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      Extracción y purificación de B-ficoeritrina de la microalga roja Rhodosorus marinus Translated title: Extraction and purification of B-phycoerythrin from the red microalga Rhodosorus marinus

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          Abstract

          En este trabajo se describe la purificación de la proteína B-ficoeritrina (B-PE) de la microalga roja Rhodosorus marinus. Inicialmente se extrajeron las ficobiliproteínas de las células microalgales mediante fragmentación manual y sonicación. La B-PE fue purificada mediante precipitación con sulfato de amonio y cromatografía de exclusión por tamaño e intercambio aniónico. Se determinó la pureza de B-PE por medio de índices y electroforesis en gel de poliacrilamida con dodecilsulfato de sodio. La caracterización espectroscópica de B-PE se realizó mediante espectroscopía UV-visible, espectroscopía de fluorescencia y dicroísmo circular. Rhodosorus marinus presentó tres tipos de ficobiliproteínas: ficoeritrina, ficocianina y aloficocianina. La B-PE purificada mostró un índice de pureza (A545/A280) de 4.8, picos característicos a 540 y 562 nm con un hombro a 498 nm, un máximo de emisión de fluorescencia de 578 nm, y una estructura secundaria casi estable ante cambios de pH. La B-PE mostró ser el pigmento predominante en R. marinus. Esta microalga roja podría ser una opción viable para la recuperación de esta cromoproteína.

          Translated abstract

          A description is given of the purification of B-phycoerythrin (B-PE) from the red microalga Rhodosorus marinus. Initially, phycobiliproteins were released from the microalgal cells by manual cellular fragmentation and sonication. B-PE was extracted with ammonium sulfate precipitation, and purified by anionic and size exclusion chromatography. Its purity was tested using indexes and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Spectroscopic characterization of B-PE was performed by UV-visible spectroscopy, fluorescence spectroscopy, and circular dichroism. Rhodosorus marinus showed three types of phycobiliproteins: phycoerythrin, phycocyanin, and allophycocyanin. The purified B-PE showed a purity ratio (A545/A280) of 4.8, characteristic peaks at 540 and 562 nm with a shoulder at 498 nm, fluorescence emission maximum at 578 nm, and a secondary structure almost stable with pH changes. B-PE was found to be the predominant pigment in R. marinus and this red microalga could be a viable option for the recovery of this chromoprotein.

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          Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4

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            A Rapid and Sensitive Method for the Quantitation of Microgram Quantities of Protein Utilizing the Principle of Protein-Dye Binding

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              Recovery of pure B-phycoerythrin from the microalga Porphyridium cruentum.

              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.
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                Author and article information

                Journal
                ciemar
                Ciencias marinas
                Cienc. mar
                Universidad Autónoma de Baja California, Instituto de Investigaciones Oceanológicas (Ensenada, Baja California, Mexico )
                0185-3880
                December 2009
                : 35
                : 4
                : 359-368
                Affiliations
                [03] Hermosillo Sonora orgnameUniversidad de Sonora orgdiv1Departamento de Investigaciones Científicas y Tecnológicas México mburboa@ 123456correom.uson.mx
                [02] Hermosillo Sonora orgnameUniversidad de Sonora orgdiv1Departamento de Investigación en Polímeros y Materiales orgdiv2Departamento de Física México
                [01] Hermosillo Sonora orgnameUniversidad de Sonora orgdiv1Departamento de Investigación en Polímeros y Materiales México
                Article
                S0185-38802009000400004 S0185-3880(09)03500400004
                ca7fca3b-48e9-4f28-b35c-ea14761503a9

                This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

                History
                : October 2009
                : May 2009
                Page count
                Figures: 0, Tables: 0, Equations: 0, References: 38, Pages: 10
                Product

                SciELO Mexico

                Categories
                Artículos

                microalgas,ficobiliproteínas,ficoeritrina,pigmentos,Rhodosorus marinus,microalgae,phycobiliproteins,phycoerythrin,pigments

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