Exploring of the Green Algae Chlorella Vulgaris Potential for Phenantherene Biodegredation
Abstract
The distribution of Polycyclic Aromatic Hydrocarbons (PAHs) as a group of toxic, and persistent aromatic pollutants in the environment is rapidly enhancing. These compounds have adverse impacts on the health of living organisms. It is necessary to find an effective method for the elimination of PAHs from the ecosystems. In the last decades, phycoremediation as an effective, low-cost and ecofriendly technology for the cleanup of different pollutants has gained a great attention. Hence, the present study has been focused on potential of the green microalga Chlorella vulgaris for degradation of phenanthrene as a toxic 3-ring PAHs. The impact of the phenanthrene on Chlorella vulgaris cells was evaluated by assay of alga growth, protein assay and Gas Chromatography–Mass Spectrometry (GC–MS) analysis. Four different concentrations (2, 10, 25 and 50 mg L-1) of phenanthrene were selected for the study. Intriguingly, optical density, as a growth factor of algae, was enhanced when treated with 2 mg L-1 of phenanthrene after 7 days exposure in comparison to control. The cellular growth and total protein content were decreased as the concentration of phenanthrene increased from 10 to 50 mg L-1 at the same exposure. Furthermore, GC/MS technique explained the biological degradation of phenanthrene in the present research and accordingly, a number of intermediate by-products were identified. The obtained results confirmed that phenanthrene is able to induce cytotoxicity in Chlorella cells in high concentrations and subsequently Chlorella vulgaris has noticeable potential in its biodegradation.
Keywords:
Biodegradation, Chlorella vulgaris, Phenanthrene, Green microalgae, PhycoremediationReferences
- [1] Hong, Y. W., Yuan, D. X., Lin, Q. M., & Yang, T. L. (2008). Accumulation and biodegradation of phenanthrene and fluoranthene by the algae enriched from a mangrove aquatic ecosystem. Marine pollution bulletin, 56(8), 1400-1405. https://doi.org/10.1016/j.marpolbul.2008.05.003
- [2] Jajoo, A., Mekala, N. R., Tomar, R. S., Grieco, M., Tikkanen, M., & Aro, E. M. (2014). Inhibitory effects of polycyclic aromatic hydrocarbons (PAHs) on photosynthetic performance are not related to their aromaticity. Journal of photochemistry and photobiology b: Biology, 137, 151-155. https://doi.org/10.1016/j.jphotobiol.2014.03.011
- [3] Patel, J. G., Kumar, J. I. N., Kumar, R. N., & Khan, S. R. (2016). Biodegradation capability and enzymatic variation of potentially hazardous polycyclic aromatic Hydrocarbons—Anthracene and Pyrene by Anabaena fertilissima. Polycyclic aromatic compounds, 36(1), 72–87. https://doi.org/10.1080/10406638.2015.1039656
- [4] Subashchandrabose, S. R., Logeshwaran, P., Venkateswarlu, K., Naidu, R., & Megharaj, M. (2017). Pyrene degradation by Chlorella sp. MM3 in liquid medium and soil slurry: Possible role of dihydrolipoamide acetyltransferase in Pyrene biodegradation. Algal research, 23, 223–232. https://doi.org/10.1016/j.algal.2017.02.010
- [5] Li, F., Zeng, X., Yang, J., Zhou, K., Zan, Q., Lei, A., & Tam, N. F. Y. (2014). Contamination of polycyclic aromatic hydrocarbons (PAHs) in surface sediments and plants of mangrove swamps in Shenzhen, China. Marine pollution bulletin, 85(2), 590–596. https://doi.org/10.1016/j.marpolbul.2014.02.025
- [6] Smith, M. J., Flowers, T. H., Duncan, H. J., & Alder, J. (2006). Effects of polycyclic aromatic hydrocarbons on germination and subsequent growth of grasses and legumes in freshly contaminated soil and soil with aged PAH residues. Environmental pollution, 141(3), 519–525. https://doi.org/10.1016/j.envpol.2005.08.061
- [7] Lei, A. P., Hu, Z. L., Wong, Y. S., & Tam, N. F. Y. (2007). Removal of fluoranthene and Pyrene by different microalgal species. Bioresource technology, 98(2), 273-280. https://doi.org/10.1016/j.biortech.2006.01.012
- [8] Haritash, A. K., & Kaushik, C. P. (2009). Biodegradation aspects of polycyclic aromatic hydrocarbons (PAHs): A review. Journal of hazardous materials, 169(1–3), 1–15. https://doi.org/10.1016/j.jhazmat.2009.03.137
- [9] Zhang, D., Ran, C., Yang, Y., & Ran, Y. (2013). Biosorption of phenanthrene by pure algae and field-collected planktons and their fractions. Chemosphere, 93(1), 61–68. https://doi.org/10.1016/j.chemosphere.2013.04.068
- [10] Asghari, S., Movafeghi, A., Lisar, S. Y. S., Barar, J., & Omidi, Y. (2018). Effects of phenanthrene on growth parameters and antioxidant systems in the green microalga Chlorella Vulgaris. Biointerface res appl chem, 8(8), 3405-3411. https://doi.org/10.33263/BRIAC85.405411
- [11] Patel, M. S., & Tiwari, K. K. (2015). Fluoranthene and acenaphthene metabolism by Chlorella Vulgaris: Identity of intermediates formed during degradation and its growth effect. International journal of recent research and review, 8(1), 26–33. https://www.ijrrr.com/papers8-1/paper4-Fluoranthene%20and%20Acenaphthene%20Metabolism%20by%20Chlorella%20vulgaris%20Identity%20of%20Intermediates%20Formed%20During%20Degradation%20and%20Its%20Growth%20Effect.pdf
- [12] García de Llasera, M. P., Olmos-Espejel, J. D. J., Díaz-Flores, G., & Montaño-Montiel, A. (2016). Biodegradation of benzo (a) Pyrene by two freshwater microalgae Selenastrum capricornutum and Scenedesmus acutus: A comparative study useful for bioremediation. Environmental science and pollution research, 23(4), 3365-3375. https://doi.org/10.1007/s11356-015-5576-2
- [13] Si-Zhong, Y., Hui-Jun, J. I. N., Zhi, W. E. I., Rui-Xia, H. E., Yan-Jun, J. I., Xiu-Mei, L. I., & Shao-Peng, Y. U. (2009). Bioremediation of oil spills in cold environments: A review. Pedosphere, 19(3), 371-381.https://doi.org/10.1016/S1002-0160(09)60128-4
- [14] Rawat, I., Kumar, R. R., Mutanda, T., & Bux, F. (2011). Dual role of microalgae: Phycoremediation of domestic wastewater and biomass production for sustainable biofuels production. Applied energy, 88(10), 3411–3424. https://doi.org/10.1016/j.apenergy.2010.11.025
- [15] Houshani, M., Salehi-Lisar, S. Y., Motafakkerazad, R., & Movafeghi, A. (2019). Uptake and distribution of phenanthrene and Pyrene in roots and shoots of maize (Zea Mays L.). Environmental science and pollution research, 26(10), 9938–9944. https://doi.org/10.1007/s11356-019-04371-3
- [16] Gomes, P. I. A., & Asaeda, T. (2009). Phycoremediation of chromium (VI) by Nitella and impact of calcium encrustation. Journal of hazardous materials, 166(2–3), 1332–1338. https://doi.org/10.1016/j.jhazmat.2008.12.055
- [17] Kalhor, A. X., Movafeghi, A., Mohammadi-Nassab, A. D., Abedi, E., & Bahrami, A. (2017). Potential of the green alga Chlorella Vulgaris for biodegradation of crude oil hydrocarbons. Marine pollution bulletin, 123(1–2), 286–290. https://doi.org/10.1016/j.marpolbul.2017.08.045
- [18] Nazari, F., Movafeghi, A., Jafarirad, S., Kosari-Nasab, M., & Divband, B. (2018). Synthesis of reduced graphene oxide-silver nanocomposites and assessing their toxicity on the green microalga Chlorella Vulgaris. Bionanoscience, 8(4), 997–1007. https://doi.org/10.1007/s12668-018-0561-0
- [19] Peng, S., Zhou, Q., Cai, Z., & Zhang, Z. (2009). Phytoremediation of petroleum contaminated soils by Mirabilis Jalapa L. in a greenhouse plot experiment. Journal of hazardous materials, 168(2–3), 1490–1496. https://doi.org/10.1016/j.jhazmat.2009.03.036
- [20] Huang, G., Chen, F., Wei, D., Zhang, X., & Chen, G. (2018). Biodiesel production by microalgal biotechnology. In Renewable energy (pp. 378–395). Routledge. https://doi.org/10.4324/9781315793245-106
- [21] Kalhor, A. X., Nassab, A. D. M., Abedi, E., Bahrami, A., & Movafeghi, A. (2016). Biodiesel production in crude oil contaminated environment using Chlorella Vulgaris. Bioresource technology, 222, 190–194. https://doi.org/10.1016/j.biortech.2016.09.110
- [22] Fazelian, N., Movafeghi, A., Yousefzadi, M., & Rahimzadeh, M. (2019). Cytotoxic impacts of CuO nanoparticles on the marine microalga Nannochloropsis oculata. Environmental science and pollution research, 26(17), 17499–17511. https://doi.org/10.1007/s11356-019-05130-0
- [23] Prajapati, S. K., Kaushik, P., Malik, A., & Vijay, V. K. (2013). Phycoremediation and biogas potential of native algal isolates from soil and wastewater. Bioresource technology, 135, 232–238. https://doi.org/10.1016/j.biortech.2012.08.069
- [24] Choudhary, P., Prajapati, S. K., & Malik, A. (2016). Screening native microalgal consortia for biomass production and nutrient removal from rural wastewaters for bioenergy applications. Ecological engineering, 91, 221–230. https://doi.org/10.1016/j.ecoleng.2015.11.056
- [25] Upadhyay, A. K., Singh, N. K., Singh, R., & Rai, U. N. (2016). Amelioration of arsenic toxicity in rice: Comparative effect of inoculation of Chlorella Vulgaris and Nannochloropsis sp. on growth, biochemical changes and arsenic uptake. Ecotoxicology and environmental safety, 124, 68–73. https://doi.org/10.1016/j.ecoenv.2015.10.002
- [26] Semple, K. T., Cain, R. B., & Schmidt, S. (1999). Biodegradation of aromatic compounds by microalgae. FEMS microbiology letters, 170(2), 291–300. https://doi.org/10.1111/j.1574-6968.1999.tb13386.x
- [27] Qian, H., Li, J., Pan, X., Sun, L., Lu, T., Ran, H., & Fu, Z. (2011). Combined effect of copper and cadmium on heavy metal ion bioaccumulation and antioxidant enzymes induction in Chlorella Vulgaris. Bulletin of environmental contamination and toxicology, 87(5), 512–516. https://doi.org/10.1007/s00128-011-0365-1
- [28] Suman, T. Y., Rajasree, S. R. R., & Kirubagaran, R. (2015). Evaluation of zinc oxide nanoparticles toxicity on marine algae Chlorella Vulgaris through flow cytometric, cytotoxicity and oxidative stress analysis. Ecotoxicology and environmental safety, 113, 23–30. https://doi.org/10.1016/j.ecoenv.2014.11.015
- [29] Safi, C., Zebib, B., Merah, O., Pontalier, P. Y., & Vaca-Garcia, C. (2014). Morphology, composition, production, processing and applications of Chlorella Vulgaris: A review. Renewable and sustainable energy reviews, 35, 265–278. https://doi.org/10.1016/j.rser.2014.04.007
- [30] El-Sheekh, M. M., Hamouda, R. A., & Nizam, A. A. (2013). Biodegradation of crude oil by Scenedesmus obliquus and Chlorella Vulgaris growing under heterotrophic conditions. International biodeterioration & biodegradation, 82, 67–72. https://doi.org/10.1016/j.ibiod.2012.12.015
- [31] Asghari, S., Rajabi, F., Tarrahi, R., Salehi-Lisar, S. Y., Asnaashari, S., Omidi, Y., & Movafeghi, A. (2020). Potential of the green microalga Chlorella Vulgaris to fight against fluorene contamination: Evaluation of antioxidant systems and identification of intermediate biodegradation compounds. Journal of applied phycology, 32(1), 411–419. https://doi.org/10.1007/s10811-019-01921-7
- [32] Mastral, A. M., Callén, M. S., López, J. M., Murillo, R., García, T., & Navarro, M. V. (2003). Critical review on atmospheric PAH: Assessment of reported data in the Mediterranean basin. Fuel processing technology, 80(2), 183–193. https://doi.org/10.1016/S0378-3820(02)00249-7
- [33] Zhang, J., Cai, L., Yuan, D., & Chen, M. (2004). Distribution and sources of polynuclear aromatic hydrocarbons in mangrove surficial sediments of Deep Bay, China. Marine pollution bulletin, 49(5–6), 479–486. https://doi.org/10.1016/j.marpolbul.2004.02.030
- [34] Wetherell, D. F. (1961). Culture of fresh water algae in enriched natural sea water. Physiologia plantarum, 14(1). https://doi.org/10.1111/j.1399-3054.1961.tb08131.x
- [35] Bradford, M. M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical biochemistry, 72(1–2), 248–254. https://doi.org/10.1016/0003-2697(76)90527-3
- [36] Gong, N., Shao, K., Feng, W., Lin, Z., Liang, C., & Sun, Y. (2011). Biotoxicity of nickel oxide nanoparticles and bioremediation by microalgae Chlorella Vulgaris. Chemosphere, 83(4), 510–516. https://doi.org/10.1016/j.chemosphere.2010.12.059
- [37] Oukarroum, A., Bras, S., Perreault, F., & Popovic, R. (2012). Inhibitory effects of silver nanoparticles in two green algae, Chlorella Vulgaris and Dunaliella tertiolecta. Ecotoxicology and environmental safety, 78, 80–85. https://doi.org/10.1016/j.ecoenv.2011.11.012
- [38] Zhou, H., Wang, X., Zhou, Y., Yao, H., & Ahmad, F. (2014). Evaluation of the toxicity of ZnO nanoparticles to Chlorella Vulgaris by use of the chiral perturbation approach. Analytical and bioanalytical chemistry, 406(15), 3689–3695. https://doi.org/10.1007/s00216-014-7773-0
- [39] Mahjouri, S., Kosari-Nasab, M., Kazemi, E. M., Divband, B., & Movafeghi, A. (2020). Effect of Ag-doping on cytotoxicity of SnO₂ nanoparticles in tobacco cell cultures. Journal of hazardous materials, 381, 121012. https://doi.org/10.1016/j.jhazmat.2019.121012
- [40] Lin, W., Zhang, Z., Chen, Y., Zhang, Q., Ke, M., Lu, T., & Qian, H. (2023). The mechanism of different cyanobacterial responses to glyphosate. Journal of environmental sciences, 125, 258–265. https://doi.org/10.1016/j.jes.2021.11.039
- [41] Jamal, F., Pandey, P. K., & Qidwai, T. (2010). Potential of peroxidase enzyme from Trichosanthes dioica to mediate disperse dye decolorization in conjunction with redox mediators. Journal of molecular catalysis b: Enzymatic, 66(1–2), 177–181. https://doi.org/10.1016/j.molcatb.2010.05.005
- [42] Vafaei, F., Movafeghi, A., Khataee, A. R., Zarei, M., & Lisar, S. Y. S. (2013). Potential of Hydrocotyle Vulgaris for phytoremediation of a textile dye: Inducing antioxidant response in roots and leaves. Ecotoxicology and environmental safety, 93, 128–134. https://doi.org/10.1016/j.ecoenv.2013.03.035
- [43] Bumpus, J. A. (1989). Biodegradation of polycyclic hydrocarbons by Phanerochaete chrysosporium. Applied and environmental microbiology, 55(1), 154–158. https://doi.org/10.1128/AEM.55.1.154-158.1989
- [44] Kim, T. J., Lee, E. Y., Kim, Y. J., Cho, K. S., & Ryu, H. W. (2003). Degradation of Polyaromatic hydrocarbons by Burkholderia Cepacia 2A-12. World journal of microbiology and biotechnology, 19(4), 411–417. https://doi.org/10.1023/A:1023998719787
- [45] Kotterman, M. J. J., Vis, E. H., & Field, J. A. (1998). Successive mineralization and detoxification of benzo [a] Pyrene by the white rot fungus Bjerkandera sp. strain BOS55 and indigenous microflora. Applied and environmental microbiology, 64(8), 2853–2858. https://doi.org/10.1128/AEM.64.8.2853-2858.1998
- [46] Kim, Y.-H., Freeman, J. P., Moody, J. D., Engesser, K.H., & Cerniglia, C. E. (2005). Effects of pH on the degradation of phenanthrene and Pyrene by Mycobacterium vanbaalenii PYR-1. Applied microbiology and biotechnology, 67(2), 275–285. https://doi.org/10.1007/s00253-004-1796-y
- [47] Seo, J. S., Keum, Y. S., & Li, Q. X. (2009). Bacterial degradation of aromatic compounds. International journal of environmental research and public health, 6(1), 278–309. https://doi.org/10.3390/ijerph6010278