Egyptian Arthrospira phytosterols: production, identification, antioxidant and antiproliferative activities

  • Gamal S. El BAROTY Cairo University, Faculty of Agriculture, Biochemistry Department, Cairo
  • Hanaa H. Abd El BAKY National Rsearch Center, Plant Biochemistry Department, Dokki, Giza
  • Mahmoud A. SALEH Cairo University, Faculty of Agriculture, Biochemistry Department, Cairo
Keywords: antioxidant; antiproliferative; microalgae Spirulina maxima; phosphorus; phytosterols; sulfur and nitrogen limitation


Cultivation of microalgae as a source of phytosterol and other lipid compounds has gained more attention for commercial applications in pharmaceutical, cosmetic and food function industries. In this research, native Spirulina maxima SM from Egypt was grown in individual cultures containing various concentrations of nitrogen (N), phosphorus (P) and sulfur (S) elements in order to elucidate the effect of its elements on lipid and phytosterols production and to evaluate its antioxidant and antiproliferative activities. The results revealed that the SM was able to grow in different concentrations of testing elements S (from 0.3 to 2.4 mM), P (from 0.3 to 2.4 mM) and N (0.3 to 3.2 mM) with significant differences. A high potential for production of MS biomass, total lipid and phytosterol contents were obtained in individual cultures containing 0.6 mM N, 0.6 mM P and 0.80 mM, respectively. Therefore, these concentrations (combination of S+P+N element) were selected for cultivation of SM at large scale in a column photobioreactor (PBR 300 L) to induce sufficient SM biomass so that, we can obtain an adequate amount of total lipid and phytosterol contents. Phytosterols (PS) of native SM grown in the 300 L PBR were partially purified from unsaponified extracts of SM total lipid followed by its purification by crystallization process. The identification and quantification of PS profile were performed by GC-FID analysis. The results revealed high levels of campsterol, D7-Avena sterol, β-sitosterol, stigmasterol and other compounds. These PS compounds showed marked in vitro superoxide, DPPH and .OH radical scavenging activity, which was comparable with the results obtained with standard antioxidants BHA or α-tocopherol. Moreover, SM phytosterols exhibited anti-proliferative activity against three human cancer cell lines (MCF-7, Hep-G2 and HCT-116) with IC50 values less than 11.62 µg/mL as assessed by in vitro MTT colorimetric method. Thus, SM phytosterol may be considered as a potential natural source of promising ingredient in the future for a range of health applications for human, cosmetic industries and in functional food.


Metrics Loading ...


Abd El Baky HH, El Baz FK, El-Baroty GS (2006). Over-production of lipid rich in linolenic acid by blue green alga Spirulina maxima and its inhibitory effect on carcinoma cells. Advances in Food Sciences 28(4):206-213.

El Baky HH, El Baz FK, El-Baroty GS (2009). Phenolics from Spirulina maxima: over-production and in vitro protective effect of its phenolics on CCl 4 induced hepatotoxicity. Journal of Medicinal Plant Research 3(1):24-30.

Abd El Baky HH, El Baroty GS (2012). Characterization and bioactivity of phycocyanin isolated from Spirulina maxima grown under salt stress. Food & Function 3(4):381-388.

Abd El Baky HH, El-Baroty GS (2013). The potential use of microalgal carotenoids as dietary supplements and natural preservative ingredients. Journal of Aquatic Food Product Technology 22(4):392-406.

Abd El Baky HH, El Baz FK, El Baroty GS, Asker MS, Ibrahim EA (2014). Phospholipids of some marine macroalgae: Identification, antivirus, anticancer and antimicrobial bioactivities. Der Pharma Chemica 6(5):370-382.

Abd El Baky HH, El Baroty GS (2016). Optimization of growth conditions for purification and production of L-asparaginase by Spirulina maxima. Evidence-Based Complementary and Alternative Medicine.

Abd El Baky HH, El Baroty GSA, Ibrahem EA (2016). Functional characters evaluation of biscuits sublimated with pure phycocyanin isolated from Spirulina and Spirulina biomass. Nutricion Hospitalaria 32(1):231-241.

Ahmed F, Zhou W, Schenk PM (2015). Pavlova lutheri is a high-level producer of phytosterols. Algal Research 10:210-217.

Andrade LM, Andrade CJ, Dias M, Nascimento CA, Mendes MA (2018). Chlorella and Spirulina microalgae as sources of functional foods, nutraceuticals, and food supplements; an overview. MOJ Food Processing & Technology 6(1):45-58.

Abdul QA, Choi RJ, Jung HA, Choi JS (2016). Health benefit of fucosterol from marine algae: a review. Journal of the Science of Food and Agriculture 96(6):1856-1866.

Bachiega P, Salgado JM, de Carvalho JE, Ruiz ALT, Schwarz K, Tezotto T, Morzelle MC (2016). Antioxidant and antiproliferative activities in different maturation stages of broccoli (Brassica oleracea Italica) biofortified with selenium. Food Chemistry 190:771-776.

Bai M, Cheng C, Wan H, Lin Y (2011). Microalgal pigments potential as byproducts in lipid production. Journal of the Taiwan Institute of Chemical Engineers 42(5):783-786.

Baskar A, Ignacimuthu S, Paulraj GM, Al Numair KS (2010). Chemopreventive potential of β-Sitosterol in experimental colon cancer model -an in vitro and in vivo study. BMC Complementary and Alternative Medicine 10(1):24.

Bligh EG, Dyer WJ (1959). A rapid method of total extraction and purification. Canadian Journal of Biochemistry and Physiology 37(8):911-917.

Borowitzka MA (2013). High-value products from microalgae-their development and commercialization. Journal of Applied Phycology 25(3):743-756.

Bradford PG, Awad AB (2007). Review: phytosterols as anticancer compounds. Molecular Nutrition and Food Research 51(2):161-170.

Celekli A, Yavuzatmaca M, Bozkurt H (2009). Modeling of biomass production by Spirulina platensis as function of phosphate concentrations and pH regimes. Bioresource Technology 100(14):3625-3629.

Chuanphongpanich S, Nipon T, Duang Buddhasukh D, Pirakitikulr P, Phanichphant S (2006). Stanol synthesis from palm oil distillate. Chiang Mai Journal of Sciences 33(1):109-116.

Chiu SY, Kao CY, Tsai MT, Ong SC, Chen CH, Lin CS (2009). Lipid accumulation and CO2 utilization of Nannochloropsis oculata in response to CO2 aeration. Bioresource Technology 100(2):833-838.

Cilla A, Attanzio A, Barberá R, Tesoriere L, Livrea MA (2015). Anti-proliferative effect of main dietary phytosterols and ß-cryptoxanthin alone or combined in human colon cancer Caco-2 cells through cytosolic Ca+2 and oxidative stressinduced apoptosis. Journal of Functional Foods 12:282-293.

Colla LM, Reinehr CO, Reichert C, Costa JAV (2007). Production of biomass and nutraceutical compounds by Spirulina platensis under different temperature and nitrogen regimes. Bioresource Technolology 98(7):489-1493.

El Baroty GS, El Baz FK, Abd-Elmoein I, Abd El-Baky HH, Ail MM, Ibrahim EA (2011). Evaluation of glycolipids of some Egyptian marine algae as a source of bioactive substances. Electronic Journal of Environmental, Agricultural and Food Chemistry 10(4):2114-21218.

Halliwell B, Gutteridge JMC, Aruoma OI (1987). The deoxyribose method: A simple ‘test tube’ assay for determination of rates constants for reactions of hydroxyl radical. Analytical Biochemistry 165(1):215-224.

Gordon MH, Magos P (1983). The effect of sterols on the oxidation of edible oils. Food Chemistry 10(2):141-147.

Gordillo FJL, Goutx M, Figueroa FL, Niell FX (1998). Effects of light intensity, CO2 and nitrogen supply on lipid class composition of Dunaliella viridis. Journal of Applied Phycology 10(2):135-144.

Jayaprakasha GK, Jadegoud Y, Gowda GA, Patil BS (2010). Bioactive compounds from sour orange inhibit colon cancer cell proliferation and induce cell cycle arrest. Journal of Agricultural and Food Chemistry 58(1):180-186.

Kim Y, Li X, Kang K, Ryu B, Kim S (2014). Stigmasterol isolated from marine microalgae Navicula incerta induces apoptosis in human hepatoma HepG2 cells. BMB Reports 47(8):433-438.

Koh EJ, Kim KJ, Choi J, Kang DH, Lee BY (2018). Spirulina maxima extract prevents cell death through BDNF activation against amyloid beta 1-42 (Aß induced neurotoxicity in PC12 cells. Neuroscience Letters 673:33-38.

Liu ZY, Wang GC, Zhou BC (2008). Effect of iron on growth and lipid accumulation in Chlorella vulgaris. Bioresource Technology 99(11):4717-4722.

Liu M, Sakamaki T, Casimiro MC, Willmarth NE, Quong AA, Ju X (2010). The canonical NF-kappaB pathway governs mammary tumorigenesis in transgenic mice and tumor stem cell expansion. Cancer Research 70(24):10464-10473.

Laverias G, Escola-Gil JC, Lerma E, Julve J, Pons M, Ros E, … Blanco-Vaca F (2013). Phytosterols inhibit the tumor growth and lipoprotein oxidizability induced by a high-fat diet in mice with inherited breast cancer. Journal of Nutritional Biochemistry 24(1):39-48.

Luo X, Su P, Zhang W (2015). Advances in microalgae-derived phytosterols for functional food and pharmaceutical applications. Marine Drugs 13(7):4231-4254.

Mark L, Wells ML, Philippe P, Merchant SS, Camire ME, Craigie JS, … Brawley SH (2016). Algae as nutritional and functional food sources: revisiting our understanding. Journal of Applied Phycology 29(2):949-982.

Martin-Creuzburg D, Von Elert E (2009). Ecological significance of sterols in aquatic food webs. In: Arts MT, Brett MT, Kainz MJ, eds. Lipids in Aquatic Ecosystems. Springer, New York pp 43-64.

McCord JM, Fridovich I (1969). Superoxide dismutase: an enzymic function for erythrocuprein (hemocuprein). Journal of Biological Chemistry 244(22):6049-6055.

Molina Grima E, Belarbi EH, Acien Fernandez FG, Robles Medina A, Chisti Y (2003). Recovery of microalgal biomass and metabolites: process options and economics. Biotechnologies Advances 20(7-8):491-515.

Montserrat-de la Paz S, Fernandez-Arche MA, Bermudez B, Garcia-Gimenez M (2015). The sterols isolated from evening primrose oil inhibit human colon adenocarcinoma cell proliferation and induce cell cycle arrest through upregulation of LXR. Journal of Functional Foods 15:64-69.

Paoletti C, Pushparaj B, Florenzano G, Capella P, Lercker G (1976). Unsaponifiable matter of green and blue‐green algal lipids as a factor of biochemical differentiation of their biomasses: I. Total unsaponifiable and hydrocarbon fraction. Lipids 11(4):266-271.

Piepho M, Martin-Creuzburg DM, Wacker A (2010). Simultaneous effects of light intensity and phosphorus supply on the sterol content of phytoplankton. PLoS ONE 5(12):e15828.

Rao AR, Dayananda C, Sarada R, Shamala TR, Ravishankar GA (2007). Effect of salinity on growth of green alga Botryococcus braunii and its constituents. Bioresource Technology 98(3):560-564.

Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Rice-Evans C (1999). Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radical Biology and Medicine 26(9-10):1231-1237.

Rzamaa A, Dufourc EJ, Arregub B (1994). Sterols from green and blue-green algae grwn on reused waste water. Phytochemistry 37(60):1625-1628.

Scoles DR, Xu X, Wang H, Tran H, Taylor-Harding B, Li A (2010). Liver X receptor agonist inhibits proliferation of ovarian carcinoma cells stimulated by oxidized low density lipoprotein. Gynecologic Oncology 116(1):109-116.

Shanthi G, Premalatha M, Anantharaman N (2018). Effects of L-amino acids as organic nitrogen source on the growth rate, biochemical composition and polyphenol content of Spirulina platensis. Algal Research 35:471-478.

Srigley CT, Haile EA (2015). Quantification of plant sterols/stanols in foods and dietary supplements containing added phytosterols. Journal of Food Composition and Analysis 40:163-176.

Tang D, Han W, Li P, Miao X, Zhong J (2011). CO2 biofixation and fatty acid composition of Scenedesmus obliquus and Chlorella pyrenoidosa in response to different CO2 levels. Bioresource Technology 102(3):3071-3076.

Vivacons M, Moreno JJ (2005). Beta-sitosterol modulates antioxidant enzyme response in RAW 264.7 macrophages. Free Radical Biology and Medicine 39(1):91-97.

Wojcik M, Burzynska-Pedziwiatr I, Wozniak LA (2010). A review of natural and synthetic antioxidants important for health and longetivity. Current Medicinal Chemistry 17(28):3262-3288.

Woyengo TA, Ramprasath VR, Jones PJ (2009). Anticancer effects of phytosterols. European Journal of Clinical Nutrition 63(7):813-820.

Yoshida Y, Niki E (2003). Antioxidant effects of phytosterol and its components. Journal of Nutritional Science and Vitaminology 49(4):277-280.

Yu L, Haley S, Perret J, Harris M, Wilson J, Qian M (2002). Free radical scavenging activity properties of wheat extracts. Journal of Agricultural and Food Chemistry 50(6):1619-1624.

Zhao Y, Chang SK, Qu G, Li T, Cui H (2009). β-Sitosterol inhibits cell growth and induces apoptosis in SGC-7901 human stomach cancer cells. Journal of Agricultural and Food Chemistry 57(12):5211-5218.

Winkler-Moser J (2018). Gas chromatographic analysis of plant sterols. The AOCS Lipid Library.

Zarrouk C (1966). Contribution a letuded unecyanobacterie: influence de divers facteurs physiques et chimiquessur la croissance et la photosynthese de Spirulina maxima (Setchell et Gardner) Geitler[Ph.D. thesis], University of Paris, Paris, France.

How to Cite
El BAROTY, G. S., Abd El BAKY, H. H., & SALEH, M. A. (2020). Egyptian Arthrospira phytosterols: production, identification, antioxidant and antiproliferative activities. Notulae Botanicae Horti Agrobotanici Cluj-Napoca, 48(2), 666-680.
Research Articles