EFFECT OF H2O2 OXIDATION STRESS ON CAROTENOID PRODUCTION IN CHLAMYDOMONAS REINHARDTII AND ITS ANTIOXIDANT ACTIVITY

Öz

The effect of H2O2 oxidation stress on carotenoid production in C. reinhardtii and its antioxidant properties were investigated in this study. For this purpose, the amount of carotenoids determined by HPLC-DAD, total of phenolic contents and antioxidant capacities determined by Folin and CHROMAC methods respectively, in different oxidative stress conditions were studied. When the concentration of H2O2 was increased during the incubation period, total of phenolic content and antioxidant capacity value were decreased. In the same incubation period, HPLC-PDA results showed highest amounts of carotenoids in microalgae exposed to 1 μM H2O2 oxidative stress and it was thought that toxic dose might be in 20 μM oxidation media for microalgae. Thus, when the oxidative stress conditions were changed, the amounts of carotenoids and the structure of carotenoids could be changed. This study is important that the relationship between carotenoid and the power of oxidation stress in microalgae system.

Anahtar Kelimeler

• Antioxidant
• Chlamydomonas reinhardtii
• Ultrasonic-assisted extraction
• HPLC
• Carotenoid
• oxidative stress

H2O2 OKSİDASYON STRESİNİN CHLAMYDOMONAS REINHARDTİİ MİKROALGİNİN KAROTENOİD ÜRETİMİ VE ANTİOKSİDAN AKTİVİTESI ÜZERİNE ETKİSİ

Öz

Bu çalışmada C. reinhardtii mikroalginin karotenoid üretimi ve antioksidan özellikleri üzerine H2O2 molekülünün oksidasyon etkisi incelenmiştir. Bu amaçla karotenoid miktarları HPLC-DAD cihazı ile, toplam fenolik madde ve antioksidan kapasite miktarları sırasıyla Folin ve CHROMAC yöntemleri ile tayin edilmiştir. Oksidatif stress süresi arttıkça karotenoid miktarlarında da artma görülmüştür. İnkübasyon süresi boyunca H2O2 konsantrasyonu arttırıldığında, toplam fenolik madde ve antioksidan kapasite değeri azalmıştır. Aynı inkübasyon süresi boyunca HPLC-DAD sonuçlarına göre, 1 μM H2O2 oksidatif strese maruz kalan mikroalglerde en yüksek miktarda karotenoid sentezlendiği ve mikroalgler için toksik dozun 20 μM oksidasyon ortamında olabileceği anlaşılmıştır. Böylece, oksidatif stres koşulları değiştiğinde, karotenoidlerin miktarları ve karotenoidlerin yapısı değiştirilebilir. Bu çalışma, mikroalg sisteminde, karotenoid sentezi ile oksidasyon stresi arasındaki ilişkinin açıklanması açısından önemlidir.

Anahtar Kelimeler

• Antioksidan
• Chlamydomonas reinhardtii
• Ultrasonik destekli ekstraksiyon
• HPLC
• Karotenoid
• Oksidatif stres

Kaynakça

1. Abd El Baky, H. H., El-Baroty, G. S. (2013). Healthy Benefit of Microalgal Bioactive Substances. Journal of Aquatic Science, 1(1), 11-22. https://doi.org/10.12691/jas-1-1-3.
2. Apel, K., Hirt, H. (2004). Reactive oxygen species: metabolism, oxidative stress, and signal transduction. Annual Review of Plant Biology, 55, 373–399. https://doi.org/10.1146/annurev.arplant.55.031903.141701
3. Butnariu, M. (2016). Methods of analysis (extraction, separation, identification and quantification) of carotenoids from natural products. Journal of Ecosystem & Ecography, 6, 193. https://doi.org/10.4172/2157-7625.1000193
4. Cazzonelli, C. I. (2011). Carotenoids in nature: insights from plants and beyond. Functional Plant Biology, 38, 833–847. https://doi.org/10.1071/FP11192.
5. Cordero, B. F., Couso, I., León, R., Rodríguez, H., Vargas, M. Á. (2011). Enhancement of carotenoids biosynthesis in Chlamydomonas reinhardtii by nuclear transformation using a phytoene synthase gene isolated from Chlorella zofingiensis. Applied Microbiology Biotechnology, 91, 341–351. https://doi.org/10.1007/s00253-011-3262-y.
6. Çakmak, Z. E., Ölmez, T. T., Çakmak, T., Menemen, Y., Tekinay, T. (2015). Antioxidant response of Chlamydomonas reinhardtii grown under different element regimes. Phycological Research, 63, 202–211. https://doi.org/10.1111/pre.12096.
7. El-Bahr, S. M. (2013). Biochemistry of free radicals and oxidative stres. Science International, 1(5), 111-117. https://doi.org/10.17311/sciintl.2013.111.117.
8. Erdoğan, A., Çağır, A., Dalay, M. C., Eroğlu, A. E. (2015). Composition of carotenoids in Scenedesmus protuberans: application of chromatographic and spectroscopic methods. Food Analytical Methods, 8, 1970–1978. https://doi.org/10.1007/s12161-015-0088-8.

9. Goiris, K., Colen, W. V., Wilches, I., León-Tamariz, F., Cooman, L. D., Muylaert, K. (2015). Impact of nutrient stress on antioxidant production in three species of microalgae. Algal Research, 7: 51–57. https://doi.org/10.1016/j.algal.2014.12.002.
10. Guedes, A. C., Amaro, H. M., Malcata, F. X. (2011). Microalgae as sources of carotenoids. Marine Drugs, 9, 625-644. https://doi.org/10.3390/md9040625
11. Güçlü K., Altun M., Özyürek M., Karademir S. E., Apak R. (2006). Antioxidant capacity of fresh, sun- and sulphited-dried Malatya apricot (Prunus armeniaca) assayed by CUPRAC, ABTS/TEAC and folin methods. International Journal of Food Science and Technology, 41, 76–85. https://doi.org/10.1111/j.1365-2621.2006.01347.x
12. Hoham, R.W., Bonome, T.A., Martin, C.W., Leebens-Mack, J.H. (2002). A combined 18S rDNA and rbcL phylogenetic analysis of Chloromonas and Chlamydomonas (Chlorophyceae, Volvocales) emphasizing snow and other cold-temperature habitats. Journal of Phycology, 38,1051–1064. https://doi.org/10.1046/j.1529-8817.2002.t01-1-01227.x
13. Işık, E., Şahin, S., Demir, C. (2013). Development of a new chromium reducing antioxidant capacity (CHROMAC) assay for plants and fruits. Talanta, 111, 119–124. https://doi.org/10.1016/j.talanta.2013.02.053
14. Kessler, E. 1976. Comparative physiology, biochemistry, and taxonomy of Chlorella (Chlorophyceae). Plant Systematics and Evolution, 125, 129–138. https://doi.org/10.1007/BF00986146.
15. Minhas, A. K., Hodgson, P., Barrow, C. J., Adholeya, A. (2016). A review on the assessment of stress conditions for simultaneous production of microalgal lipids and carotenoids. Frontiers in Microbiology, 7, 546. https://doi.org/10.3389/fmicb.2016.00546.
16. Mittler, R. (2002). Oxidative stress antioxidants and stress tolerance. Trends Plant Science, 7, 405–410. https://doi.org/10.1016/s1360-1385(02)02312-9.
17. Mittler, R., Vanderauwera, S., Gollery, M., Van Breusegem, F. (2004). Reactive oxygen gene network of plants. Trends Plant Science, 9, 490–498. https://doi.org/10.1016/j.tplants.2004.08.009
18. Moran, N. A., Jarvik, T. (2010). Lateral transfer of genes from fungi underlies carotenoid production in aphids. Science, 30, 328(5978):624-627. https://doi.org/10.1126/science.1187113
19. Nasır, N. T. B. M., Şahin, S., Çakmak, Z. E., Çakmak, T. (2017). Optimization of ultrasonic-assisted extraction via multiresponse surface for high antioxidant recovery from Chlorella vulgaris (Chlorophyta). Phycologia, 56 , 561–569. https://doi.org/10.2216/16-132.1
20. Neill, S. J., Desikan, R., Clarke, A., Hurst, R.D., Hancock, J.T. (2002). Hydrogen peroxide and nitric oxide as signalling molecules in plants. Journal of Experimental Botany, 53, 1237–1247. https://doi.org/10.1093/jexbot/53.372.1237.
21. Nikulina, K., Chunaev, A.S., Boschetti, A. (2016). Accumulation of zeta-carotene in Chlamydomonas reinhardtii under control of the ac5 nuclear gene. Plant Cell Reports, 19, 37-42. https://doi.org/10.1007/s002990050707.
22. Olasehinde, T. A., Olaniran, A. O., Okoh, A. I. (2017). Therapeutic potentials of microalgae in the treatment of alzheimer’s disease. Molecules, 22, E480. https://doi.org/10.3390/molecules22030480
23. Rao, A. V., Rao, L. G. (2007). Carotenoids and human health. Pharmacological Research, 55, 207–216. https://doi.org/10.1016/j.phrs.2007.01.012
24. Rippka, R., Deruelles, J., Waterbury, J.B., Herdman, M., Stanier, R.Y. (1979). Generic assignments, strain histories and properties of pure cultures of cyanobacteria. Journal of General Microbiology, 111, 1–61. https://doi.org/10.1099/00221287-111-1-1
25. Rochaix, J. D. (1995). Chlamydomonas reinhardtii as the photosynthetic yeast. Annual Review of Genetics, 29, 209-230. https://doi.org/10.1146/annurev.ge.29.120195.001233
26. Siems, W., Wiswedel, I., Salerno, C., Crifo, C., Augustin, W., Schild, L., Langhans, C.D., Sommerburg, O. (2005). β-carotene breakdown products may impair mitochondrialfunctions - potential side effects of high-dose β-carotene supplementation. Journal of Nutritional Biochemistry, 16, 385–397. https://doi.org/10.1016/j.jnutbio.2005.01.009.
27. Şahin, S., Aybastier, Ö., Işık, E. (2013). Optimisation of ultrasonic assisted extraction of antioxidant compounds from Artemisia absinthium using response surface methodology. Food Chemistry, 141, 1361–1368. https://doi.org/10.1016/j.foodchem.2013.04.003
28. Woodall, A.A., Lee, S.W.M., Weesie, R.J., Jackson, M.J., Britton, G., (1997). Oxidation of carotenoids by free radicals: relationship between structure and reactivity. Biochimica et Biophysica Acta, 1336, 33–42.