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Purification and characterization of phycoerythrin from the Caspian Sea red macroalga (Osmundea caspica)

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Abstract

Phycoerythrin (R-PE) is a naturally occurring colorant with excellent fluorescent and antioxidant properties with a wide range of applications. This study aimed to compare the use of different purification methods, which are activated charcoal (AC), ammonium sulfate precipitation (ASP), or anion-exchange chromatography (IE) alone and a combination of AC, ASP, and IE, for R-PE extraction from the Caspian Sea red macroalga (Osmundea caspica). Response surface methodology (RSM) was employed for preliminary purification, with the independent variables being activated charcoal content (0.1–1% w/v) and stirring time (2–10 min). With an AC content of 0.4% and a stirring time of 2 min, R-PE purity and concentration were optimized. The predicted values from the equations agreed well with the experimental values, demonstrating the model's robustness. A three-step increase in the ionic strength of IE was also evaluated at three different treatments (PECE, PECE + AC + IE, and ASP + IE). The highest R-PE purity and lowest Fluorescence intensity and antioxidant activity were obtained in the fraction of AE-200 (second fraction) with an index of 2.8 and a recovery yield of 48% in ASP + IE, whereas an index purity of 2 and the highest recovery yield of 67% were obtained using PECE + AC + IE.

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All data generated or analyzed during this study are included in this published article and its supplementary information file.

References

  • Baghel RS, Trivedi N, Gupta V, Neori A, Reddy CRK, Lali A, Jha B (2015) Biorefining of marine macroalgal biomass for production of biofuel and commodity chemicals. Green Chem 17:2436-2443

  • Baghel RS, Reddy C, Jha B (2014) Characterization of agarophytic seaweeds from the biorefinery context. Bioresour Technol 159:280–285

    Article  CAS  Google Scholar 

  • Blas-Valdivia V, Rojas-Franco P, Serrano-Contreras JI, Sfriso AA, Garcia-Hernandez C, Franco-Colín M, Cano-Europa E (2021) C-phycoerythrin from Phormidium persicinum prevents acute kidney injury by attenuating oxidative and endoplasmic reticulum stress. Mar Drugs 19:589

    Article  CAS  Google Scholar 

  • Bermejo R, Ruiz E, Ramos A, Acién FG (2013) Pilot-scale recovery of phycoerythrin from Porphyridium cruentum using expanded bed adsorption chromatography. Sep Sci Technol 48:1913–1922

    Article  CAS  Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  CAS  Google Scholar 

  • Burits M, Bucar F (2000) Antioxidant activity of Nigella sativa essential oil. Phytother Res 14:323–328

    Article  CAS  Google Scholar 

  • Chu WL (2012) Biotechnological applications of microalgae. IeJSME 6:24-37

  • Chuner C, Chunxia L, Shuxian W, Qing W, Ziye G, Peimin H (2012) Large scale preparation of phycobiliproteins from Porphyra yezoensis using co-precipitation with ammonium sulfate. Nat Sci 4:536–543

    Google Scholar 

  • Dastkhoon M, Ghaedi M, Asfaram A, Goudarzi A, Langroodi SM, Tyagi I (2015) Ultrasound assisted adsorption of malachite green dye onto ZnS: Cu-NP-AC: equilibrium isotherms and kinetic studies–response surface optimization. Sep Purif Technol 156:780–788

    Article  CAS  Google Scholar 

  • de Amarante MCA, Braga ARC, Sala L, Kalil SJ (2020) Colour stability and antioxidant activity of C-phycocyanin-added ice creams after in vitro digestion. Food Res Int 137:963–969

    Google Scholar 

  • Uju, Dewi NPSUK, Santoso J, Setyaningsih I, Hardingtyas S, Yopi (2020). Extraction of phycoerythrin from Kappaphycus alvarezii seaweed using ultrasonication. IOP Conf Ser: Earth Environ Sci 414:012028

  • Hilditch C, Balding P, Jenkins R, Smith A, Rogers L (1991) R-phycoerythrin from the macroalga Corallina officinalis (Rhodophyceae) and application of a derived phycofluor probe for detecting sugar-binding sites on cell membranes. J Appl Phycol 3:345–54

  • Denis C, Massé A, Fleurence J, Jaouen P (2009) Concentration and pre-purification with ultrafiltration of a R-phycoerythrin solution extracted from macro-algae Grateloupia turuturu: Process definition and up-scaling. Sep Purif Technol 69:37–42

    Article  CAS  Google Scholar 

  • Fekrat F, Nami B, Ghanavati H, Ghaffari A, Shahbazi M (2019) Optimization of chitosan/activated charcoal-based purification of Arthrospira platensis phycocyanin using response surface methodology. J Appl Phycol 31:1095–1105

    Article  CAS  Google Scholar 

  • Fitzgerald C, Gallagher E, Tasdemir D, Hayes M (2011) Heart health peptides from macroalgae and their potential use in functional foods. J Agric Food Chem 59:6829–6836

    Article  CAS  Google Scholar 

  • Galland-Irmouli A, Pons L, Lucon M, Villaume C, Mrabet N, Guéant J (2000) One-step purification of R-phycoerythrin from the red macroalga Palmaria palmata using preparative polyacrylamide gel electrophoresis. J Chromatogr B 739:117–123

    Article  CAS  Google Scholar 

  • Gao S, Wang W, Wang Y, Luo G, Dai Y (2010) Influence of alcohol treatments on the activity of lipases immobilized on methyl-modified silica aerogels. Bioresour Technol 101:7231–7238

    Article  CAS  Google Scholar 

  • García AB, Longo E, Murillo MC, Bermejo R (2021) Using a B-Phycoerythrin extract as a natural colorant: application in milk-based products. Molecules 26:297

  • Ghosh T, Mishra S (2020) Studies on extraction and stability of C-phycoerythrin from a marine cyanobacterium. Front Sust Food Syst 4:102

    Article  Google Scholar 

  • Gupta A, Sainis JK (2010) Isolation of C-phycocyanin from Synechococcus sp., (Anacystis nidulans BD1). J Appl Phycol 22:231–233

    Article  CAS  Google Scholar 

  • Isailovic D (2005) Subunits of highly fluorescent protein R-phycoerythrin as probes for cell imaging and single-molecule detection. PhD Thesis, Iowa State University

  • Kaixian Q, Franklin M, Borowitzka MA (1993) The study for isolation and purification of R-phycoerythrin from a red alga. Appl Biochem Biotech 43:133–139

    Article  Google Scholar 

  • Kanatt SR, Rao M, Chawla S, Sharma A (2013) Effects of chitosan coating on shelf-life of ready-to-cook meat products during chilled storage. LWT-Food Sci Technol 53:321–326

    Article  CAS  Google Scholar 

  • Karpinsky MG, Shiganova TA, Katunin DN (2005) Introduced species. In: Kostianoy AG, Kosarev AN (eds) The Caspian Sea Environment. Springer, Berlin, pp 175–190

    Chapter  Google Scholar 

  • Lauceri R, Zittelli GC, Maserti B, Torzillo G (2018) Purification of phycocyanin from Arthrospira platensis by hydrophobic interaction membrane chromatography. Algal Res 35:333–340

    Article  Google Scholar 

  • Lauceri R, Zittelli GC, Torzillo G (2019) A simple method for rapid purification of phycobiliproteins from Arthrospira platensis and Porphyridium cruentum biomass. Algal Res 44:101685

    Article  Google Scholar 

  • Lee S-H, Lee JE, Kim Y, Lee S-Y (2016) The production of high purity phycocyanin by Spirulina platensis using light-emitting diodes based two-stage cultivation. Appl Biochem Biotech 17:3823–3895

    Google Scholar 

  • Li W, Su H-N, Pu Y, Chen J, Liu L-N, Liu Q, Qin S (2019) Phycobiliproteins: Molecular structure, production, applications, and prospects. Biotechnol Adv 37:340–353

    Article  CAS  Google Scholar 

  • Liao X, Zhang B, Wang X, Yan H, Zhang X (2011) Purification of C-phycocyanin from Spirulina platensis by single-step ion-exchange chromatography. Chromatographia 73:291–296

    Article  CAS  Google Scholar 

  • Liu J-Y, Jiang T, Zhang J-P, Liang D-C (1999) Crystal structure of allophycocyanin from red algae Porphyra yezoensis at 2.2-Å resolution. J Biol Chem 274:16945–16952

    Article  CAS  Google Scholar 

  • Liu L-N, Chen X-L, Zhang X-Y, Zhang Y-Z, Zhou B-C (2005) One-step chromatography method for efficient separation and purification of R-phycoerythrin from Polysiphonia urceolata. J Biotech 116:91–100

    Article  CAS  Google Scholar 

  • Liu Y, Feng Y, Lun J (2012) Aqueous two-phase countercurrent distribution for the separation of c-phycocyanin and allophycocyanin from Spirulina platensis. Food Bioprod Process 90:111–117

    Article  CAS  Google Scholar 

  • Malairaj S, Muthu S, Gopal VB, Perumal P, Ramasamy R (2016) Qualitative and quantitative determination of R-phycoerythrin from Halymenia floresia (Clemente) C. Agardh by polyacrylamide gel using electrophoretic elution technique. J Chromatogr A 1454:120–126

    Article  CAS  Google Scholar 

  • Manirafasha E, Murwanashyaka T, Ndikubwimana T, Yue Q, Zeng X, Lu Y (2017) Ammonium chloride: a novel effective and inexpensive salt solution for phycocyanin extraction from Arthrospira platensis. J Appl Phycol 29:1261–1270

    Article  CAS  Google Scholar 

  • Mehdipour N, Gerami MH, Nemati H, Rahnama R, Hamzehpour A (2015) Spatial and temporal variation of macroalgae along the southern coasts of the Caspian Sea relating to environmental parameters. J Persian Gulf 6:65–73

    Google Scholar 

  • Mehdipour N, Sheijooni Fumani N, Rahnama R (2014) Proximate and fatty acid composition of the southern Caspian Sea macroalgae. J Persian Gulf 5:63–72

    Google Scholar 

  • Minkova K, Gigova L, Tchernov A, Stojanova M, Ivanova N, Boteva R (2007) Isolation of pure C-phycocyanin from Arthrospira maxima and Arthrospira fusiformis by a modified non-chromatographic rivanol sulfate procedure. C R Acad Bulgare Sci 60:795–798

    CAS  Google Scholar 

  • Mittal R, Tavanandi HA, Mantri VA, Raghavarao KSMS (2017) Ultrasound assisted methods for enhanced extraction of phycobiliproteins from marine macro-algae, Gelidium pusillum (Rhodophyta). Ultrason Sonochem 38:92–103

    Article  CAS  Google Scholar 

  • Mittal R, Sharma R, Raghavarao K (2019) Aqueous two-phase extraction of R-phycoerythrin from marine macro-algae, Gelidium pusillum. Bioresour Technol 280:277–286

    Article  CAS  Google Scholar 

  • Moshfegh A, Salehzadeh A, Sadat Shandiz SA, Shafaghi M, Naeemi AS, Salehi S (2019) Phytochemical analysis, antioxidant, anticancer and antibacterial properties of the Caspian Sea red macroalgae, Laurencia caspica. Iran J Sci Technol Transact A 43:49–56

    Article  Google Scholar 

  • Moraes CC, Kalil SJ (2009) Strategy for a protein purification design using C-phycocyanin extract. Bioresour Technol 100:5312–5317

    Article  CAS  Google Scholar 

  • Munier M, Jubeau S, Wijaya A, Morancais M, Dumay J, Marchal L, Fleurence J (2014) Physicochemical factors affecting the stability of two pigments: R-phycoerythrin of Grateloupia turuturu and B-phycoerythrin of Porphyridium cruentum. Food Chem 150:400–407

    Article  CAS  Google Scholar 

  • Munier M, Morancais M, Dumay J, Jaouen P, Fleurence J (2015) One-step purification of R-phycoerythrin from the red edible seaweed Grateloupia turuturu. J Chromatogr B 992:23–29

    Article  CAS  Google Scholar 

  • Niu JF, Wang GC, Zhou BC, Lin XZ, Chen CS (2007) Purification of R-phycoerythrin from Porphyra haitanensis (Bangiales, Rhodophyta) using expanded-bed absorption. J Phycol 43:1339–1347

    Article  CAS  Google Scholar 

  • Nguyen HPT, Morançais M, Déléris P, Fleurence J, Nguyen-Le CT, Vo KH (2020) Purification of R-phycoerythrin from a marine macroalga Gracilaria gracilis by anion-exchange chromatography. J Appl Phycol 32:553–561

    Article  Google Scholar 

  • Pan-utai W, Iamtham S (2019) Extraction, purification and antioxidant activity of phycobiliprotein from Arthrospira platensis. Process Biochem 82:189–198

    Article  CAS  Google Scholar 

  • Patil G, Chethana S, Sridevi A, Raghavarao K (2006) Method to obtain C-phycocyanin of high purity. J Chromatogr A 1127:76–81

    Article  CAS  Google Scholar 

  • Pereira T, Barroso S, Mendes S, Amaral RA, Dias JR, Baptista T (2020) Optimization of phycobiliprotein pigments extraction from red algae Gracilaria gracilis for substitution of synthetic food colorants. Food Chem 321:1–9

    Article  Google Scholar 

  • Ramos A, Acien FG, Fernandez-Sevilla JM, Gonzalez CV, Bermejo R (2011) Development of a process for large-scale purification of C-phycocyanin from Synechocystis aquatilis using expanded bed adsorption chromatography. J Chromatogr B 879:511–519

    Article  CAS  Google Scholar 

  • Rastogi RP, Sonani RR, Madamwar D (2015) Physico-chemical factors affecting the in vitro stability of phycobiliproteins from Phormidium rubidum A09DM. Bioresour Technol 190:219–226

    Article  CAS  Google Scholar 

  • Rossano R, Ungaro N, D’Ambrosio A, Liuzzi G, Riccio P (2003) Extracting and purifying R-phycoerythrin from Mediterranean red algae Corallina elongata Ellis & Solander. J Biotechnol 101:289–293

    Article  CAS  Google Scholar 

  • Rousseau F, Gey D, Kurihara A, Maggs CA, Martin-Lescanne J, Payri C (2017) Molecular phylogenies support taxonomic revision of three species of Laurencia (Rhodomelaceae, Rhodophyta), with the description of a new genus. Eur J Taxon 269:1–19

    Google Scholar 

  • Santiago-Santos MC, Ponce-Noyola T, Olvera-Ramı́rez R, Ortega-López J, Cañizares-Villanueva RO (2004) Extraction and purification of phycocyanin from Calothrix sp. Process Biochem 39:2047-2052

  • Sampath-Wiley P, Neefus CD (2007) an improved method for estimating R-phycoerythrin and R-phycocyanin contents from crude aqueous extracts of Porphyra (Bangiales, Rhodophyta). J Appl Phycol 19:123–129

    Article  CAS  Google Scholar 

  • Schneider G, Figueroa FL, Vega J, Avilés A, Chaves P, Horta PA, Bonomi-Barufi J (2020) Physiological and biochemical responses driven by different UV-visible radiation in Osmundea pinnatifida (Hudson) Stackhouse (Rhodophyta). Photochem Photobiol Sci 19:1650–1664

    Article  CAS  Google Scholar 

  • Sekar S, Chandramohan M (2008) Phycobiliproteins as a commodity: trends in applied research, patents and commercialization. J Appl Phycol 20:113–136

    Article  Google Scholar 

  • Senthilkumar N, Kurinjimalar C, Thangam R, Suresh V, Kavitha G, Gunasekaran P (2013a) Further studies and biological activities of macromolecular protein R-phycoerythrin from Portieria hornemannii. Int J Biol Macromolec 62:107–116

  • Senthilkumar N, Suresh V, Thangam R, Kurinjimalar C, Kavitha G, Murugan P (2013b) Isolation and characterization of macromolecular protein R-phycoerythrin from Portieria hornemannii. Int J Biol Macromol 55:150–160

  • Soni B, Trivedi U, Madamwar D (2008) A novel method of single step hydrophobic interaction chromatography for the purification of phycocyanin from Phormidium fragile and its characterization for antioxidant property. Bioresour Technol 99:188–194

    Article  CAS  Google Scholar 

  • Sudhakar M, Saraswathi M, Nair BB (2014) Extraction, purification and application study of R-phycoerythrin from Gracilaria corticata (J. Agardh) J. Agardh var. corticata. Indian J Nat Prod Resour 5:371–374

    Google Scholar 

  • Sun L, Wang S, Gong X, Zhao M, Fu X, Wang L (2009) Isolation, purification and characteristics of R-phycoerythrin from a marine macroalga Heterosiphonia japonica. Protein Express Purif 64:146–154

    Article  CAS  Google Scholar 

  • Tahmasebi M, Mofid V (2021) Innovative synbiotic fat-free yogurts enriched with bioactive extracts of the red macroalgae Laurencia caspica: formulation optimization, probiotic viability, and critical quality characteristics. J Food Meas Charact 15:4876–4887

    Article  Google Scholar 

  • Vandanjon L, Jaouen P, Rossignol N, Quéméneur F, Robert JM (1999) Concentration and desalting by membrane processes of a natural pigment produced by the marine diatom Haslea ostrearia Simonsen. Progr Indust Microbiol 35:393–402

    Article  CAS  Google Scholar 

  • Ventura SPM, de Barros RLF, de Pinho Barbosa JM, Soares CMF, Lima AS, Coutinho JAP (2012) Production and purification of an extracellular lipolytic enzyme using ionic liquid-based aqueous two-phase systems. Green Chem 14:734–740

    Article  CAS  Google Scholar 

  • Wang L, Wang S, Fu X, Sun L (2015) Characteristics of an R-phycoerythrin with two γ subunits prepared from red macroalga Polysiphonia urceolata. PLoS ONE 10:1–15

    Google Scholar 

  • Yabuta Y, Fujimura H, Kwak CS, Enomoto T, Watanabe F (2010) Antioxidant activity of the phycoerythrobilin compound formed from a dried Korean purple laver (Porphyra sp.) during in vitro digestion. Food Sci Technol Res 16:347–352

    Article  CAS  Google Scholar 

  • Zhou H, Chen X, Wang C, Ye J, Chen H (2012) Purification and characterization of a novel ~18 kDa antioxidant protein from Ginkgo biloba seeds. Molecules 17:14778–14794

    Article  CAS  Google Scholar 

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Acknowledgements

The author wishes to thank the center of Keyvan Marine Science and Technology Research Center (Guilan province, Iran) managers for their support and provision of the facilities and Ali Sadeghpour for their kind help. Leonel Pereira thanks to MARE, Marine and Environmental Sciences Centre, and to the Associate Laboratory ARNET. Moreover, a special thank goes to the Iranian Plant Antioxidants Center of Excellence for scientific advice in this research.

Funding

This research activities were carried out under the support of Iran National Science Foundation, INSF, Iranian Deputy of Science and Technology.

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Authors and Affiliations

  1. Keyvan Marine Science and Technology Research Center, Islamic Azad University (IAU), Lahijan Branch, Lahijan, Iran

    Hamid Eshaghzadeh

  2. Department of Environmental Sciences, Gorgan University of Agricultural Sciences and Natural Resources, P.O. Box 4913815739, Gorgan, Iran

    Maryam Shahbazi

  3. Nelson Marlborough Institute of Technology, H-Block, 322 Hardy Street, Private Bag 19, Nelson, 7042, New Zealand

    Sobhan Rana Akhavan

  4. MARE-Marine and Environmental Sciences Centre, Department of Life Sciences, University of Coimbra, 3001-456, Coimbra, Portugal

    Leonel Pereira

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  1. Hamid Eshaghzadeh
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Contributions

All of the experiments, data analysis, and manuscript preparation were conducted by Hamid Eshaghzadeh. In addition, Sobhan Akhavan assisted in macroalgae collection and laboratory activities, Maryam Shahbazi and Leonel Pereira were responsible for the conceptualization and supervision of the experiments and the critical revision of the manuscript.

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Eshaghzadeh, H., Shahbazi, M., Akhavan, S.R. et al. Purification and characterization of phycoerythrin from the Caspian Sea red macroalga (Osmundea caspica). J Appl Phycol 35, 317–330 (2023). https://doi.org/10.1007/s10811-022-02882-0

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