Abstract
Phycocyanin, a blue-coloured pigment, predominantly found and derived from Spirulina sp., has gained researchers' interest due to its vibrant hues and other attractive properties like antioxidant and anti-microbial. However, the lack of reliable and sustainable phycocyanin extraction strategies without compromising the quality has hindered the scaling up of its production processes for commercial purposes. Here in this study, phycocyanin was extracted from wet and dry biomass Spirulina sp., using three different physical cell disruption methods (ultrasonication, homogenization, and freeze–thaw cycles) combined with two different buffers (phosphate buffer and acetate buffer) and water (as control). The result showed that the freeze–thaw method combined with acetate buffer produced the highest yield (25.013 ± 2.572 mg/100 mg) with a purity ratio of 0.806 ± 0.079. Furthermore, when subjected to 30% w/v salt stress, 1.9 times higher phycocyanin yield with a purity ratio of 1.402 ± 0.609 was achieved using the previously optimized extraction method.
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References
Aoki J, Sasaki D, Asayama M (2021) Development of a method for phycocyanin recovery from filamentous cyanobacteria and evaluation of its stability and antioxidant capacity. BMC Biotechnol 21(1):1–10. https://doi.org/10.1186/s12896-021-00692-9
Bachchhav MB, Kulkarni MV, Ingale AG (2017) Enhanced phycocyanin production from spirulina platensis using light emitting diode. J Inst Eng (India): Series E. 98(1):41–45. https://doi.org/10.1007/s40034-016-0090-8
Berns DS, MacColl R (1989) Phycocyanin in physical-chemical studies. Chem Rev 89(4):807–825. https://doi.org/10.1021/cr00094a005
Bhaskar U, Gopalaswamy G, Raghu R (2005) A simple method for efficient extraction and purification of C-phycocyanin from Spirulina platensis Geitler’s. Indian J Exper Biol 43(3):277–279
Chen KH, Wang SSS, Show PL, Lin GT, Chang YK (2018) A rapid and efficient technique for direct extraction of C-phycocyanin from highly turbid Spirulina platensis algae using hydrophobic interaction chromatography in stirred fluidized bed. Biochem Eng J. https://doi.org/10.1016/j.bej.2018.09.005
Chen KH, Wang SSS, Show PL, Hsu SL, Chang YK (2019) Rapid and efficient recovery of C-phycocyanin from highly turbid Spirulina platensis algae using stirred fluidized bed ion exchange chromatography. Sep Purif Technol. 209:636–645. https://doi.org/10.1016/j.seppur.2018.08.057
Chentir I, Doumandji A, Ammar J, Zili F, Jridi M, Markou G, Ben Ouada H (2018) Induced change in Arthrospira sp. (Spirulina) intracellular and extracellular metabolites using multifactor stress combination approach. J Appl Phycol. 30(3):1563–1574. https://doi.org/10.1007/s10811-017-1348-3
Chentir I, Hamdi M, Li S, Doumandji A, Markou G, Nasri M (2018) Stability, bio-functionality and bio-activity of crude phycocyanin from a two-phase cultured Saharian Arthrospira sp. strain. Algal Res 35:395–406. https://doi.org/10.1016/j.algal.2018.09.013
Devi A, Kalwani M, Patil K, Kumari A, Tyagi A, Shukla P, Pabbi S (2023) Microalgal bio-pigments: production and enhancement strategies to enrich microalgae-derived pigments. In: Cyanobacterial Biotechnology in the 21st Century (Issue June). pp. 85–106. https://doi.org/10.1007/978-981-99-0181-4_6
De Morais MG, Da Fontoura Prates D, Moreira JB, Duarte JH, Costa JAV (2018) Phycocyanin from microalgae: Properties, extraction and purification, with some recent applications. Ind Biotechnol 14(1):30–37. https://doi.org/10.1089/ind.2017.0009
Dianursanti Indraputri CM, Taurina Z (2018) Optimization of phycocyanin extraction from microalgae Spirulina platensis by sonication as antioxidant. AIP Conf Proc. https://doi.org/10.1063/1.5023960
Ebrahimi A, Pazuki G, Mozaffarian M, Ahsaie FG, Abedini H (2023) Separation and purification of C-phycocyanin from spirulina platensis using aqueous two-phase systems based on triblock thermosensitive copolymers. Food Bioprocess Technol 16(11):2582–2597. https://doi.org/10.1007/s11947-023-03057-6
El-Naggar NEA, Hussein MH, El-Sawah AA (2017) Bio-fabrication of silver nanoparticles by phycocyanin, characterization, in vitro anticancer activity against breast cancer cell line and in vivo cytotxicity. Sci Rep 7(1):1–20. https://doi.org/10.1038/s41598-017-11121-3
Fernández-Rojas B, Hernández-Juárez J, Pedraza-Chaverri J (2014) Nutraceutical properties of phycocyanin. J Funct Foods 11:375–392. https://doi.org/10.1016/j.jff.2014.10.011
Furuki T, Maeda S, Imajo S, Hiroi T, Amaya T, Hirokawa T, Ito K, Nozawa H (2003) Rapid and selective extraction of phycocyanin from Spirulina platensis with ultrasonic cell disruption. J Appl Phycol 15(4):319–324. https://doi.org/10.1023/A:1025118516888
Glazer AN (1989) Light guides. J Biol Chem 264(1):1–4. https://doi.org/10.1016/s0021-9258(17)31212-7
Gorgich M, Passos MLC, Mata TM, Martins AA, Saraiva MLMFS, Caetano NS (2020) Enhancing extraction and purification of phycocyanin from Arthrospira sp. with lower energy consumption. Energy Rep 6:312–318. https://doi.org/10.1016/j.egyr.2020.11.151
Grosshagauer S, Kraemer K, Somoza V (2020) The true value of spirulina. J Agric Food Chem 68(14):4109–4115. https://doi.org/10.1021/acs.jafc.9b08251
Hsieh-Lo M, Castillo G, Ochoa-Becerra MA, Mojica L (2019) Phycocyanin and phycoerythrin: strategies to improve production yield and chemical stability. Algal Res 42:101600. https://doi.org/10.1016/j.algal.2019.101600
Jaeschke DP, Mercali GD, Marczak LDF, Müller G, Frey W, Gusbeth C (2019) Extraction of valuable compounds from Arthrospira platensis using pulsed electric field treatment. Biores Technol 283(March):207–212. https://doi.org/10.1016/j.biortech.2019.03.035
Jespersen L, Strømdahl LD, Olsen K, Skibsted LH (2005) Heat and light stability of three natural blue colorants for use in confectionery and beverages. Eur Food Res Technol 220(3–4):261–266. https://doi.org/10.1007/s00217-004-1062-7
Kaur M, Saini KC, Ojah H, Sahoo R, Gupta K, Kumar A, Bast F (2022) Abiotic stress in algae: response, signaling and transgenic approaches. J Appl Phycol. 34(4):1843–1869. https://doi.org/10.1007/s10811-022-02746-7
Lauceri R, Cavone C, Chini Zittelli G, Kamburska L, Musazzi S, Torzillo G (2022) High purity grade phycocyanin recovery by decupling cell lysis from the pigment extraction: an innovative approach. Food Bioprocess Technol 16(1):111–121. https://doi.org/10.1007/s11947-022-02926-w
Lee SH, Lee JE, Kim Y, Lee SY (2016) The production of high purity phycocyanin by spirulina platensis using light-emitting diodes based two-stage cultivation. Appl Biochem Biotechnol 178:382–395. https://doi.org/10.1007/s12010-015-1879-5
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. https://doi.org/10.1007/s10337-010-1874-5
Liu C, Li LJ, Wu CY et al (2016) Growth and antioxidant production of Spirulina in different NaCl concentrations. Biotechnol Lett 38:1089–1096. https://doi.org/10.1007/s10529-016-2087-2
Minkova K, Tchorbadjieva M, Tchernov A, Stojanova M, Gigova L, Busheva M (2007) Improved procedure for separation and purification of Arthronema africanum phycobiliproteins. Biotech Lett 29(4):647–651. https://doi.org/10.1007/s10529-006-9274-5
Mishra SK, Shrivastav A, Mishra S (2008) Effect of preservatives for food grade C-PC from Spirulina platensis. Process Biochem 43(4):339–345. https://doi.org/10.1016/j.procbio.2007.12.012
Moraes CC, Sala L, Cerveira GP, Kalil SJ (2011) C-Phycocyanin extraction from Spirulina platensis wet biomass. Braz J Chem Eng 28(1):45–49. https://doi.org/10.1590/S0104-66322011000100006
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. https://doi.org/10.1007/s10811-017-1155-x
Pan-utai W, Iamtham S, Boonbumrung S, Mookdasanit J (2022) Improvement in the sequential extraction of phycobiliproteins from arthrospira platensis using green technologies. Life 12(11):1–19. https://doi.org/10.3390/life12111896
Park WS, Kim HJ, Li M, Lim DH, Kim J, Kwak SS, Kang CM, Ferruzzi MG, Ahn MJ (2018) Two classes of pigments, carotenoids and c-phycocyanin, in spirulina powder and their antioxidant activities. Molecules 23(8):1–11. https://doi.org/10.3390/molecules23082065
Pez Jaeschke D, Rocha Teixeira I, Damasceno Ferreira Marczak L, Domeneghini Mercali G (2021) Phycocyanin from Spirulina: a review of extraction methods and stability. Food Res Int. https://doi.org/10.1016/j.foodres.2021.110314
Pott RWM (2019) The release of the blue biological pigment C-phycocyanin through calcium-aided cytolysis of live Spirulina sp. Color Technol 135(1):17–21. https://doi.org/10.1111/cote.12373
Prabhath GPWA, Shukla SP, Kumar K, Nuwansi KKT (2019) Salinity mediated enhancement in protein and pigment content in Spirulina (Arthrospira) platensis. Indian J Biotechnol 18:323–329
Pradeep HN, Nayak CA (2019) Enhanced stability of C-phycocyanin colorant by extrusion encapsulation. J Food Sci Technol 56(10):4526–4534. https://doi.org/10.1007/s13197-019-03955-8
Purohit A, Kumar V, Chownk M, Yadav SK (2019) Processing-independent extracellular production of high purity C-phycocyanin from spirulina platensis. ACS Biomater Sci Eng 5(7):3237–3245. https://doi.org/10.1021/acsbiomaterials.9b00370
Rajasekaran C, Ajeesh CPM, Balaji S, Shalini M, Siva R, Das R, Fulzele DP, Kalaivani T (2016) Effect of modified Zarrouk’s medium on growth of different spirulina strains. Agric Technol Biol Sci Walailak J Sci Tech. 13(1):67–75
Safari R, Amiri RZ, Esmaeilzadeh Kenari R (2020) Antioxidant and antibacterial activities of C-phycocyanin from common name Spirulina platensis. Iran J Fish Sci. 19(4):1911–1927. https://doi.org/10.22092/ijfs.2019.118129
Sarada R, Pillai MG, Ravishankar GA (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(8):795–801. https://doi.org/10.1016/S0032-9592(98)00153-8
Saran S, Puri N, Dut Jasuja N, Kumar M, Sharma G (2016) Optimization, purification and characterization of phycocyanin from spirulina platensis. Int J Appl Pure Sci Agric 2(3):15–21
Seyed Yagoubi A (2017) The effect of different lipids on physicochemical characteristics and stability of phycocyanin-loaded solid lipid nanoparticles. Iran J Food Sci Technol 14(67):83–93. https://sid.ir/paper/71628/en
Seyed Yagoubi A, Shahidi F, Mohebbi M, Varidi M, Golmohammadzadeh S (2018) Preparation, characterization and evaluation of physicochemical properties of phycocyanin-loaded solid lipid nanoparticles and nanostructured lipid carriers. J Food Measure Charact 12(1):378–385. https://doi.org/10.1007/s11694-017-9650-y
Shaochen G (2016) Extracting phycocyanin from spirulina and hydrothermal liquefaction of its residues to produce bio-crude oil. MSc diss., University of Illinois at Urbana-Champaign
Soni RA, Sudhakar K, Rana RS (2017) Spirulina—from growth to nutritional product: a review. Trends Food Sci Technol 69:157–171. https://doi.org/10.1016/j.tifs.2017.09.010
Sonia S, Ravindran DA (2019) Cultivation of spirulina using low-cost organic medium and preparation of phycocyanin based ice creams phycocyanin extraction and purification view project spirulina platensis cultivation using organic inputs for enhancing the bioactive compounds of the spirulina platensis view project cultivation of spirulina using low-cost organic medium and preparation of phycocyanin based ice creams. Int J Res Anal Rev. https://doi.org/10.1729/Journal.23044
Tomaselli L, Boldrini G, Margheri MC (1997) Physiological behaviour of Arthrospira (Spirulina) maxima during acclimation to changes in irradiance. J Appl Phycol. 9:37–43
WachdaHadiyanto H, Harjanto GD, Huzain ML, Aji RW (2019) Production of antioxidant C-phycocyanin using extraction process of Spirulina platensis in large scale industry. IOP Conf Series Mater Sci Eng. 633(1):1–6. https://doi.org/10.1088/1757-899X/633/1/012025
Wang F, Yu X, Cui Y, Xu L, Huo S, Ding Z, Hu Q, Xie W, Xiao H, Zhang D (2023) Efficient extraction of phycobiliproteins from dry biomass of Spirulina platensis using sodium chloride as extraction enhancer. Food Chem. 406:135005. https://doi.org/10.1016/j.foodchem.2022.135005
Xie Y, Jin Y, Zeng X et al (2015) Fed-batch strategy for enhancing cell growth and C-phycocyanin production of Arthrospira (Spirulina) platensis under phototrophic cultivation. Bioresour Technol 180:281–287. https://doi.org/10.1016/j.biortech.2014.12.073
Yang W, Wang F, Liu LN, Sui N (2020) Responses of membranes and the photosynthetic apparatus to salt stress in cyanobacteria. Front Plant Sci 11(June):1–10. https://doi.org/10.3389/fpls.2020.00713
Yu J (2017) Application of an ultrafine shearing method for the extraction of C-phycocyanin from spirulina platensis. Molecules 22(11):1–11. https://doi.org/10.3390/molecules22112023
Yu C, Hu Y, Zhang Y et al (2024) Concurrent enhancement of biomass production and phycocyanin content in salt-stressed arthrospira platensis: a glycine betaine- supplementation approach. Chemosphere. 353:141387. https://doi.org/10.1016/j.chemosphere.2024.141387
Zhuxin L, Biao Y, Badamkhand D, Yifan C, Honghong S, Xiao X, Mingqian T, Zhixiang W, Chongjiang C (2023) Carboxylated chitosan improved the stability of phycocyanin under acidified conditions. Int J Biol Macromol. 233:123474. https://doi.org/10.1016/j.ijbiomac.2023.123474
Acknowledgements
The authors thank the Department of Biotechnology and Medical Engineering of the National Institute of Technology Rourkela for providing the research facility.
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The research was financially sponsored by Department of Science and Technology, Government of India (DST, GOI) under Entrepreneurship in Residence (EIR) (Fellow letter No.:KIIT-TBI/INIDHI-EIR/10/09).
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Kerthika Devi Athiyappan: conceptualization, methodology, investigation, data curation, writing—original draft; Rayanee Chaudhuri: conceptualization, methodology, investigation, data curation, review & editing; Balasubramanian Paramasivan: review & editing, supervision, final approval.
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Athiyappan, K.D., Chaudhuri, R. & Balasubramanian, P. Enhancing phycocyanin yield from Spirulina sp. under salt stress using various extraction methods. Arch Microbiol 206, 258 (2024). https://doi.org/10.1007/s00203-024-03968-8
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DOI: https://doi.org/10.1007/s00203-024-03968-8