering Edition)

基　　金：山东省自然科学基金资助项目(ZR2018LD009)。

摘　　要：选用多维层析和非变性PAGE(Native-PAGE),从海生多管藻(Polysiphonia Urceolata)藻胆蛋白提取液中制备纯度符合光吸收系数测定要求的藻蓝(R-PC)和别藻蓝(AP)蛋白。选用Bradford和Hartree-Lowry法,用牛血清蛋白和γ-球蛋白作标准蛋白,对R-PC、AP进行蛋白浓度测定,并根据Lambert-Beer Law求得其在各自特征吸收峰处的光吸收系数(K)。选用更适合于多亚基复合体蛋白定量、以γ-球蛋白作标准的Hartree-Lowry法所获得的藻红蛋白(R-PE)、R-PC和AP的光吸收系数,建立了符合低浓度蛋白含量测定要求,可用特征吸收值(A)求得各藻胆蛋白含量比例的二元和三元线性方程组。藻胆蛋白组成分析表明:解离和完整藻胆体中R-PE、R-PC和AP物质的量比分别为7~9∶1∶1和11~13∶1∶3,这种组成差别应源于完整藻胆体中R-PC和AP的分子间相互作用,且有利于光能的高效单向传递。To measure the light absorption coefficients,R-phycocyanin(R-PC)and Allophycocyanin(AP)with purity requirement and the purity of the prepared R-PC and AP accord with the requirement are prepared from the solution of phycobiliproteins extracted from marine Polysiphonia urceolata by multidimension chromatography and native PAGE(Native-PAGE).The concentrations of the purified R-PC and AP are qualitatively determined by means of Bradford and Hartree-Lowry methods with bovine serum protein andγ-globulin as standard proteins,and the light absorption coefficients at their respective characteristic absorption peaks are calculated according to Lambert-Beer Law.The light absorption coefficients of R-phycoerythrin(R-PE),R-PC and AP obtained by the Lowry method withγ-globulin as standard protein are more suitable to measure qualitatively protein complexes with multi-subunit components.Two-variable and three-variable linear equations suitable for the determination of protein content and proportion in low concentration are established by taking the individual characteristic light absorbencies(Aλs)of R-PE,R-PC and AP as variables.The composition analysis demonstrates that in the disassociated phycobilisomes and in the intact phycobilisomes,the molar proportions of R-PE,R-PC and AP are 7-9∶1∶1 and 11-13∶1∶3,respectively.The differences of the molar proportions should be originated from the interaction between R-PC and AP in the intact phycobilisomes,which is favorable to the directional light energy transfer.

关 键 词：多管藻 藻胆蛋白 藻胆体 光吸收系数 蛋白定量 

分 类 号：Q946.1[生物学—植物学]