Abstract
Phycoerythrin is the major light-harvesting pigment-protein of the red algae Porphyridium cruentum and is widely used as fluorescent probe and analytical reagent. Additionally this protein has a potential application as natural dye in food industry. Nevertheless the knowledge of the functional properties of this alga protein is limited, hindering its application as food additive. In this article we report a biophysical characterization of B-phycoerythrin from Porphyridium cruentum (B-PE) in order to study its stability and spectral properties in a broad range of pHs. This information can help in its potential application as colorant in the food industry. Spectroscopic data obtained in this work show that B-PE has a stronger functional stability in the pH range 4.0–10.0, and Size Exclusion Chromatography suggests that the protein maintains a (αβ)6-γ oligomeric structure in that range of pHs. At pH 7.0, an apparent T m value of 77.5 ± 0.5 °C was calculated. At this pH, the protein is highly stable with a loss of only 20 % of its spectral properties (absorbance and fluorescence) after 25 days at room temperature. These results indicate that B-PE is more stable in a broad range of pHs than other phycoerythrin proteins, which would facilitate its use in the food industry.
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Notes
Since three different subunits are present in B-PE, the unfolding of this protein might be more complicated than a two-state mechanism.
References
D.B. Stengel, S. Connan, Z.A. Popper, Algal chemodiversity and bioactivity: sources of natural variability and implications for commercial application. Biotechnol. Adv. 29, 483–501 (2011)
A.C. Guedes, H.M. Amaro, F.X. Malcata, Microalgae as sources of high added-value compounds-a brief review of recent work. Biotechnol Progr 27, 597–613 (2011)
S. Sekar, M. Chandramohan, Phycobiliproteins as a commodity: trends in applied research, patents and commercialization. J. Appl. Phycol. 20, 113–136 (2008)
M.C. Santiago-Santos, T. Ponce-Noyola, R. Olvera-Ramírez, J. Ortega-López, R.O. Cañizares-Villanueva, Extraction and purification of phycocyanin from Calothrix sp. Process Biochem. 39, 2047–2052 (2004)
L.N. Liu, H.N. Su, S.G. Yan, S.M. Shao, B.B. Xie, X.L. Chen, X.Y. Zhang, B.C. Zhou, Y.Z. Zhang, Probing the pH sensitivity of R-phycoerythrin: investigations of active conformational and functional variation. Biochim. Biophys. Acta 1787, 939–946 (2009)
A.N. Glazer, Phycobiliproteins. Methods Enzymol. 167, 291–303 (1988)
E. Mörschel, E. Rhiel, in Phycobilisomes and Thylakoids, ed. by J.R. Harris, R.W. Horne (Academic Press, London, 1987), p. 210
G.J. Wedemayer, D.G. Kidd, A.N. Glazer, Cryptomonad biliproteins: bilin types and locations. Photosynth. Res. 48, 163–170 (1996)
A. Yaron and S. (M) Arad, Natural pigments from red microalgae for food and cosmetics. In: Charalambous G, editor. Food flavors, ingredients and composition. Amsterdam: Elsevier Science Publishers BV; 1993
M.F. de Jesus Raposo, R.M. de Morais, A.M. de Morais, Health applications of bioactive compounds from marine microalgae. Life Sci. 93, 479–486 (2013)
P. Spolaore, C. Joannis-Cassan, E. Duran, A. Isambert, Commercial applications of microalgae. J. Biosci. Bioeng. 101, 87–96 (2006)
Richa, V.K. Kannaujiya, M. Kesheri, G. Singh, R.P. Sinha, Biotechnological potentials of phycobiliproteins. Int. J. Pharmaceut. Res. Biosci. 2, B446–B454 (2011)
Q. Pan, M. Chen, J. Li, Y. Wu, C. Zhen, B. Liang, Antitumor function and mechanism of phycoerythrin from Porphyra haitanensis. Biol Res. 46, 87–95 (2013)
A.N. Glazer, C.S. Hixson, Subunit structure and chromophore composition of rhodophytan phycoerythrins. B-phycoerythrin and b-phycoerythrin. J. Biol. Chem. 252, 32–42 (1977)
R.W. Swanson, A.N. Glazer, Separation of phycobiliprotein subunits by reverse-phase high-pressure liquid chromatography. Anal. Biochem. 188, 295–299 (1990)
S.M. Wilbanks, A.N. Glazer, Rod structure of phycoerythrin II-containing phycobilisome II, Complete sequence and bilin attachment site of a phycoerythrin γ-subunit. J. Biol. Chem. 268, 1236–1241 (1993)
A. Camara-Artigas, J. Bacarizo, M. Andujar-Sanchez, E. Ortiz-Salmeron, C. Mesa-Valle, C. Cuadri, J.M. Martin-Garcia, S. Martinez-Rodriguez, T. Mazzuca-Sobczuk, M.J. Ibañez, J.P. Allen, pH-dependent structural conformations of B-phycoerythrin from Porphyridium cruentum. FEBS J. 279, 3680–3691 (2012)
D.J. Lundell, A.N. Glazer, R.J. de Lange, D.M. Brown, Bilin attachment sites in the α- and β-subunits of B-phycoerythrin. Amino acid sequence studies. J. Biol. Chem. 259, 5472–5480 (1984)
R. Bermejo Román, J.M. Alvárez-Pez, F.G. Acién Fernández, E. Molina Grima, Recovery of pure B-phycoerythrin from the microalga Porphyridium cruentum. J. Biotechnol. 93, 73–85 (2002)
R. Bermejo, E.M. Talavera, J.M. Álvarez Pez, Chromatographic purification and characterization of B-phycoerythrin from Porphyridium cruentum. Semipreparative high-performance liquid chromatographic separation and characterization of its subunits. J. Chrom. A. 917, 135–145 (2001)
R. Ficner, R. Huber, Refined crystal structure of phycoerythrin from Porphyridium cruentum at 0.23-nm resolution and localization of the gamma subunit. Eur. J. Biochem. 218, 103–106 (1993)
R. Bermejo, F.G. Acién, M.J. Ibáñez, J.M. Fernández, E. Molina, J.M. Alvarez-Pez, Preparative purification of B-phycoerythrin from the microalga Porphyridium cruentum by expanded-bed adsorption chromatography. J Chromatogr B Analyt Technol Biomed Life Sci. 790, 317–325 (2003)
R. Bermejo, D.J. Tobaruela, E.M. Talavera, A. Orte, J.M. Alvarez-Pez, Fluorescent behavior of B-phycoerythrin in microemulsions of aerosol OT/water/isooctane. J. Colloid Interface Sci. 263, 616–624 (2003)
S.K. Mishra, A. Shrivastav, I. Pancha, D. Jain, S. Mishra, Effect of preservatives for food grade C-Phycoerythrin, isolated from marine cyanobacteria Pseudanabaena sp. Int. J. Biol. Macromol. 47, 597–602 (2010)
M.J. Ibañez-Gonzalez, T. Mazzuca-Sobczuk, E. Molina-Grima, Procedimiento de purificación de biomoléculas que utiliza cromatografía de adsorción en lecho expandido. Patent ES 2383866, B1 (2013)
M.I. Muro-Pastor, F.N. Barrera, J.C. Reyes, F.J. Florencio, J.L. Neira, The inactivating factor of glutamine synthetase, IF7, is a “natively unfolded” protein. Protein Sci. 12, 1443–1454 (2003)
A. Gaigalas, T. Gallagher, K. Cole, T. Singh, L. Wang, Y. Zhang, A multistate model for the fluorescence response of R-phycoerythrin. Photochem. Photobiol. 82, 635–644 (2006)
M.A. Andrade, P. Chacón, J.J. Merelo, F. Morán, Evaluation of secondary structure of proteins from UV circular dichroism using an unsupervised learning neural network. Prot. Eng. 6, 383–390 (1993)
A. Orta-Ramirez, J.E. Merrill, D.M. Smith, pH affects the thermal inactivation parameters of R-phycoerythrin from Porphyra yezoensis. J. Food Sci. 65, 1046–1050 (2000)
L.G. Roth, D.S. Berns, C.H. Chen, Comparative thermodynamic elucidation of the structural stability of thermophilic proteins. Biophys. Chem. 60, 89–97 (1996)
C. Abad-Zapatero, J.L. Fox, M.L. Hackert, The quaternary structure of a unique phycobiliprotein: B-phycoerythrin from Porphyridium cruentum. Biochem. Biophys. Res. Commun. 78, 266–272 (1977)
A.N. Glazer, C.S. Hixson, Characterization of R-phycocyanin. Chromophore content of R-phycocyanin and C-phycoerythrin. J. Biol. Chem. 250, 5487–5495 (1975)
C.N. Pace, G.R. Grimsley, J.M. Scholtz, Protein ionizable groups: pK values and their contribution to protein stability and solubility. J. Biol. Chem. 284, 13285–13289 (2009)
Acknowledgments
This work was supported by the Ministerio de Ciencia e Innovación (MCINN) BIO2009-13261-C02-01/02 and Ministerio de Economia y competitividad (MECOM) BIO2012-39922-C02-01/02 of Spain and FEDER (EU). Additional funding was supported by grant P09-CVI-5063 from Andalusian Regional Government (Spain) and FEDER (EU). This work has been performed by members of the research groups BIO-328 Protein Structures of the Andalusian Regional Government (Spain).
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González-Ramírez, E., Andújar-Sánchez, M., Ortiz-Salmerón, E. et al. Thermal and pH Stability of the B-Phycoerythrin from the Red Algae Porphyridium cruentum . Food Biophysics 9, 184–192 (2014). https://doi.org/10.1007/s11483-014-9331-x
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DOI: https://doi.org/10.1007/s11483-014-9331-x