Bioprospecting of Spirulina biomass using novel extraction method for the production of C-Phycocyanin as effective food colourant


Research Articles | Published:

Print ISSN : 0970-4078.
Online ISSN : 2229-4473.
Pub Email:
Doi: 10.1007/s42535-021-00320-z
First Page: 484
Last Page: 492
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Keywords: n Spirulina sp., Phycobiliprotein, C-Phycocyanin, Natural colourant, Alum


Spirulina sp. MCCABT-SP0001 was grown in a miniature raceway pond system: an innovative and low-cost cultivation method for extraction of the natural pigment C-Phycocyanin. The extractions of C-Phycocyanin from Spirulina sp. were carried out by three different methods and the extracted C-phycocyanin was estimated separately for each method. Among the different methods of extraction, Freezing and Thawing, Alum and 15 M HCl extraction showed the best results, i.e., 0.40 mg/ml, 0.21 mg/ml and 0.19 mg/ml respectively. The C-Phycocyanin was partially purified using Ammonium Sulphate precipitation and later dialysed. The molecular weight of the powdered C-Phycocyanin was determined using SDS-PAGE. The molecular weight of C-Phycocyanin was found to be α14 kDa and β15 kDa subunits. The partially purified C-Phycocyanin was investigated for HPLC to know the presence of vitamins, both qualitatively and quantitatively. The vitamins present in C-Phycocyanin were Niacin (8.7 mg/ml), Pyridoxine (0.38 mg/ml), Thiamine (0.434 mg/ml) and Riboflavin (0.11 mg/ml). The partially purified C-Phycocyanin was examined for its staining ability cum stability with boiled rice as a natural colourant. Our research analysis findings suggest that C-Phycocyanin can be used as an alternative to synthetic food colourants, with high nutritional values for better health.

n                     Spirulina sp., Phycobiliprotein, C-Phycocyanin, Natural colourant, Alum

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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. 137:109602

Aslam J, Mohajir MS, Khan SA, Khan AQ (2008) HPLC analysis of water-soluble vitamins (B1, B2, B3, B5, B6) in in vitro and ex vitro germinated chickpea (Cicer arietinum L. ). Afr J Biotehnol 7(14):2310–2314

Belay A, Yoshimichi O, Miyakawa K, Shimamatsu H (1993) Current knowledge on potential health benefits of Spirulina. J Appl Phycol 5(2):235–241

Bennett A, Bogorad L (1973) Comparative chromatic adaptation in a filamentous blue-green alga. J Cell Biol 58:419–435

Bermejo-Bescos P, Pinero-Estrada E, Villar Del Fresno AM (2008) Neuroprotection by Spirulina platensis protean extract and phycocyanin against iron-induced toxicity in SH-SY5Y neuroblastoma cells. Toxicol in Vitro 22:1496–1502

Choi WY, Lee HY (2018) Effect of ultrasonic extraction on production and structural changes of C-phycocyanin from marine Spirulina maxima. Int J Mol Sci 19(1):220

Deng R, Chow TJ (2010) Hypolipidemic, antioxidant, and antiinflammatory activities of microalgae Spirulina. Cardiovasc Ther 28:33–45

Elumalai S, Rajesh KG, Jegan G, Sangeetha T, Roop SD (2014) Extraction of phycocyanin an important pharmaceutical phycobiliproteins from cyanobacteria. Int J Pharm Res Dev 6(4):67–74

Grahl S, Strack M, Weinrich R, Mörlein D (2018) Consumer-oriented product development: the conceptualization of novel food products based on Spirulina (Arthrospira platensis) and resulting consumer expectations. J Food Qual.

Grahl S, Strack M, Mensching A, Morlein D (2020) Alternative protein sources in Western diets: food product development and consumer acceptance of Spirulina filled pasta. Food Qual. Prefer. 84:103933.

Gupta A, Sainis JK (2010) Isolation of C-phycocyanin from Synechococcus sp., (Anacystis nidulans BD1). J Appl Phycol 22(3):231–233

Hadiyanto H, Harjanto GD, Huzain M, Aji RW (2019a) Production of antioxidant C-phycocyanin using extraction process of Spirulina platensis in large scale industry. In IOP Conf Ser Mater Sci Eng 633 (1): 012025. IOP Publishing

Hadiyanto H, Aziz ML, Joelyna FA, Sa’Adah AN, Putra MS, Filardli AM, Suzery M (2019b) The nutritional enrichment of dried noodles by using phycocyanin extracted from Spirulina sp as an effort of food fortification. In J Phys Conf Series 1295 (1): 012036. IOP Publishing

Hussein A, Ibrahim G, Kamil M, El-Shamarka M, Mostafa S, Mohamed D (2021) Spirulina-enriched pasta as functional food rich in protein and antioxidant. Biointerface Res Appl Chem 11:14736–14750

Karthikeyan D, Muthukumaran M, Balakumar BS (2016) Mass cultivation of microalgae in open raceway pond for biomass and biochemicals. Produc Int J Adv Res Biol Sci 3(2):247–260

Kuddus M, Singh P, Thomas G, Al-Hazimi A (2013) Recent developments in production and biotechnological applications of C-phycocyanin. Biomed Res Int 3:742859

Kumar RR, Rao PH, Subramanian VV, Sivasubramanian V (2014a) a) Enzymatic and non-enzymatic antioxidant potentials of Chlorella vulgaris grown in effluent of a confectionery industry. J Food Sci Technol 51(2):322–328

Kumar D, Dhar DW, Pabbi S, Kumar N, Walia S (2014b) b) Extraction and purification of C-phycocyanin from Spirulina platensis (CCC540). Ind J Plant Physiol 19(2):184–188

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(3–4):291–296

Mahfooz S, Bano S, Shamim A, Husain A, Farooqui A (2017) Partial purification, characterization and bioactive potential of C-phycocyanin from cyanobacterium Plectonema boryanum. Biochem Cell Arch 17(1):57–64

Manirafasha E, Murwanashyaka T, Ndikubwimana T, Yue Q, Zeng X, Lu Y, Jing K (2017) Ammonium chloride: a novel effective and inexpensive salt solution for phycocyanin extraction from Arthrospira (Spirulina platensis). J Appl Phycol 29(3):1261–1270

MonaliGhurde U, Padwad MM, Deshmukh VR, Malode SN (2016) Extraction of natural dye from Ixora coccinea (Linn.) flowers for cotton fabric colouration. Int J Sci Res 5(1):1272–1276

Moraes C, Sala L, Cerveira G, Kalil S (2011) C-phycocyanin extraction from Spirulina platensis wet biomass. Braz J Chem Eng 28:45–49

Nouri E, Abbasi H, Rahimi E (2018) Effects of processing on stability of water- and fat-soluble vitamins, pigments (C-phycocyanin, carotenoids, chlorophylls) and colour characteristics of Spirulina platensis. Qual Assur Saf Crop 10:1–16

Pan-utai W, Kahapana W, Iamtham S (2018) Extraction of C-phycocyanin from Arthrospira (Spirulina) and its thermal stability with citric acid. J Appl Phycol 30(1):231–242

Prabuthas P, Majumdar S, Srivastav PP, Mishra HN (2011) Standardization of rapid and economical method for neutraceuticals extraction from algae. J Stored Prod Postharvest Res 2(5):93–96

Pradeep HN, Nayak CA (2019) Enhanced stability of C-phycocyanin colorant by extrusion encapsulation. J Food Sci Technol 56(10):4526–4534

Ramakrishnan R (2013) anticancer properties of blue green algae SpirulinaPlatensis -a review. Int J Curr Med Pharm 3(4):159–168

Romay C, Gonzalez R (2000) Phycocyanin is an antioxidant protector of human erythrocytes against lysis by peroxyl radicals. J Pharm Pharmacol 52(4):367–368

Sambrook J, Russell DW (2001) SDS polyacrylamide gel electrophoresis for proteins. Commonly used techniques in molecular cloning, in 3rd edition (eds. Sambrook and Russell) Molecular Cloning, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, pp A840-A851

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

Sarada R, Pillai MG, Ravishankar G (1999) Phycocyanin from Spirulina sp: influence of processing of biomass on Phycocyanin yield, analysis of efficacy of extraction methods and stability studies on phycocyanin. Process Biochem 34:795–801

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

Sharoba AM (2014) Nutritional value of spirulina and its use in the preparation of some complementary baby food formulas. J Agroaliment Proc Technol 20(4):330–350

Silveira ST, Burkert JDM, Costa JAV, Burkert CAV, Kalil SJ (2007) Optimization of phycocyanin extraction from Spirulina platensis using factorial design. Bioresour Technol 98(8):1629–1634

Singh P, Kuddus M, Thomas G (2010) An efficient method for extraction of C-phycocyanin from Spirulina sp. and its binding affinity to blood cells, nuclei and genomic DNA. Int Res J Biotechnol 1(5):080–085

Sivasankari S, Naganandhini N, Ravindran D (2014) Comparison of different extraction methods for Phycocyanin extraction and yield from Spirulinaplatensis. Int J Curr Microbiol Appl Sci 3(8):904–909

Spolaore P, Joannis-Cassan C, Duran E, Isambert A (2006) Commercial applications of microalgae. J Biosci Bioeng 101:87–96

Stanic-Vucinic D, Minic S, Nikolic MR, Velickovic TC (2018) Spirulina Phycobiliproteins as Food Components and Complements. In 1st ed, Jacob-Lopes E, Queiroz Zepka L, Queiroz MI (Eds) Microalgal Biotechnology, London, 129–149

Sun L, Wang S, Chen L, Gong X (2003) Promising fluorescent probes from phycobiliproteins. IEEE J Sel Top Quant 9(2):177–188

Tarko T, Duda-Chodak A, Kobus M (2012) Influence of growth medium composition on synthesis of bioactive compounds and antioxidant properties of selected strains of Arthrospira cyanobacteria. Czech J Food Sci 30:258–267

Tavanandi HA, Mittal R, Chandrasekhar J, Raghavarao KSMS (2018) Simple and efficient method for extraction of C-Phycocyanin from dry biomass of Arthospiraplatensis. Algal Res 31:239–251

Venkataraman LV, Becker EW (1985) Biotechnology & utilization of algae - The Indian experience. Sharada Press, Mangalore, pp 114–115



The authors are grateful to the Principal and Head, Department of Botany, Madras Christian College (Autonomous) for their support and providing the necessary infrastructure. The authors also like to thank DST-FIST for providing the instrument facility.

Author Information

Kathiravan A.
Department of Botany, Madras Christian College (Autonomous) Affiliated to The University of Madras, Chennai, India

Ranjith Kumar R.
Department of Botany, Madras Christian College (Autonomous) Affiliated to The University of Madras, Chennai, India