Bermejo Román R, Alvárez-Pez J M, Acién Fernández F G, Molina Grima E
Department of Physical and Analytical Chemistry, Jaen University, E.U.P. of Linares, Linares 23700, Spain.
J Biotechnol. 2002 Jan 31;93(1):73-85. doi: 10.1016/s0168-1656(01)00385-6.
Phycoerythrin is a major light-harvesting pigment of red algae and cyanobacteria that is widely used as a fluorescent probe and analytical reagent. In this paper, B-phycoerythrin and R-phycocyanin in native state, from the red alga Porphyridium cruentum were obtained by an inexpensive and simple process. The best results of this purification procedure were scaled up by a factor of 13 to a large preparative level using an anionic chromatographic column of DEAE cellulose. Gradient elution with acetic acid-sodium acetate buffer (pH 5.5) was used. In these conditions both 32% of B-phycoerythrin and 12% of R-phycocyanin contained in the biomass of the microalgae was recovered. B-phycoerythrin was homogeneous as determined by sodium dodecyl sulfate-poly-acrylamide gel electrophoresis (SDS-PAGE), yielding three migrating bands corresponding to its three subunits, consistent with the (alpha beta)(6)gamma subunit composition characteristic of this biliprotein and the spectroscopic characterization of B-PE (UV-visible absorption and emission spectroscopy; steady-state and polarization fluorescence), is accompanied. Finally, a preliminary cost analysis of the recovery process is presented.
藻红蛋白是红藻和蓝细菌的主要捕光色素,被广泛用作荧光探针和分析试剂。本文通过一种廉价且简单的方法,从红球藻中获得了天然状态的B-藻红蛋白和R-藻蓝蛋白。使用DEAE纤维素阴离子色谱柱,将该纯化过程的最佳结果放大了13倍,达到了大规模制备水平。采用乙酸-乙酸钠缓冲液(pH 5.5)进行梯度洗脱。在这些条件下,微藻生物质中含有的32%的B-藻红蛋白和12%的R-藻蓝蛋白被回收。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)测定,B-藻红蛋白是均一的,产生了对应于其三个亚基的三条迁移带,这与该双蛋白的(αβ)(6)γ亚基组成特征以及B-PE的光谱表征(紫外-可见吸收和发射光谱;稳态和偏振荧光)相符。最后,给出了回收过程的初步成本分析。