Abstract
This study presents the development of a reliable method for quantifying C-phycocyanin. It was found that the spectrophotometric method commonly used for C-phycocyanin quantification tends to overestimate the actual amount of C-phycocyanin in Aphanizomenon flos-aquae (AFA) samples. The aim of this study was therefore to adapt the spectrophotometric C-phycocyanin quantification method specifically for A. flos-aquae, accounting for the variation in C-phycocyanin between different cyanobacteria species. High performance liquid chromatography (HPLC) was used as reference method and to avoid interference between molecules. The existing spectrophotometric equations for quantifying AFA C-phycocyanin were adapted using a C-phycocyanin standard. The method was then used to obtain a new set of spectrophotometric quantification equations adapted to the strain of interest and ensuring the accuracy of C-phycocyanin quantification while continuing to use a rapid, simple, and inexpensive method of pigment quantification. The method developed here could also be adapted to improve quantification methods for other types of phycocyanin, cyanobacteria, and other compounds of interest that are currently quantified by spectrophotometry.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
The datasets generated and analyzed in the current study are available from the corresponding author on reasonable request.
References
Benedetti S, Benvenuti F, Pagliarani S, Francogli S, Scoglio S, Canestrari F (2004) Antioxidant properties of a novel phycocyanin extract from the blue-green alga Aphanizomenon flos-aquae. Life Sci 75:2353–2362
Benedetti S, Rinalducci S, Benvenuti F, Francogli S, Pagliarani S, Giorgi L, Micheloni M, D’Amici GM, Zolla L, Canestrari F (2006) Purification and characterization of phycocyanin from the blue-green alga Aphanizomenon flos-aquae. J Chromatogr B 833:12–18
Bennett A, Bogorad L (1973) Complementary chromatic adaptation in a filamentous blue-green alga. J Cell Biol 58:419–435
Bhat VB, Madyastha KM (2000) C-phycocyanin: A potent peroxyl radical scavenger in vivo and in vitro. Biochem Biophys Res Commun 275:20–25
Everitt B (1998) The Cambridge dictionary of statistics. Cambridge University Press, Cambridge
Bryant DA, Guglielmi G, Tandeau de Marsac N, Castets A-M, Cohen-Baziere G (1979) The structure of cyanobacterial phycobilisomes: a model. Arch Microbiol 123:113–127
Busnel A (2018) Etude du potentiel de la cyanobactérie Aphanizomenon flos-aquae (AFA) pour des applications alimentaires. Doctoral Thesis, Nantes
Chen T, Wong Y-S, Zheng W (2006) Purification and characterization of selenium-containing phycocyanin from selenium-enriched Spirulina platensis. Phytochemistry 67:2424–2430
Cremonte M, Sisti D, Maraucci I, Giribone S, Colombo E, Rocchi MBL, Scoglio S (2017) The effect of experimental supplementation with the Klamath algae extract Klamin on attention-deficit/hyperactivity disorder. J Med Food 20:1233–1239
Glazer AN (1994) Phycobiliproteins — a family of valuable, widely used fluorophores. J Appl Phycol 6:105–112
Lauceri R, Bresciani M, Lami A, Morabito G (2018) Chlorophyll a interference in phycocyanin and allophycocyanin spectrophotometric quantification. J Limnol 77:1691
Menvielle-Bourg FJ, Vitacca A, Scoglio S (2011) L’algue Klamath et ses propriétés nutritionnelles. Un extrait spécifique aux effets neuromodulateurs et neuroprotecteurs. Phytothérapie 9:165–171
Merino JJ, Cabaña-Muñoz ME, Pelaz MJ (2020) The bluegreen algae (AFA) consumption over 48 hours increases the total number of peripheral CD34+ Cells in healthy patients: effect of short-term and long-term nutritional supplementation (curcumin/AFA) on CD34+ levels (blood). J Pers Med 10:49
Murakami A, Mimuro M, Ohki K, Fujita Y (1981) Absorption spectrum of allophycocyanin isolated from Anabaena cylindrica: variation of the absorption spectrum induced by changes of the physico-chemical environment. J Biochem (Tokyo) 89:79–86
Nichols HW, Bold HC (1965) Trichosarcina polymorpha gen. et sp. nov. J Phycol 1:34–38
Nowicka-Krawczyk P, Mühlsteinová R, Hauer T (2019) Detailed characterization of the Arthrospira type species separating commercially grown taxa into the new genus Limnospira (Cyanobacteria). Sci Rep 9:694
Padyana AK, Bhat VB, Madyastha KM, Rajashankar KR, Ramakumar S (2001) Crystal structure of a light-harvesting protein C-phycocyanin from Spirulina platensis. Biochem Biophys Res Commun 282:893–898
Rinalducci S, Roepstorff P, Zolla L (2009) De novo sequence analysis and intact mass measurements for characterization of phycocyanin subunit isoforms from the blue-green alga Aphanizomenon flos-aquae. J Mass Spectrom 44:503–515
Safari R, Amiri ZR, Kenari RE (2017) Antioxidant and antibacterial activities of C-phycocyanin from common name Spirulina platensis. Iran J Fish Sci 19:1911–1927
Seyidoglu N, Inan S, Aydin C (2017) A prominent superfood: Spirulina platensis. In: Shiomi N, Waisundara V (eds) Superfood and functionl food, Ch. 1. IntechOpen, Riejeka pp 1–27. https://doi.org/10.5772/66118
Soni B, Kalavadia B, Trivedi U, Madamwar D (2006) Extraction, purification and characterization of phycocyanin from Oscillatoria quadripunctulata—Isolated from the rocky shores of Bet-Dwarka, Gujarat, India. Process Biochem 41:2017–2023
Syrpas M, Bukauskaitė J, Ramanauskienė K, Karosienė JR, Majienė D, Bašinskienė L, Venskutonis PR (2020) Ultrasound-assisted extraction and assessment of biological activity of phycobiliprotein-rich aqueous extracts from wild cyanobacteria (Aphanizomenon flos-aquae). J Agric Food Chem 68:1896–1909
Thomas J-C (1989) L’antenne collectrice d’énergie lumineuse à phycobiliprotéines chez les cyanobactéries. Bull Soc Bot Fr Actual Bot 136:31–49
Zavřel T, Chmelík D, Sinetova MA, Červený J (2018) Spectrophotometric determination of phycobiliprotein content in cyanobacterium Synechocystis. J Vis Exp 139:58076
Zolla L, Bianchetti M (2001) High-performance liquid chromatography coupled on-line with electrospray ionization mass spectrometry for the simultaneous separation and identification of the Synechocystis PCC 6803 phycobilisome proteins. J Chromatogr A 912:269–279
Acknowledgements
We acknowledge the helpful discussions with Benoit Degrenne, Ph.D. and Jack Hoeniges, Ph.D. for correcting the English in this paper.
Funding
The authors thank the Pays de la Loire regional council for funding this work within the scientific framework of the applicable “Bone Health” chair.
Author information
Authors and Affiliations
Contributions
All authors contributed to the study conception and design. The materials were prepared by Delphine Drouin and Julie Billy, and data collected and analyzed by Julie Billy. The first draft of the manuscript was written by Julie Billy, and Olivier Goncalves commented on previous versions of the manuscript. All authors read and approved the final manuscript.
Corresponding author
Ethics declarations
Competing interests
The authors have no competing interests.
Additional information
Publisher's note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Billy, J., Pruvost, J., Lépine, O. et al. Development of a spectrophotometric method for quantification of C-phycocyanin in the cyanobacterium Aphanizomenon flos-aquae. J Appl Phycol 35, 1715–1726 (2023). https://doi.org/10.1007/s10811-023-03011-1
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10811-023-03011-1