A comparative study on antioxidant and cytotoxic effects of Oscillatoria amphibia and Spirulina platensis C-phycocyanin and crude extracts

Burcu Tepedelen Erbaykent; İsmet Gurhan-deliloglu; Meltem Conk Dalay

EN TR

A comparative study on antioxidant and cytotoxic effects of Oscillatoria amphibia and Spirulina platensis C-phycocyanin and crude extracts

Abstract

In this study, antioxidant and cytotoxic effects of Oscillatoria amphibia Ege-Macc-14 (Lake Acıgöl-Denizli, Turkey) C-phycocyanin and the crude extract were compared with Spirulina platensis. Since S. platensis is seen as the sole economical source of C-phycocyanin this comparison study may provide novel information for investigating new sources of C-PC and besides new promising cytotoxic compounds. Both species are cultivated under laboratory conditions and both the crude extracts and the C-PCs were obtained. Since O. amphibia has the possibility to produce toxic compounds, in order to figure out the toxicity of the biomass, the aqueous crude extracts of O. amphibia were tested against the brine shrimp directly. The aqueous extracts were found as non-toxic even at the highest concentrations (10 mg.ml-1). Human hepatocellular carcinoma (HEP-3B) cell line was used to investigate the cytotoxic effect of the extracts. The determined IC50 values of S. platensis C-PC, O. amphibia C-PC were 13.27 μg/ml, 4.19 μg/ml respectively and for S. platensis crude extract and O. amphibia crude extract the values were 16.34 μg/ml and 12.94 μg/ml, respectively. The potential antioxidant activity of the crude extracts and the pure C-PCs were evaluated by DPPH free radical, hydroxyl radical scavenging activity assays. As a result, although the pure C-PC of both cyanobacteria showed cytotoxic and antioxidant activities, the highest activity levels were obtained from O. amphibia extracts.

Keywords

Oscillatoria amphibia ve Spirulina platensis’in C-Fikosiyanin ve ham ekstraktlarının antioksidan ve sitotoksik etkileri üzerine karşılaştırmalı çalışma

Öz

Bu çalışmada Oscillatoria amphibia (Ege Macc-14, Acıgöl-Denizli, Türkiye) anti oksidan ve sitotoksik etkileri, Spirulina platensis’in ham ekstrakt ve fikosiyanini ile karşılaştırılmıştır. S. Platensis’in tek ticari C-fikosiyanin kaynağı olarak görülmesinden dolayı bu çalışma yeni fikosiyanin kaynakları araştırma ve umut vaat eden sitoksik bileşik araştırmalarında yeni veriler sağlayabilir. Her iki tür de laboratuar koşulları altında kültüre alınarak ham ekstrakt ve C-fikosiyanin ekstraksiyonları yapılmıştır. O. amphibia’nın sitokisik bileşikler üretebilme potansiyelinden dolayı biyomasın toksisitesini araştırmak üzere sulu O. amphibia ekstreleri hazırlanmış ve artemialara direkt olarak uygulanmıştır. Sulu ham ekstraktlar en yüksek konsantrasyonda (10 mg.ml-1) bile toksik bulunmamıştır. Ekstraktların sitotoksisitesini belirlemek amacıyla insan karaciğer karsinoma (HEP-3B) hücre hattı kullanılmıştır. S. platensis C-fikosiyanini, O. amphibia Cfikosiyaninleri için IC50 değerleri sırasıyla 13,27 μg/ml ve 4,19 μg/ml olarak hesaplanmış ve bu değerler ham ekstratlar içinde yine sırasıyla 16,34 μg/ml ve 12,94 μg/ml olarak bulunmuştur. Ham ekstrakt ve C-fikosiyaninlerin antioksidan potansiyelleri ise DPPH serbest radikal ve hidroksil süpürme aktivite testleriyle belirlenmiştir. Sonuç olarak her iki siyanobakterinin saf C-fikosiyaninleri sitotoksik ve antioksidan aktivite göstermeleriyle birlikte en yüksek değerler O. amphibia ekstraktları için bulunmuştur

Anahtar Kelimeler

References

Anamika, P., Sandhya, M., Richa, P. Ghosh, P.K., 2005. Purification and characterization of C-Phycocyanin from cyanobacterial species of marine and freshwater habitat. Protein Expression and Purify, 40(2):248-255. doi:10.1016/j.pep.2004.10.028
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 Sciences, 75(19):2353-2362. doi:10.1016/j.lfs.2004.06.004
Boussiba, S., Richmond, A., 1979. Isolation and characterization of phycocyanins from the blue-green alga Spirulina platensis. Archives of Microbiology, 120:155-159. doi:10.1007/BF00409102
Callegari, J.P., 1989. Feu Vert Pour Les Microalgues. Biofuture, 76:25-40.
Conk-Dalay, M., Hatipoğlu, S., Sağlam, H., 2005. Phycocyanine content and toxicity of Oscillatoria amphibia agardh isolated from Acı Lake, Turkey. Tropical and Subtropical Cyanoprokaryota Workshop. Gran Canaria, Las Palmas, Spain.
Dubois, M., Gilles, K.A., Hamilton, J.K., Rebers, P.S., Smith, F., 1956. Colorimetric method for determination of sugars and related substances. Analytical Chemistry, 28:350–356. doi:10.1021/ac60111a017
Gustafson, K.R., Cardellina II, J.H., Fuller, R.W., Weislow, O.S., Kiser, R.F. Snader, K.M., Patterson, G.M.L., Boyd, M.R., 1989. AIDS-Antiviral sulfolipids from cyanobacteria (blue-green algae). Journal of the National Cancer Institute, 81(16):1254-1258. doi:10.1093/jnci/81.16.1254
Ganapathi, P., Chethana, S., Sridevi, A.S., Raghavarao, K.S.M.S., 2006. Method to obtain C-phycocyanin of high purity. Journal of Chromatography A, 1127(-2):76–81. doi:10.1016/j.chroma.2006.05.073

Glazer, A.N., 1994. Phycobiliproteins-a family of valuable, widely used fluorophores. Journal of Applied Physiology, 6:105-112.
Gonzalez, R., Rodriguez, S., Romay, C., Ancheta, O., Gonzalez, A., Armesto, J., Remirez, D., Merino, N., 1999. Anti-inflammatory activity of phycocyanin extract in acetic acid-induced colitis in rats. Pharmacological Research, 39(1):55–59. doi:10.1006/phrs.1998.0409
Grossman, A.R., Schaefer, M.R., Chiang, G.G., Collier, J.L., 1993. The phycobilisome, a light-harvesting complex responsive to environmental conditions. Microbiology Reviews, 5:725–749.
Hayashi, O., Katoh, T., 1994. Enhancement of antibody production in mice by dietary Spirulina platensis. Journal of Nutritional Science and Vitaminology. 40:431-441. doi:10.3177/jnsv.40.431
Hsiao, G., Chou, P.H., Shen, M.Y., Chou, D.S., Lin, C.H., Sheu, J.R., 2005. C-Phycocyanin, a Very Potent and Novel Platelet Aggregation Inhibitor from Spirulina platensis. Journal of Agricalturel and Food Chemistry, 53:7734-7740. doi:10.1021/jf051352y
Jaouen, P., Lepine, B., Rossignol, N., Royer, R., Quéméneur, F., 1999. Clarification and concentration with membrane technology of a phycocyanin solution extracted from Spirulina platensis. Biotechnology Techniques, 13:877-881. doi:10.1023/A:1008980424219
Komarek, J., 2010. Recent changes (2008) in cyanobacteria taxonomy based on a combination of molecular background with phenotype and ecological consequences (genus and species concept). Hydrobiologia, 639:245-259. doi:10.1007/s10750-009-0031-3
Laughlin, J.L., 1991. Assay for. Bioactivity, In: Methods in Plant Biochemistry, K. Hostetman (Ed.), Academic Press, London. Vol. 6:1-31.
Li, Y., Zhang, J.P., Xie, J., Zhao, J.Q., Jiang, L.J., 2001. Temperature induced decoupling of phycobilisomes from reaction centers. Biochimica et Biophysica Acta, 1504:229-234. doi:10.1016/S0005-2728(00)00250-4
Liu, Y., Xu, L., Cheng, N., Lin, L., Zhang, C., 2000. Inhibitory effect of phycocyanin from Spirulina platensis on the growth of human leukemia K562 cells. Journal of Applied Phycology, 12:125-130. doi:10.1023/A:1008132210772
Lorenz, R.T., 1998. Quantitative Analysis of C-Phycocyanin from Spirulina pacifica (Low Temperature Method). Cyanotech Corporation, Spirulina Pacifica Technical Bulletin #004.
Lowry, O.H., Rosebrough, N.J., Farr, A.L., Randall, R.J., 1951. Protein measurement with the Folin-Phenol reagents. Journal of Biological Chemistry, 193:265-275.
Madhyastha, H.K., Radha, K.S., Sugiki, M., Omura, S., Maruyama, M., 2006. Purification of c-phycocyanin from Spirulina fusiformis and its effect on the induction of urokinase-type plasminogen activator from calf pulmonary endothelial cells. Phytomedicine, 13:564-569. doi:10.1016/j.phymed.2005.09.006
Meyer, B.N., Ferrigni, N.R., Putnam, J.E., Jacobsen, L.B., Nichols, D.E., McLaughlin, J.L., 1982. Brine shrimp: a convenient general bioassay for active plant constituents. Planta Medica, 45:31-34. doi:10.1055/s-2007-971236
Minkova, K.M., Tchernov, A.A., Tchorbadjieva, M.I., Fournadjieva, S.T., Antova, R.E., Busheva, M.Ch., 2003. Purification of C-phycocyanin from Spirulina (Arthrospira) fusiformis. Journal of Biotechnology, 102:55-59. doi:10.1016/S0168-1656(03)00004-X
Mosmann, T., 1983. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. Journal of Immunological Methods, 65(1-2):55-63. doi:10.1016/0022-1759(83)90303-4
Okadjima, T., Tanabe, T., Yasuda, T., 1993. Nonurea sodium dodecyl sulfate- polyacrylamide gel electrophoresis with high-molarity buffers for the separation of protein and peptides. Analytical Biochemistry, 211:293- 300. doi:10.1006/abio.1993.1272
Pardhasaradhi, B.V.V., Ali, A.A.M., Kumari, A.L., Reddanna, P., Khar, R., 2003. Phycocyanin mediated apoptosis in AK-5 tumor cells involves down-regulation of Bcl-2 and generation of ROS. Moleculer Cancer Therapeutic, 2:1165-1170.
Qureshi, M., Ali, RA., 1996a. Phagocytic potential of Feline macrophages after exposure to a water soluble extract of Spirulina in vitro. Immunopharmacol Immunotoxicolology, 18(3):342-354.
Qureshi, M., Ali, RA., 1996b. Spirulina platensis exposure enhances macrophage phagocytic function in cats. Immunopharmacol Immunotoxicolology, 18(3):457-463. doi:10.3109/08923979609052747
Reddy, M.C., Subhashin, J., Mahipal, S.V.K., Bhat, V.B., Reddy, S.P., Kiranmai, G., Madyastha, K.M., Reddanna, P., 2003. C-Phycocyanin, a selective cyclooxygenase-2 inhibitor, induces apoptosis in lipopolysaccharide-stimulated RAW 264.7 macrophages. Biochemical and Biophysical Research Communications, 304:385-392. doi:10.1016/S0006-291X(03)00586-2
Reis, A., Mendes, A., Lobo-Fernandes, H., Empis, J.A., Novais, J.M., 1998. Production, extraction and purification of phycobiliproteins from Nostoc sp. Bioresource Technology, 66:181-187. doi:10.1016/S0960-8524(98)00064-9
Romay, C., Armesto, J., Remirez, D., Gonzalez, R., Ledon, N., Garcia, I., 1998. Antioxidant and anti-inflammatory properties of C-phycocyanin from blue-green algae. Inflammation Research, 47:36–41. doi:10.1007/s000110050256
Romay, C., Gonzalez, R., 2000. Phycocyanin is an antioxidant protector of human erythrocytes against by peroxyl radicals. Journal of Pharmacy and Pharmacology, 52:8-367. doi:10.1211/0022357001774093
Romay C., González, R., Ledón, N., Ramirez, D., Rimbau, V., 2003. C- Phycocyanin: a biliprotein with antioxidant, anti-inflammatory and neuroprotective effects. Current Protein and Peptide Science, 4:207-16. doi:10.2174/1389203033487216
Schreiber, E., 1927. Die Reinkultur von marinen phytoplankton und deren bedeutung für die erforschung der produktionsfähigkeit des meerwassers. Wiss Meeresuntersuch N.F., 10:1-34.
Singh, R., Singh, S., Kumar, S., Arora, S., 2007. Free radical-scavenging activity of acetone extract/fractions of Acacia auriculiformis A. Cunn. Food Chemistry, 103:1403-1410. doi:10.1016/j.foodchem.2006.10.056
Sreemantula, S., Nammi, S., Kolanukonda, R., Koppula, S., Boini, K.M., 2005. Adaptogenic and nootropic activities of aqueous extract of Vitis vinifera (grape seed): an experimental study in rat model. BMC Complementary and Alternative Medicine, 5(1):1-19. doi:10.1186/1472-6882-5-1
Stocker, R., Mcdonagh, A.F., Glazer, A.N., Ames, B.N. 1990. Antioxidant activities of bile pigments: biliverdin and bilirubin. Methods in Enzymology, 186-301.
Subhashini, J., Mahipal, S.V., Reddy, M.C., Mallikarjuna, R.M., Rachamallu, A., Reddanna, P., 2004. Molecular mechanisms in C-Phycocyanin induced apoptosis in human chronic myeloid leukemia cell line-K562. Biochemical Pharmacology, 68:453-462. doi:10.1016/j.bcp.2004.02.025
Surakka, A., Sihvonen, L.M., Lehtimäki, J.M., Wahlsten, M., Vuorela, P., Sivonen, K., 2005. Benthic cyanobacteria from the Baltic Sea contain cytotoxic Anabaena, Nodularia and Nostoc strains and an apoptosis- inducing Phormidium strain. Environmental Toxicology, 20:285-292. doi:10.1002/tox.20119
Velioğlu, Y.S., Mazza, G., Gao, L., Oomah, B.D., 1998. Antioxidant activity and total phenolics in selected fruits, vegetables and grain products. Journal of Agricaltural and Food Chemistry, 46:375-379. doi:10.1021/jf9801973
Xu, J., Chen, S., Hu, Q., 2005. Antioxidant activity of brown pigment and extracts from black sesame seed (Sesamum indicum L). Food Chemistry, 91(1):79-83. doi:10.1016/j.foodchem.2004.05.051
Yi-Ming, Z., Fheng, C., 1999. A simple method for efficient separation and purification of C-Phycocyanin and Allophycocyanin from Spirulina platensis. Biotechnology Techniques, 13:601-603. doi:10.1023/A:1008914405302
Yoshida, A., Takagaki, Y., Nishimune, T., 1996. Enzyme immunoassay for phycocyanin as the main component of Spirulina color in foods. Bioscience Biotechnology and Biochemistry, 60:57-60. doi:10.1271/bbb.60.57
Zarrouk, C., 1966. Contribution A L’etude du Cyanophyceae. Influence de Divers Facteurs Physiques et Chimiques Sur La Croissance et La Photosynthese de Spirulina Maxima (Setch Et Gardner) Geitl., Phd, Paris.

Details

Primary Language

English

Subjects

-

Journal Section

Note

Authors

Burcu Tepedelen Erbaykent

İsmet Gurhan-deliloglu

Meltem Conk Dalay

Publication Date

March 1, 2012

Submission Date

December 7, 2015

Acceptance Date

-

Published in Issue

Year 2012 Volume: 29 Number: 1

IZ

https://izlik.org/JA68WG73HC

Cite

RIS / Bibtex

APA

Erbaykent, B. T., Gurhan-deliloglu, İ., & Dalay, M. C. (2012). A comparative study on antioxidant and cytotoxic effects of Oscillatoria amphibia and Spirulina platensis C-phycocyanin and crude extracts. Ege Journal of Fisheries and Aquatic Sciences, 29(1), 1-7. https://izlik.org/JA68WG73HC